Phytomedicine - Articles in Press

Dendrobium candidum extract increases the expression of aquaporin-5 in labial glands from patients with Sjögren's syndrome - Corrected Proof

2010-07-26

Abstract: This study aimed to investigate the mechanism of Dendrobium candidum extract in promoting expression of aquaporin-5 for treatment of Sjögren's syndrome (SS). Sixteen patients with SS suffered from deficient secretion of saliva due to an autoimmune destruction of salivary glands leading to dry mouth symptoms (xerostomia). However, glandular dysfunction also occurred without destruction. Based upon its abnormal distribution in SS salivary glands, a potential role of the water channel protein aquaporin-5 (AQP-5) in the pathogenesis of SS was proposed. After oral administration of D. candidum extracted liquid (DCEL) for 1 week, saliva and salivary gland biopsies from labial glands of patients were collected and examined by employing immunoreactivity and immunohistochemistry techniques. Results showed that salivary secretion increased by about 65% in patients treated with DCEL as compared with the control group. Higher labeling indices (percentage of acinus area immunoreactive for AQP-5) in the biopsies were found in SS patients who had taken DCEL. This study demonstrated that D. candidum would regulate the expression of AQP-5 in labial glands of SS patients and thereby promoted secretion of saliva to improve dry mouth symptoms.

Polygonatum cyrtonema lectin induces murine fibrosarcoma L929 cell apoptosis via a caspase-dependent pathway as compared to Ophiopogon japonicus lectin - Corrected Proof

2010-07-26

Abstract: Galanthus nivalis agglutinin (GNA)-related lectin family, a superfamily of strictly mannose-binding specific lectins, has been well-known to possess several biological functions including apoptosis-inducing activities. However, the precise mechanisms of GNA-related lectins to induce apoptosis remains to be clarified. In this study, we showed that Polygonatum cyrtonema lectin (PCL) and Ophiopogon japonicus lectin (OJL), the two mannose-binding GNA-related lectins, could induce murine fibrosarcoma L929 cell apoptosis. In addition, we found that there was a close link between their sugar-binding and apoptosis-inducing activities. Interestingly, we further confirmed that the mechanism of lectin-induced apoptosis was a caspase-dependent pathway. Moreover, we found that the two lectins could amplify tumor necrosis factor α (TNFα)-induced apoptosis. Taken together, these findings would open a new perspective for GNA-related lectins as potential anti-tumor agents.

Antioxidant and antiulcerogenic activities of Opuntia ficus indica f. inermis root extract in rats - Corrected Proof

2010-07-20

Abstract: Opuntia ficus indica f. inermis methanolic root extract (ORE) was investigated for phenolic and flavonoids contents, in vitro evaluated for DPPH radical scavenging activity, reducing power and in vivo tested for its gastro-protective ability against 80% ethanol induced ulcer in rats. Phytochemical test of ORE were positive for phenolic and flavonoid contents. DPPH radical scavenging activity and reducing power of ORE showed an EC50 of 118.65±2.51μg/ml and 300μg/ml respectively. In vivo the pre-treatment of rats with ranitidine (50mg/kg) and 200, 400, and 800mg/kg doses of ORE significantly (p<0.05) reduced the 80% ethanol induced-ulcer lesion, with a rate of 82.68%, 49.21%, 83.13%, and 92.59% respectively, and prevented the depletion of antioxidant enzymes, superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), total glutathione (GSH), and inhibited the increase of myeloperoxidase (MPO) and malondialdehyde (MDA) in rat stomach tissues when compared with ethanol group. Also pre-treateatment with ORE marked a dose-dependent attenuation of histopathology changes induced by ethanol. Phenolic and flavonoids wealth, radical scavenging activity, and reducing power, have been implicated for antiulcer property of ORE.

Major tanshinones of Danshen (Salvia miltiorrhiza) exhibit different modes of inhibition on human CYP1A2, CYP2C9, CYP2E1 and CYP3A4 activities in vitro - Corrected Proof

Xin Wang, Ching Mei Cheung, Wayne Y.W. Lee, Penelope M.Y. Or, John H.K. Yeung — 2010-07-20

Abstract: This study investigated the effects of tanshinones on human CYP1A2 (phenacetin O-deethylase), CYP2C9 (tolbutamide 4-hydroxylase), CYP2E1 (chlorzoxazone 6-hydroxylase) and CYP3A4 (testosterone 6β-hydroxylase) activities in vitro using pooled human liver microsomes and specific human CYP isoforms. Tanshinone I, tanshinone IIA, and cryptotanshinone were potent competitive CYP1A2 inhibitors (Ki=1.5–2.5μM); medium competitive inhibitors of CYP2C9 (Ki=22–62μM); medium competitive inhibitors of CYP2E1 (Ki=3.67μM) for tanshinone I and 10.8μM for crytotanshinone; but weak competitive inhibitors of CYP3A4 (Ki=86–220μM). Dihydrotanshinone was a competitive inhibitor of human CYP1A2 (Ki=0.53μM) and CYP2C9 (Ki=1.92μM), a noncompetitive inhibitor of CYP3A4 (Ki=2.11μM) but an uncompetitive CYP2E1 inhibitor. In conclusion, these results showed that tanshinones inhibited the metabolism of various CYP probe substrates in human liver microsomes and specific human CYP isoforms in vitro. Given that CYP1A2, 2C9, 2E1 and 3A4 are responsible for the metabolism and disposition of a large number of drugs currently used, the potential herb–drug interactions of Danshen preparations containing the major tanshinones with drugs which are substrates of these CYPs may be important.

Inhibitory effect of hericenone B from Hericium erinaceus on collagen-induced platelet aggregation - Corrected Proof

2010-07-20

Abstract: Platelet aggregation in the blood vessel causes thrombosis. Therefore, inhibitors of platelet aggregation promise to be preventive or therapeutic agents of various vascular diseases, including myocardial infarction and stroke. In the present study, we found that hericenone B had a strong anti-platelet activity and it might be a novel compound for antithrombotic therapy possessing a novel mechanism. Prior to this study, we examined anti-platelet aggregation activity of ethanol extracts of several species of mushrooms, and found that extract of Hericium erinaceus potently inhibited platelet aggregation induced by collagen. Therefore, we first fractionated the ethanol extract of H. erinaceus to identify the active substances. The anti-platelet activity of each fraction was determined using washed rabbit platelets. As a result, an active component was isolated and identified as hericenone B. Hericenone B selectively inhibited collagen-induced platelet aggregation, but it did not suppress the aggregation induced by U46619 (TXA2 analogue), ADP, thrombin, or adrenaline. Furthermore, hericenone B did not inhibit arachidonic acid- or convulxin (GPVI agonist)-induced platelet aggregation. Therefore, hericenone B was considered to block collagen signaling from integrin α2/β1 to arachidonic acid release. Moreover, we found that collagen-induced aggregation was inhibited by hericenone B in human platelets, similar to in rabbit platelets.

Anti-angiogenic effect of siphonaxanthin from green alga, Codium fragile - Corrected Proof

2010-07-20

Abstract: Since anti-angiogenic therapy has becoming a promising approach in the prevention of cancer and related diseases, the present study was aimed to examine the anti-angiogenic effect of siphonaxanthin from green alga (Codium fragile) in cell culture model systems and ex vivo approaches using human umbilical vein endothelial cells (HUVECs) and rat aortic ring, respectively. Siphonaxanthin significantly suppressed HUVEC proliferation (p<0.05) at the concentration of 2.5μM (50% as compared with control) and above, while the effect on chemotaxis was not significant. Siphonaxanthin exhibited strong inhibitory effect on HUVEC tube formation. It suppressed the formation of tube length by 44% at the concentration of 10μM, while no tube formation was observed at 25μM, suggesting that it could be due to the suppression of angiogenic mediators. The ex vivo angiogenesis assay exhibited reduced microvessel outgrowth in a dose dependent manner and the reduction was significant at more than 2.5μM. Our results imply a new insight on the novel function of siphonaxanthin in preventing angiogenesis related diseases.

Isoflavones isolated from red clover (Trifolium pratense) inhibit smooth muscle contraction of the isolated rat prostate gland - Corrected Proof

A. Brandli, J.S. Simpson, S. Ventura — 2010-07-20

Abstract: This study investigated whether red clover contains any bioactive constituents which may affect contractility of rat prostatic smooth muscle in an attempt to determine whether its medicinal use in the treatment of benign prostatic hyperplasia is supported by pharmacological effects. A commercially available red clover extract was chemically fractionated and various isoflavones (genistein, formononetin and biochanin A) were isolated from these fractions and their effects on contractility were examined on preparations of the isolated rat prostate gland. Contractile effects of the isolated fractions were compared with commercially available isoflavones (genistein, formononetin and biochanin A). Pharmacological tools were used to investigate the mechanism of action modifying smooth muscle contraction. Crude red clover extract (Trinovin®) inhibited electrical field stimulation induced contractions of the rat prostate across a range of frequencies with an IC50 of approximately 68μg/ml. Contractions of the rat prostate elicited by exogenous administration of acetylcholine, noradrenaline or adenosine 5′-triphosphate (ATP) were also inhibited. Chromatographic separation, and final purification by high performance liquid chromatography (HPLC) permitted the isolation of the isoflavones: daidzein, calycosin, formononetin, prunetin, pratensin, biochanin A and genistein. Genistein, formononetin and biochanin A (100μM) from either commercial sources or isolated from red clover extract inhibited electrical field stimulation induced contractions of the isolated rat prostate. It is concluded that isoflavones contained in red clover are able to inhibit prostatic smooth muscle contractions in addition to their antiproliferative effects. However, the high concentrations required to observe these smooth muscle relaxant effects mean that a therapeutic benefit from this mechanism is unlikely at doses used clinically.

A pharmacodynamic–pharmacokinetic (PD–PK) study on the effects of Danshen (Salvia miltiorrhiza) on midazolam, a model CYP3A probe substrate, in the rat - Corrected Proof

Xin Wang, Wayne Y.W. Lee, Xuelin Zhou, Penelope M.Y. Or, John H.K. Yeung — 2010-07-20

Abstract: This study investigated the effect of Danshen on the pharmacodynamic–pharmacokinetic (PD–PK) effects of midazolam, a model CYP3A probe substrate. The effects of acute and 3-day Danshen treatment on the pharmacokinetics of a low dose midazolam (10mg/kg, i.p.) were determined in vivo in the rat. Danshen (200mg/kg, i.p.) treatment decreased midazolam clearance by 16%, with increases in the AUC by 22% and the half-life by 14%. 3-Day Danshen treatment (200mg/kg/day, i.p.) for 3 days decreased the clearance, with increases in the T1/2 and AUC. The effects of acute and 3-day Danshen on midazolam-induced hypnosis, serum 1′-hydroxy-midazolam to midazolam ratio and hepatic CYP3A protein expression were determined in the rat. Danshen treatments (100–200mg/kg, i.p. and 200–500mg/kg, p.o.) increased the sleeping time (p<0.001) produced by a hypnotic dose of midazolam (50mg/kg, i.p.) without affecting the sleep latency. Serum 1′-hydroxy-midazolam to midazolam ratio after the hypnotic dose of midazolam was decreased after intraperitoneal Danshen treatment (200mg/kg) but not after oral treatment at up to 500mg/kg. All the treatment groups with Danshen, after intraperitoneal and oral administration, decreased hepatic CYP3A protein expression (p<0.05) by about 25%. The results confirmed that Danshen had no enzyme inducing effects on rat CYP3A.

Synergistic effect of the interaction between naproxen and citral on inflammation in rats - Corrected Proof

2010-07-20

Abstract: The combination of non-steroidal anti-inflammatory drugs with herbs having analgesic effects can increase their antinociceptive activity and limit their side effects. The aim of the present study was to examine the effects on inflammation and gastric injury in rats resulting from the interaction between naproxen and citral. Naproxen, citral, or fixed-dose naproxen–citral combinations were administered orally and their anti-inflammation (carrageenan-induced paw edema) and gastric damage were assessed in rats. The pharmacological interaction type was evaluated by the isobolographic analysis. Naproxen, citral, or combinations of naproxen and citral produced anti-inflammatory effects. The sole administration of naproxen produced significant gastric damage, but this effect was not obtained with either citral or combinations. ED30 values were estimated for the individual drugs, and isobolograms were constructed. The derived theoretical ED30 for the anti-inflammatory effect was 504.4mg/kg; this was significantly higher than the observed experimental value (190.6mg/kg). These results indicate that a synergistic interaction underlies the anti-inflammatory effect. The data suggests that the naproxen–citral combination can interact and to produce minor gastric damage and may have therapeutic advantages for the clinical treatment of inflammation.

Curcumin improves bone microarchitecture and enhances mineral density in APP/PS1 transgenic mice - Corrected Proof

2010-07-20

Abstract: Alzheimer's disease and osteoporosis are often observed to co-occur in clinical practice. The present study aimed to evaluate the bone microarchitecture and bone mineral density (BMD) of the proximal tibia in APP/PS1 transgenic mice by micro-computed tomography (micro-CT), and to search for evidence that curcumin can be used to reduce bone mineral losses and treat osteoporosis after senile dementia in these transgenic mice. Three-month-old female mice were divided into the following groups (n=9 per group): wild-type mice (WT group); APP/PS1 transgenic mice (APP group); and APP/PS1 transgenic mice with curcumin treatment (APP+Cur group). Between 9 and 12 months of age, the APP+Cur group were administered curcumin orally (600ppm). CT scans of the proximal tibia were taken at 6, 9 and 12 months. At 6 months, there were little differences in the structural parameters. At 9 months, the APP groups displayed loss of bone volume ratio (BV/TV), trabecular thickness (Tb.Th), trabecular number (Tb.N) and connectivity density (Conn.D) and increases in trabecular separation (Tb.Sp) and geometric degree of anisotropy (DA) (P<0.05 or P<0.01), with significant changes in the BMD parameters. At 12 months, curcumin treatment led to constant increases in the trabecular bone mass of the metaphysis and clearly improved the BMD. By the same time, we measured the TNF-α and IL-6 in the serum among the different groups at 6, 9 and 12 months by enzyme-linked immunoassay(ELISA). These results suggest that APP/PS1 transgenic mice are susceptible to osteoporosis, and that curcumin can prevent further deterioration of the bone structure and produce beneficial changes in bone turnover. The change of inflammation cytokine, including TNF-α and IL-6, may play an important role in the mechanisms of action of curcumin, but the detail mechanism remains unknown.

A comparative chemical and pharmacological study of standardized extracts and vanillic acid from wild and cultivated Amburana cearensis A.C. Smith - Corrected Proof

2010-07-20

Abstract: The objectives of this work were to carry out a comparative chemical study and to evaluate the antinociceptive and anti-inflammatory activities of ethanol extracts (EtOHE) and vanilic acid (VA) from cultivated and wild Amburana cearensis A.C. Smith (Fabaceae), an endangered species used in Northeast Brazil for the treatment of asthma. The HPLC analysis of EtOHE, showed that coumarin (CM) and VA were the major constituents from the cultivated plant, while in the extract from the wild plant the major constituents were amburoside A (AMB) and CM. Pharmacological tests were performed with male Swiss mice or male Wistar rats acutely administered with 100–400mg/kg, p.o. of EtOHEs or 12.5–50mg/kg, p.o. of VA. EtOHEs from A. cearensis with 4, 7 or 9 months of cultivation significantly inhibited, from 32 to 64%, both phases of the formalin test in mice. Similar results were observed with the EtOHE from the wild species. VA significantly reduced both phases of the formalin test. This effect was partially reversed by naloxone. EtOHE from cultivated or wild A. cearensis inhibited the carrageenan (Cg)-induced mice paw edema. Furthermore, VA inhibited the paw edema and the leukocyte migration in rat peritoneal cavity induced by Cg. On the other hand, it did not inhibit the edema and the increase of vascular permeability induced by dextran in the rat paw. All together, these results indicate that the EtOHE from cultivated A. cearensis exhibit similar chemical and pharmacological profiles, as related to the wild plant. VA is, at least partially, responsible for these pharmacological effects. Its antinociceptive effect occurs by a mechanism partly dependent upon the opioid system, while the anti-inflammatory action was manifested in inflammatory processes dependent on polymorphonuclear cells and are probably related to the VA inhibition of cytokines as observed by others.

Analgesic effects of the ethanolic extract from Magnolia ovata (Magnoliaceae) trunk bark and of N-acetylxylopine, a semi-synthetic analogue of xylopine - Corrected Proof

2010-07-20

Abstract: This study investigated the antinociceptive effects of the ethanolic extract (EEMO) obtained from Magnolia ovata (A.St.-Hil.) Spreng and N-acetylxylopine (AXyl), a stable derivative of xylopine in different models of nociception. The EEMO and AXyl inhibited the nociception induced by acetic acid in mice, in a dose-dependent manner with a maximal inhibition of 91±9% and 50±11%, respectively. Oral administration of EEMO or AXyl also significantly inhibited the inflammatory phase of formalin-induced nociception with maximal reduction of 87±3.9% and 71±10%, respectively. Confirming the effectiveness of the extract and the isolated compound in inflammatory responses, EEMO or AXyl inhibited carrageenan-induced mechanical allodynia with percentage of inhibition of 40±6% for EEMO and 82±8% for AXyl. Intraplantar injection of AXyl in the ipsilateral paw, but not in the contralateral paw, also reduced carrageenan-induced mechanical allodynia in mice. The response of the animals for maximal doses tested of EEMO and AXyl in the hot-plate or rota-rod models were not altered. These results show that the extract from M. ovata and the stable derivative AXyl possess analgesic properties towards inflammatory pain acting on peripheral sites.

Tanshinone II-A attenuates cardiac fibrosis and modulates collagen metabolism in rats with renovascular hypertension - Corrected Proof

2010-07-20

Abstract: The adaptive changes that develop in the pressure-overloaded left ventricular myocardium include cardiac hypertrophy and interstitial fibrosis. The objectives of the present study were to evaluate the effects of Tanshinone II-A, a bioactive diterpene quinone isolated from Danshen, on cardiac fibrosis and collagen metabolism in rats with renovascular hypertension. Male Sprague-Dawley rats were subjected to two-kidney two-clip (2K2C) or sham operation (sham) and treated with Valsartan (Val, 26.7mg/kg/d), Tanshinone II-A (Tsn, 70, 35mg/kg/d) or vehicle. Six weeks later, systolic blood pressure (BP), LV weight, collagen abundance, cardiac function parameters, hydroxyproline content and mRNA levels of matrix metalloproteinase (MMP)-2, MMP-9, tissue inhibitor of metalloproteinase (TIMP)-1 and TIMP-2 were evaluated. Both high-dose (Tsn-H, 70mg/kg/d) and low-dose (Tsn-L, 35mg/kg/d) of Tsn failed to attenuate 2K2C-induced BP elevation but significantly attenuated the attendant interstitial fibrosis. Val suppressed elevations of BP and left ventricular systolic pressure (LVSP) in 2K2C rats. Val and Tsn-H exerted comparable suppressive effects on the gene expression of MMP-9 and TIMP-1, while Val decreased the MMP-2 mRNA level without affecting the transcript levels of TIMP-2. Both Val and Tsn-H attenuated cardiac dysfunction, while Tsn-L showed slight improvement. These data demonstrate for the first time, that Tsn prevented cardiac fibrosis and improved cardiac function in a rat model of renovascular hypertensive independent of hypotensive effect. Tsn conferred its beneficial effects on the collagen metabolism probably through its regulation of transcript levels of the MMPs/TIMPs balance.

Moringa oleifera induced potentiation of serotonin release by 5-HT3 receptors in experimental ulcer model - Corrected Proof

Siddhartha Debnath, Debasis Biswas, Koushik Ray, Debjani Guha — 2010-07-20

Abstract: Ethnopharmacological relevance: Moringa oleifera (Moringaceae), a perennial plant is widely cultivated throughout the world. Extensive pharmacological studies revealed its promising role in modulation of various disorders like antispasmodic, diuretic, abortifacient, antimicrobial antibacterial, antitubercular, antiviral, antifertility, depressant, anti-inflammatory and anticancer property which promoted us to conduct the study to elucidate its role on experimental gastric ulceration.Aim of the study: The aim of the present study was to assess the efficacy of its aqueous leaf extract on protection of gastric ulceration and characterize the possible modulatory mechanism underlying the phenomenon.Materials and methods: Adult Holtzman strain albino rats (weight 150–200g) of either sex were used for the study. Ulceration was induced using aspirin (500mg/kg body weight) and using Moringa oleifera (MO), a herbal formulation, the modulatory mechanism has been studied and compared with a commonly used antagonist of 5-HT3 receptors, ondansetron by assessing parameters like mean ulcer index, 5-HT content, EC cell count and mucosal thickness.Results: The results of our study suggest that MO protects ulcer formation by modulating 5-HT secretion through EC cell via 5-HT3 receptors in gastrointestinal tract.Interpretation and conclusion: MO showed maximum protective activity at a dose of 300mg/kg body weight against above-mentioned experimental rat ulcer model by modulating 5-HT secretion through EC cell via 5-HT3 receptors in gastrointestinal tract which has given a glimpse of a therapeutic approach for gastric ulcer management, which may be beneficially used in contrast to the classical antacid, antihistamine or surgical treatment. Further investigations and proper screening regarding various phytochemicals, alkaloids present within MO leaf will help to formulate effective herbal preparation that will be used to combat gastrointestinal disorders in future.

Flavonoids, apigenin and icariin exert potent melanogenic activities in murine B16 melanoma cells - Corrected Proof

Yan Ye, Gui-Xin Chou, Hui Wang, Jian-Hong Chu, Zhi-Ling Yu — 2010-07-20

Abstract: We aimed to screen for melanogenic agents among 35 botanical compounds. The compounds were first assessed with regard to their effects on tyrosinase activity in B16 cells. At 100μM, 13 compounds showed tyrosinase activity-enhancing effects, ranging from 2.6 to 372.8% activation. Five of them showed more than 50% enhancement and were further tested for their EC50 values. Compared with 8-Methoxypsoralen, an effective tyrosinase activator with an EC50 of 7.26μM, 3 compounds exhibited smaller EC50 values (apigenin, 0.45μM; hyperosid, 0.92μM; and icariin, 1.01μM for enhancing tyrosinase activity). The 3 compounds significantly increased cellular melanin contents without affecting cell proliferation. Compared with 8-Methoxypsoralen (EC50, 35.94μM for stimulating pigmentation), apigenin (EC50, 17.46μM) and icariin (EC50, 32.77μM) showed better melanogenic activity, while hyperosid (EC50, 70.4μM) was less potent. Western blot analysis demonstrated that the 3 compounds could differentially increase the expression levels of tyrosinase, and tyrosinase-related proteins 1 and 2. Together these data suggest that apigenin and icariin exert potent melanogenic activities through, at least in part, upregulating the protein expression levels of melanogenic enzymes in B16 cells. Thus, further investigations are merited to ascertain their potential application in treating hypopigmentation disorders.

Taiwanin A inhibits MCF-7 cancer cell activity through induction of oxidative stress, upregulation of DNA damage checkpoint kinases, and activation of p53 and FasL/Fas signaling pathways - Corrected Proof

2010-07-20

Abstract: This study investigates the anti-MCF-7 breast cancer cell effects and the underlying pharmacological activity and mechanism of taiwanin A, a major lignan isolated from Taiwania cryptomerioides. Our results show that taiwanin A time-dependently induced reactive oxygen species level and DNA damage in MCF-7 cells, which were likely activated kinases ataxia telangiectasia mutated (ATM) and checkpoint kinase (Chk). Taiwanin A could also up-regulate p53, phosphorylated p53, p21Cip1, and p27Kip1 and down-regulate the G2/M checkpoint cyclin-dependent kinase1 (Cdk1)-cyclin A/B, leading to induction of G2/M cell-cycle arrest in MCF-7 cells. Blockade of p53 gene expression by siRNA further demonstrated that the cell-cycle arrest induced by taiwanin A was p53-dependent. The FasL/Fas-mediated apoptotic signaling cascade was involved in taiwanin A-induced apoptosis via activation of caspases-10 and -7 (but not caspase-8), and proteolytic cleavage of poly(ADP-ribose) polymerase (PARP). In contrast, mitochondria-initiated apoptotic pathway was not involved. This is the first report to delineate novel mechanism of the action of taiwanin A against MCF-7 cells, suggesting this lignan may have value for development as an anti-breast cancer agent.

Cardioprotective activity of Cladosiphon okamuranus fucoidan against isoproterenol induced myocardial infarction in rats - Corrected Proof

Paul Thomes, Murugan Rajendran, Balu Pasanban, Ramasamy Rengasamy — 2010-07-20

Abstract: Fucoidans, sulfated polysaccharides of brown algae, have attracted steady attention in the last few years as readily accessible biopolymers possessing a wide spectrum of biological activities. In this study, cardioprotective activity of fucoidan extracted from Cladosiphon okamuranus was evaluated in isoproterenol induced myocardial infarction in rats. Male Wistar albino rats (180±25g) were divided in to four groups of six animals each as follows: Group (1) control, Group (2) isoproterenol alone, Group (3) fucoidan alone and Group (4) fucoidan+isoproterenol. To evaluate the efficacy of fucoidan treatment against isoproterenol induced myocardial damage, biochemical parameters and histopathological studies were carried out. Isoproterenol administration produced severe myocardial damage and high lipid peroxidation level. On the contrary, fucoidan treatment reduced myocardial damage, which has been reflected by improvement in parameters such as creatinine phosphokinase (CPK), lactate dehydrogenase (LDH), alanine transaminase (ALT) and aspartate transaminase (AST). In addition, fucoidan improved the antioxidant defence system in treated animals and considerably reduced the oxidative stress exerted by isoproterenol. The reduction in oxidative stress in Group (4) was evident from the lipid peroxidation and antioxidant activities. Furthermore, the increase in the levels of total cholesterol, triglycerides and low density lipoprotein (LDL) and decrease in the levels of high density lipoprotein (HDL) was significantly reversed in Group (4), when compared with Group (2). The histopathological studies also showed that fucoidan treatment significantly minimized the damage induced by isoproterenol. Thus, fucoidan provide cardioprotection against isoproterenol induced myocardial infarction in rats.

Antiedematogenic activity and phytochemical composition of preparations from Echinodorus grandiflorus leaves - Corrected Proof

2010-07-16

Abstract: The leaves of Echinodorus grandiflorus (Alismataceae) are traditionally used in Brazil to treat inflammatory conditions. The aim of the present study was to evaluate the antidematogenic activity of crude aqueous, dichloromethane and hydroethanolic extracts from E. grandiflorus leaves using the carrageenan-induced paw edema model in mice, along with of fractions enriched in diterpenes, flavonoids and hydroxycinnamoyltartaric acids (HCTA). Significant inhibitions of paw edema were elicited by the 50% and 70% EtOH extracts (1000mg/kg, p.o.), as well as by the fractions enriched in diterpenes (70–420mg/kg, p.o.) and flavonoids (7.2–36mg/kg, p.o.). Isovitexin, isoorientin, trans-aconitic and chicoric acids were identified in all extracts by HPLC analysis. Trans-aconitic acid itself exhibited significant antiedematogenic effect (270mg/kg, p.o.). The biological activity correlated positively with the contents of flavonoids and diterpenes, but negatively with HCTA concentrations, demonstrating the participation of the two classes of compounds in the antiedematogenic activity of E. grandiflorus.

Pre-treatment with α-hederin increases β-adrenoceptor mediated relaxation of airway smooth muscle - Corrected Proof

Anne Wolf, Reinoud Gosens, Herman Meurs, Hanns Häberlein — 2010-07-16

Abstract: Preparations of ivy leaves dry extract with secretolytic and bronchiolytic efficacy are widely used for the treatment of acute and chronic obstructive airway diseases. The mechanism by which ivy preparations improve lung functions is not fully understood. Here, we tested the influence of the three main saponins of ivy, α-hederin, hederacoside C and hederagenin, on the contraction and relaxation behaviour of isolated bovine tracheal smooth muscle strips by isometric tension measurements. None of the tested compounds altered histamine or methacholine-induced contraction of the smooth muscle strips. In contrast, the isoprenaline-induced relaxation of 100μM methacholine precontracted muscle strips was significantly enhanced when pre-treated with 1μM of α-hederin for 18h. The pre-treatment with hederacoside C or hederagenin had no effect on isoprenaline-induced relaxation. For the first time the bronchiolytic effect of α-hederin was demonstrated by isometric tension measurements using bovine tracheal smooth muscle strips. α-Hederin increases isoprenaline-induced relaxation indirectly, probably by inhibiting heterologous desensitization induced by high concentrations of muscarinic ligands like methacholine.

Ursolic acid and oleanolic acid, members of pentacyclic triterpenoid acids, suppress TNF-α-induced E-selectin expression by cultured umbilical vein endothelial cells - Corrected Proof

K. Takada, T. Nakane, K. Masuda, H. Ishii — 2010-06-25

Abstract: E-selectin is an early response adhesion molecule expressed on the surface of endothelial cells during inflammatory responses. We examined the effects of two pentacyclic triterpenoid acids, ursolic acid (UA) and oleanolic acid (OA), on the expression of E-selectin by cultured human umbilical vein endothelial cells (HUVECs). Treatment of the cells with UA or OA alone did not influence expression of E-selectin. Expression of E-selectin mRNA and surface antigen by HUVECs was induced by treatment with tumor necrosis factor-α (TNF-α) in a dose- and time-dependent manner. TNF-α-induced up-regulation of E-selectin was abrogated by pre-treatment of the cells with UA or OA which decreased expression of E-selectin mRNA. The repression of E-selectin mRNA expression caused by the pentacyclic triterpenoid acids paralleled the inhibition of NF-κB activation and nuclear translocation, as evaluated by electrophoretic mobility shift assays, although the degree of repression by UA was approximately two times more effective than that of OA. The results suggest that UA and OA suppress the inflammatory cytokine-induced expression of E-selectin in endothelial cells by decreasing E-selectin transcription via inhibition of NF-κB activation. Thus, UA and OA function as anti-inflammatory agents. The differences in the inhibitory efficacy between UA and OA may be due to conformational differences in ring-E of the two pentacyclic triterpenoid acids.

Improvement of signs and symptoms of chronic venous insufficiency and microangiopathy with Pycnogenol®: A prospective, controlled study - Corrected Proof

2010-06-25

Abstract: The aim of this study was to evaluate the clinical efficacy of standardized French maritime pine bark extract Pycnogenol® in patients with severe chronic venous insufficiency (CVI). 98 subjects with symptomatic CVI and edema were randomly assigned to one group treated with 150mg Pycnogenol a day only, another group with stockings only and a third group with both Pycnogenol and elastic stockings. The average ambulatory venous pressure (AVP) at inclusion was 58±7mmHg (range 48–60mmHg) with a refilling time (RT)<12s (average 7±2s). The duration of the disease was on average 6.0±3.1 years. There were no differences in AVP or RT among the 3 groups at inclusion and microcirculatory and clinical evaluations were comparable.After 8 weeks treatment there was a significant decrease of rate of ankle swelling, resting flux, transcutaneous pO2 and clinical symptom scores in all groups with significantly better results for the combination treatment. Pycnogenol alone was more effective than compression alone for all parameters (p<0.05). No side-effects were observed; compliance and tolerability were very good.This study corroborates a significant clinical role for Pycnogenol in the management, treatment and control of CVI also in combination with compression.

Effect of silymarin on kidneys of rats suffering from alloxan-induced diabetes mellitus - Corrected Proof

C. Soto, J. Pérez, V. García, E. Uría, M. Vadillo, L. Raya — 2010-06-25

Abstract: Oxidative stress contributes to the pathogenesis of diabetes mellitus and its sequelae nephropathy. The kidneys are especially prone to damage by free radicals. We therefore tested the effect of the flavonoid mixture silymarin, a free radical scavenger, on the activity and gene expression of superoxide dismutase, glutathione peroxidase and catalase, as well as on renal tissue morphology in rats with alloxan-induced diabetes mellitus. Alloxan-intoxicated rats were treated with silymarin 20 days after alloxan administration for 9 weeks. Alloxan-induced tissue damage and decreased the activity of the three enzymes, SOD (U/mgprot.): 14.4±1.75 vs 112±6.45 control, p<0.05, n=6; GSHPx (μMNADPH/min/mgprot.): 0.02±0.002 vs 0.121±0.01 control, p<0.05, n=6; CAT (k/seg/mgprot.): 0.022±0.003 vs 0.044±0.002 control, p<0.05, n=6. Silymarin treatment prevented tissue damage and restored the activity (SOD: 110.7±12.9U/mg prot.; GSHPx: 0.329±0.031μMNADPH/min/mgprot.; CAT: 0.054±0.002k/seg/mgprot., n=6) and gene expression of the three antioxidant enzymes after 20 days of alloxan administration (SOD: 12.00±0.57 control, 9.00±0.1 diabetic p<0.05, 11.00±0.20 silymarin treated; GSHPx: 6.01±0.78 control, 9.03±0.3 diabetic p<0.05, 7.02±0.07 silymarin treated; CAT: 9.03±1.07 control, 12.02±0.60 diabetic p<0.05, 8.06±0.31 silymarin treated, n=6). It is suggested in this study that recuperative effect of silymarin on the renal tissue damage induced by alloxan may be related to an increase in the activity and recovery of gene expression of antioxidant enzymes which in addition to the glutathione system constitute some of the most important defense mechanisms against free radicals damage. As these results show, silymarin may be considered potentially in the treatment of diabetic nephropathy.

Anticancer activity of the Uncaria tomentosa (Willd.) DC. preparations with different oxindole alkaloid composition - Corrected Proof

2010-06-25

Abstract: The activity of Uncaria tomentosa preparations on cancer cells was studied using in vitro and in vivo models. IC50 values were calculated for preparations with different quantitative and qualitative oxindole alkaloid composition: B/W37 – bark extracted in water at 37°C, B/Wb – bark extracted in boiling water, B/50E37 – bark extracted in 50% ethanol at 37°C, B/Eb – bark extracted in boiling 96% ethanol, B/96E37 – bark extracted in 96% ethanol at 37°C and B/SRT – bark extracted in water and dichloromethane. Generally, the results obtained showed a high correlation between the total oxindole alkaloid content (from 0.43% to 50.40% d.m.) and the antiproliferative activity of the preparations (IC50 from >1000μg/ml to 23.57μg/ml). B/96E37 and B/SRT were the most cytotoxic preparations, whereas the lowest toxicity was observed for B/W37. B/96E37 were shown to be active against Lewis lung carcinoma (LL/2) [IC50=25.06μg/ml], cervical carcinoma (KB) [IC50=35.69μg/ml] and colon adenocarcinoma (SW707) [IC50=49.06μg/ml]. B/SRT was especially effective in inhibiting proliferation of cervical carcinoma (KB) [IC50=23.57μg/ml], breast carcinoma (MCF-7) [IC50=29.86μg/ml] and lung carcinoma (A-549) [IC50=40.03μg/ml]. Further animal studies on mice bearing Lewis lung carcinoma showed significant inhibition of tumor growth by B/W37 administered for 21 days at daily doses of 5 and 0.5mg (p=0.0009). There were no significant changes in the cell cycles of tumor cells with the exception of cell decrease at the G2/M phase after the administration of B/96E37 at a daily dose of 0.5mg and the G1/G0 cells cycle arrest demonstrated after the B/SRT therapy at a daily-dose of 0.05mg. All tested preparations were non-toxic and well tolerated.

S-allylcysteine reduces the MPTP-induced striatal cell damage via inhibition of pro-inflammatory cytokine tumor necrosis factor-α and inducible nitric oxide synthase expressions in mice - Corrected Proof

2010-06-24

Abstract: We have recently demonstrated that S-allylcysteine (SAC) induces protection on neurochemical, biochemical and behavioral markers of striatal damage in different neurotoxic animal models – including a murine model induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropiridinium (MPTP) injection to mice – indicating that pro-oxidant reactions underlie neurotoxicity in these models (). In this work we investigated whether SAC can protect the striatum of mice from the morphological alterations in the MPTP toxic model, and if this response is correlated with a reduction in pro-inflammatory cytokine tumor necrosis factor-α (TNF-α) and inducible nitric oxide synthase (iNOS) expressions, and further reduction in astrocyte activation (glial fibrillary acidic protein (GFAP) expression). The striatal tissue from MPTP injected animals (30mg/kg, i.p., ×5 days) showed a significant degree of cell damage and enhanced immunoreactivities to GFAP, TNF-α and iNOS, as well as an enhanced number of apoptotic nuclei. Treatment of mice with SAC (120mg/kg, i.p., ×5 days) in parallel to MPTP significantly reduced or prevented all these markers. Our results suggest that MPTP-induced morphological alterations recruit a pro-inflammatory component triggered by cytokine TNF-α release and nitric oxide formation, which is sensitive to the antioxidant properties of SAC. This antioxidant is an effective experimental tool to reduce the brain lesions associated with oxidative damage and inflammatory responses.

Chemical composition of the SFE-CO2 extracts from Cajanus cajan (L.) Huth and their antimicrobial activity in vitro and in vivo - Corrected Proof

Yuan-gang Zu, Xiao-lei Liu, Yu-jie Fu, Nan Wu, Yu Kong, Michael Wink — 2010-06-24

Abstract: The in vitro and in vivo antimicrobial activities of SFE-CO2 (supercritical fluid extraction) extracts and ethanol extracts from Cajanus cajan (L.) Huth were investigated. The flavonoid compounds orientin, vitexin, isovitexin, pinostrobin and the stilbene cajaninstilbene acid were detected in SFE-CO2 extracts by HPLC-DAD. In vitro antimicrobial activities of the extracts were evaluated against eight microbial strains (the bacteria Staphylococcus epidermidis, Staphylococcus aureus, Bacillus subtilis, Proteus vulgaris, Pseudomonas aeruginosa, Escherichia coli; and the fungi Aspergillus niger and Candida albicans). A marked inhibitory effect of the SFE extracts was observed against Staphylococcus epidermidis, Staphylococcus aureus and Bacillus subtilis. The IC50 of SFE-CO2 extracts ranged from 0.0557mg/ml to 0.0689mg/ml consisting of cancer (MCF-7 (0.0557mg/ml)) as well as non-cancer (BHK-21 (0.0641mg/ml), RAW264.7 (0.0689mg/ml) and Vero (0.0625mg/ml)) cells.Flow cytometry (FCM) was used to analyze death rate of the most sensitive strain (Staphylococcus aureus) caused by the SFE extracts. Additionally, the whole cell proteins of Staphylococcus aureus were analyzed by SDS-PAGE to detect if there were changes in protein patterns. In vivo antimicrobial activity was studies in mice that had been inoculated with Staphylococcus aureus. The potential mechanism of antimicrobial activity in vivo was studied by histopathology.

Effect of combined administration of ginger (Zingiber officinale Roscoe) and atorvastatin on the liver of rats - Corrected Proof

Gehan H. Heeba, Manal I. Abd-Elghany — 2010-06-24

Abstract: Ginger is known to possess hypolipidemic, antioxidant and hepatoprotective properties. Combination therapy often takes advantage of complementary effects of different agents. This study investigated the combined effect of ginger extract (GE) and atorvastatin on lipid profile and on atorvastatin-induced hepatic injury. Rats were randomized into: control; GE (400mg/kg); atorvastatin (20mg/kg) alone or with GE or vitamin E, and atorvastatin (80mg/kg) alone or with GE or vitamin E. Administration of 80mg/kg atorvastatin for 4 weeks had major hepatotoxic effect whereas the lower dose (20mg/kg) seems to cause mild liver injury. Besides lowering serum total cholesterol and hepatic superoxide dismutase (SOD) and catalase (CAT), atorvastatin significantly increased serum aminotransferases, hepatic malondialdehyde (MDA) and nitric oxide (NO). Concurrent administration of GE and atorvastatin had the opposite effect. Histopathological study revealed that GE reduced liver lesions induced by atorvastatin. The results indicate that the ability of ginger to lower serum cholesterol and to decrease aminotransferases, MDA and NO is clinically important, because its chronic administration will neither lead to side-effects nor to hepatic changes as occurs with high atorvastatin doses. Therefore, combination regimens containing GE and low dose of statins could be advantageous in treating hypercholesterolemic patients which are susceptible to liver function abnormalities.

Antidiabetic effect of S-allylcysteine: Effect on plasma and tissue glycoproteins in experimental diabetes - Corrected Proof

G. Saravanan, P. Ponmurugan, G.P. Senthil Kumar, T. Rajarajan — 2010-06-24

Abstract: The present study was conducted to investigate the effect of S-allylcysteine (SAC) on dearrangement in glycoprotein levels in the streptozotocin induced diabetic model. SAC (150mg/kg b.w./day) was administered orally for 45 days to normal and diabetic rats. STZ-induced diabetic rats showed significant increase in blood glucose and glycoprotein components such as hexose, hexosamine, fucose and sialic acid in plasma, liver and kidneys of diabetic rats. Oral administration of SAC to diabetic rats for a period of 45 days normalized all the above-mentioned biochemical parameters. The antihyperglycemic effect of SAC was compared with glyclazide, a well-known antihyperglycemic drug. The present study indicates that SAC possesses a significantly beneficial effect on the glycoprotein moiety in addition to its antidiabetic effect.

Isolation of adenosine, iso-sinensetin and dimethylguanosine with antioxidant and HIV-1 protease inhibiting activities from fruiting bodies of Cordyceps militaris - Corrected Proof

2010-06-24

Abstract: According to previous studies, a close relationship between oxidative stress and AIDS suggests that antioxidants might play an important role in the treatment of AIDS. Cordyceps militaris was selected from nine edible mushrooms by assay of inhibition of erythrocyte hemolysis. Macroporous adsorption resin and HPLC were used to purify three micromolecular compounds named L3a, L3b and L3c. L3a was identified to be adenosine with the molecular formula C10H13N5O4; L3b was 6,7,2′,4′,5′-pentamethoxyflavone with the molecular formula C20H20O7, and L3c was dimethylguanosine with the molecular formula C12H17N5O5. The compound 6,7,2′,4′,5′-pentamethoxyflavone was first isolated from C. militaris. The assay of inhibition of HIV-1 protease (HIV-1 PR) was based on the fact that the expression of this enzyme can inhibit the growth of E. coli. This is a new screening system for HIV-1 PR inhibitors. Both L3a and L3b showed high inhibition to HIV-1 PR. These compounds could be new anti-HIV-1 PR drugs.

Anthelmintic activity of steroidal saponins from Paris polyphylla - Corrected Proof

G.-X. Wang, J. Han, L.-W. Zhao, D.-X. Jiang, Y.-T. Liu, X.-L. Liu — 2010-06-24

Abstract: The present study was undertaken to investigate the anthelmintic activity of crude extracts and pure compounds from the rhizomes of Paris polyphylla. The methanol extract showed a promising anthelmintic activity against Dactylogyrus intermedius (EC50 value=18.06mgl−1). Based on these finding, the methanol extract was fractionated on silica gel column chromatography in a bioassay-guided fractionation affording two known steroidal saponins showing potent activity, dioscin (1) and polyphyllin D (2). Both dioscin and polyphyllin D exhibited significant activity against D. intermedius with EC50 values of 0.44 and 0.70mgl−1, respectively, which were more effective than the positive control, mebendazole (EC50 value=1.25mgl−1). The acute toxicities (LC50) of dioscin and polyphyllin D for goldfish were 1.37 and 1.08mgl−1, respectively. These results indicated that P. polyphylla extract and the isolated compounds are potential natural agents for the control of Dactylogyrus infestation. This is the first report on in vivo anthelmintic investigation for P. polyphylla.

Anticonvulsant activity of embelin isolated from Embelia ribes - Corrected Proof

S. Mahendran, B.S. Thippeswamy, V.P. Veerapur, S. Badami — 2010-06-02

Abstract: Anticonvulsant activity of embelin (2.5, 5 and 10mg/kg, i.p.) was studied. It showed a significant inhibition of the seizures induced by electroshock and pentylenetetrazole in a dose dependent manner and the activity was comparable to phenytoin and diazepam. Significant decrease in locomotion revealing its CNS depressant activity was observed. The findings suggest that embelin possess anticonvulsant activity against both grand mal and petit mal epilepsy.

Effect of crocetin from Gardenia Jasminoides Ellis on sleep: A pilot study - Corrected Proof

H. Kuratsune, N. Umigai, R. Takeno, Y. Kajimoto, T. Nakano — 2010-06-01

Abstract: Crocetin is a pharmacologically active carotenoid compound of Gardenia jasminoides Ellis used as a traditional herbal medicine and natural colorant. The present pilot study investigated the effect of crocetin on sleep. The clinical trial comprised a double-blind, placebo-controlled, crossover trial of 21 healthy adult men with a mild sleep complaint. It included two intervention periods of 2 weeks each, separated by a 2-week washout period. We measured objective sleep quality using an actigraph, and assessed the subjective symptoms using St Mary's Hospital Sleep Questionnaire. Actigraph data showed that after administration of crocetin, the number of wakening episodes was reduced compared to that of the placebo (p=0.025). Subjective data from St Mary's Hospital Sleep Questionnaire showed that crocetin tended to improve the quality of sleep compared to sleep before its intake. Additionally, no side effects from crocetin intake were observed. The results suggest that crocetin may contribute to improving the quality of sleep.

12-O-Tetradecanoyl phorbol-13-acetate (TPA)-induced growth arrest is increased by silibinin by the down-regulation of cyclin B1 and cdc2 and the up-regulation of p21 expression in MDA-MB231 human breast cancer cells - Corrected Proof

2010-05-31

Abstract: TPA is a potent regulator of cell growth, including cell proliferation and differentiation. In this study, we determined the effect of silibinin on TPA-induced growth arrest in breast cancer cells. Silibinin increased growth arrest of the G2/M phase in a dose-dependent fashion. Silibinin decreased the basal level of cyclin B1 and cdc2 expression, which is involved in S phase and G2/M transition. In addition, TPA-induced G2/M phase arrest was increased by silibinin. Under the same conditions, TPA-induced down-regulation of cyclin B1 and cdc2 was decreased by silibinin. In contrast, TPA-induced p21 expression was further increased by silibinin. To determine the regulatory mechanism of TPA-induced growth arrest, we pretreated cells with various inhibitors, such as UO126, SB203580, and LY294002. Interestingly, TPA-induced growth arrest was significantly increased by LY294002, but not by UO126 and SB203580. In addition, TPA-induced down-regulation of cyclin B1 was inhibited by LY294002; however, the basal level of p21 was increased by TPA and TPA-induced p21 expression was further increased by LY294002. Finally, adenoviral constitutively active-Akt (Ad-CA-Akt) overexpression regulated the up-regulation of cyclin B1 and the down-regulation of p21. Therefore, we have demonstrated that silibinin has an additive effect on TPA-induced growth arrest through the PI-3-kinase/Akt-dependent pathway.

Anti-osteoporotic constituents from Indian medicinal plants - Corrected Proof

2010-05-31

Abstract: The objective of this study was to determine the in vitro osteogenic activities of selected medicinal plants used traditionally in India. The compounds isolated from three plants viz. Allophylus serratus, Cissus quadrangularis and Vitex negundo were evaluated for their in vitro osteogenic activities. Primary cultures of osteoblasts were used to determine the effects of these components on osteoblast functions. Five (4, 6, 9, 12 and 14) of the fourteen compounds isolated led to increase in osteoblast differentiation and mineralization. These findings lend support to the use of Allophylus serratus, Cissus quadrangularis and Vitex negundo in traditional medicine.

Panax ginseng has anti-infective activity against opportunistic pathogen Pseudomonas aeruginosa by inhibiting quorum sensing, a bacterial communication process critical for establishing infection - Corrected Proof

2010-05-31

Abstract: Virulent factors produced by pathogens play an important role in the infectious process, which is regulated by a cell-to-cell communication mechanism called quorum sensing (QS). Pseudomonas aeruginosa is an important opportunistic human pathogen, which causes infections in patients with compromised immune systems and cystic fibrosis. The QS systems of P. aeruginosa use N-acylated homoserine lactone (AHL) as signal molecules. Previously we have demonstrated that Panax ginseng treatment allowed the animals with P. aeruginosa pneumonia to effectively clear the bacterial infection. We postulated that the ability to impact the outcome of infections is partly due to ginseng having direct effect on the production of P. aeruginosa virulence factors. The study explores the effect of ginseng on alginate, protease and AHL production. The effect of ginseng extracts on growth and expression of QS-controlled virulence factors on the prototypic P. aeruginosa PAO1 and its isogenic mucoid variant (PAOmucA22) was determined. Ginseng did not inhibit the growth of the bacteria, enhanced the extracellular protein production and stimulated the production of alginate. However, ginseng suppressed the production of LasA and LasB and down-regulated the synthesis of the AHL molecules. Ginseng has a negative effect on the QS system of P. aeruginosa, may explain the ginseng-dependent bacterial clearance from the animal lungs in vivo in our previous animal study. It is possible that enhancing and repressing activities of ginseng are mutually exclusive as it is a complex mixture, as shown with the HPLC analysis of the hot water extract. Though ginseng is a promising natural synergetic remedy, it is important to isolate and evaluate the ginseng compounds associated with the anti-QS activity.

Oligonol a low molecular weight polyphenol of lychee fruit extract inhibits proliferation of influenza virus by blocking reactive oxygen species-dependent ERK phosphorylation - Corrected Proof

2010-05-31

Abstract: The emergence of resistance to anti-influenza drugs calls for the search for new antiviral molecules with different resistance profiles. Polyphenolic compounds are found in various plants and have antiviral and antioxidative properties. We tested the hypothesis that oligonol, a lychee fruit-derived low molecular weight polyphenol, possesses anti-influenza effects by inhibiting phosphorylation of extracellular-signal-regulated kinases (ERK). Real time PCR, plaque assay, and immunofluorescence techniques were used to study the effects of oligonol on proliferation of influenza virus. Oligonol inhibits influenza virus proliferation by blocking attachment of the virus to MDCK cells and by suppression of nuclear export of influenza virus ribonucleoprotein (RNP). Infection of MDCK cells with influenza virus leads to an increase in production of reactive oxygen species (ROS) and induction of a ROS-dependent ERK phosphorylation. Inhibition of ERK activation by a dominant negative mutant of ERK or N-acetyl-cysteine (NAC) leads to inhibition of influenza RNP nuclear export. Phorbol 12-myristate 13-acetate (PMA) induces ROS production, ERK phosphorylation and enhances influenza proliferation in MDCK cells. Oligonol and NAC inhibit PMA-induced ERK phosphorylation and ROS production. Our studies suggest that the underlying mechanism for the inhibitory effect of oligonol on influenza virus RNP nuclear export is blocking of ROS-dependent induction of ERK phosphorylation.

The effects of Tanshinone IIA on blood–brain barrier and brain edema after transient middle cerebral artery occlusion in rats - Corrected Proof

Chao Tang, Hongli Xue, Changlin Bai, Rong Fu, Anhua Wu — 2010-05-31

Abstract: Disruption of blood–brain barrier (BBB) and edema formation play a key role in the development of neurological dysfunction after cerebral ischemia. In this study, the effects of Tanshinone IIA (Tan IIA), one of the active ingredients of Salvia miltiorrhiza root, on the BBB and brain edema after transient middle cerebral artery occlusion in rats were examined. Our study demonstrated that Tan IIA reduced brain infarct area, water content in the ischemic hemisphere. Furthermore, Tan IIA significantly decreased BBB permeability to Evans blue, suppressed the expression of intercellular adhesion molecule-1 (ICAM-1), matrix metalloproteinase-9 (MMP-9), inhibited the degradation of tight junction proteins zonula occludens-1 (ZO-1) and Occludin. These results demonstrated that Tan IIA was effective for attenuating the extent of brain edema formation in response to ischemia injury in rats, partly by Tan IIA's protective effect on the BBB. Our results may have implications in the treatment of brain edema in cerebral ischemia.

Proteomic analysis of anti-tumor effects by tetrandrine treatment in HepG2 cells - Corrected Proof

Zhixiang Cheng, Keming Wang, Jia Wei, Xiang Lu, Baorui Liu — 2010-05-31

Abstract: Tetrandrine (TET), a bis-benzylisoquinoline alkaloid isolated from the root of Hang-Fang-Chi (Stephenia tetrandra S Moore), exhibits broad pharmacological effects, including anti-tumor activity. Recently, the beneficial effects of TET on cytotoxicity towards tumor cells, radiosensitization, circumventing multidrug resistance, normal tissue radioprotection, and antiangiogenesis have been examined extensively. To explore the potential molecular mechanism of the anti-tumor effect of TET, we applied proteomic tools to profile the proteins in HepG2 cells subjected to TET treatment. The levels of 39 proteins in cells exposed to TET (IC50=5±0.6μg/ml) for 48h were observed to undergo significant alterations. Six proteins were identified by matrix assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS) using peptide fingerprinting from 10 protein spots (density difference >1.5-fold between the control and TET-treated group). Among them, 5 proteins were downregulated (proteasome activator complex subunit 3, 40S ribosomal protein S12, phosphoglycerate mutase 1, destrin, transaldolase) and 1 protein was upregulated (guanylate kinase 1) by TET treatment in HepG2 cells as determined by spot volume (P<0.05). Most of the identified proteins were associated with tumor growth, migration, and anti-tumor drug resistance. These data will be helpful in elucidating the molecular mechanism of TET's anti-tumor effect in HepG2 cells.

In vitro callus and in vivo leaf extract of Gymnema sylvestre stimulate β-cells regeneration and anti-diabetic activity in Wistar rats - Corrected Proof

A. Bakrudeen Ali Ahmed, A.S. Rao, M.V. Rao — 2010-05-31

Abstract: A methanol extract of Gymnema sylvestre leaf and callus showed anti-diabetic activities through regenerating β-cells. Optimum callus was developed under stress conditions of blue light with 2,4-D (1.5mg/l) and KN (0.5mg/l), which induced maximum biomass of green compact callus at 45 days, as determined by growth curve analysis. Leaf and optimum callus extracts contains gymnemic acid, which was analyzed using TLC, HPTLC and HPLC methods. The research reported here deals with leaf and callus extracts of G. sylvestre, which significantly increase the weight of the whole body, liver, pancreas and liver glycogen content in alloxan-induced diabetic rats (Wistar rats). The gymnemic acid of leaf and callus extracts significantly increases the regeneration of β-cells in treated rats, when compared with the standard diabetic rats. It could have potential as a pharmaceutical drug for insulin-dependent diabetes mellitus (IDDM).

Anticandidal activity of the essential oils of Thymus maroccanus and Thymus broussonetii and their synergism with amphotericin B and fluconazol - Corrected Proof

A. Saad, M. Fadli, M. Bouaziz, A. Benharref, N.-E. Mezrioui, L. Hassani — 2010-05-31

Abstract: The discovery of antifungal drugs had eradicated some infections that ravaged the humankind. But their indiscriminate use has led to the development of multidrug resistant pathogens. One strategy employed to overcome these resistance mechanisms is the use of combination of the essential oils (EOs) of medicinal plants and conventional drugs. In this study, we investigated a possible synergistic effect of the EOs of two Moroccan endemic thymes (Thymus broussonetii and T. maroccanus) with amphotericin B (Amp B) and fluconazol against Candida albicans. The fractional inhibitory concentration indices (FICI) of T. maroccanus and T. broussonetii EOs combined with Amp B and fluconazol, calculated from the checkerboard titer assay, were 0.49, 0.27, 0.37 and 0.3, respectively. Also, our results indicate that the synergistic effect of EOs with fluconazol was stronger than the combination with Amp B. All these data highlight that the EOs tested potentiate the antifungal action of Amp B and fluconazol, suggesting a possible utilization of these EOs in addition to antifungal drugs for the treatment of some candidiasis due to C. albicans. The use of these combinations is likely to reduce the minimum effective dose of the drugs, thus minimizing their toxic side effects and the treatment cost.

β-Ecdysone has bone protective but no estrogenic effects in ovariectomized rats - Corrected Proof

2010-05-31

Abstract: Estrogens exert beneficial effects in the bone. Their chronic use however bares several risks. Therefore intensive search for non-estrogenic, bone protective compounds is going on. We observed that an extract of Tinospora cordifolia has antiosteoporotic effects and identified 20-OH-Ecdysone (β-Ecdysone=Ecd) as a possible candidate for this action.Ovariectomized (ovx) rats were treated orally over 3 months with no Ecd (control) or 18, 57 or 121mg Ecd/day/animal. Estradiol-17β benzoate (E2) 159μg/day/animal) fed animals served as positive controls. Bone mineral density (BMD) of tibia was measured by quantitative computer tomography, serum Osteocalcin and CrossLaps were measured in a ligand binding assay. Utilizing an estrogen receptor (ER) containing cytosolic extract of porcine uteri the capability of Ecd to bind to ER was tested.Ecd did not bind to ER. BMD was reduced by more than 50% in the control. In the Ecd animals BMD was dose dependently higher. Serum CrossLaps was lower in the Ecd and E2 group while serum Osteocalcin levels were decreased in the E2 but increased in the Ecd fed animals.Ecd has an antiosteoporotic effect which does not involve activation of ER.

Anti-inflammatory effects of the willow bark extract STW 33-I (Proaktiv®) in LPS-activated human monocytes and differentiated macrophages - Corrected Proof

G.A. Bonaterra, E.U. Heinrich, O. Kelber, D. Weiser, J. Metz, R. Kinscherf — 2010-05-31

Abstract: Introduction: Willow bark extract is frequently used in the treatment of painful rheumatological diseases, such as arthritis and back pain. Its effect has been attributed to its main component salicin, but pharmacological studies have shown that the clinical efficacy of the willow bark extract cannot be explained by its salicin content alone. Therefore different modes of action have been suggested for the anti-inflammatory effect of willow bark extract. Here, we report in vitro data revelling the effect and mode of action of the aqueous willow bark extract STW 33-I as well as a water-soluble fraction (fraction E [Fr E]) in comparison with well-known non-steroidal anti-inflammatory drugs (NSAIDs) like aspirin (ASA) and diclofenac (Diclo) on pro-inflammatorily activated human monocytes and differentiated macrophages.Results: STW 33-I and the water-soluble Fr E showed concentration-dependent and significant anti-inflammatory effects in lipopolysaccharide-activated monocytes. Both inhibited the intracellular protein expression of tumour necrosis factor-alpha (TNFα) as well as the mRNA expression of TNFα and cyclooxygenase 2 (COX-2), and the release of nitric oxide (NO). In addition, apoptosis of pro-inflammatorily activated monocytes was induced. Furthermore, treatment of activated macrophages with STW 33-I inhibited the nuclear translocation of the p65 subunit of the nuclear transcription factor-kappa B (NF-κB p65).Conclusions: The present in vitro investigations suggest a significant anti-inflammatory activity of willow bark water extract STW 33-1 and of its water-soluble fraction by inhibiting pro-inflammatory cytokines (TNFα), COX-2 and nuclear translocation of the transcription factor NF-κB in pro-inflammatorily activated monocytes. Our results provide further evidence for the therapeutic use of STW 33-I in inflammation-related disorders.

Cinnamon extract regulates glucose transporter and insulin-signaling gene expression in mouse adipocytes - Corrected Proof

Heping Cao, Donald J. Graves, Richard A. Anderson — 2010-05-31

Abstract: Cinnamon extracts (CE) are reported to have beneficial effects on people with normal and impaired glucose tolerance, the metabolic syndrome, type 2 diabetes, and insulin resistance. However, clinical results are controversial. Molecular characterization of CE effects is limited. This study investigated the effects of CE on gene expression in cultured mouse adipocytes. Water-soluble CE was prepared from ground cinnamon (Cinnamomum burmannii). Quantitative real-time PCR was used to investigate CE effects on the expression of genes coding for adipokines, glucose transporter (GLUT) family, and insulin-signaling components in mouse 3T3-L1 adipocytes. CE (100μg/ml) increased GLUT1 mRNA levels 1.91±0.15, 4.39±0.78, and 6.98±2.18-fold of the control after 2-, 4-, and 16-h treatments, respectively. CE decreased the expression of further genes encoding insulin-signaling pathway proteins including GSK3B, IGF1R, IGF2R, and PIK3R1. This study indicates that CE regulates the expression of multiple genes in adipocytes and this regulation could contribute to the potential health benefits of CE.

Genistein aglycone: A new therapeutic approach to reduce endometrial hyperplasia - Corrected Proof

2010-05-31

Abstract: Objective: Endometrial hyperplasia without cytological atypia is commonly treated with progestins, but other treatment regimes may be available with equivalent efficacy and low side effects.Design: A randomized double-blind, placebo and progesterone-controlled clinical trial to evaluate the effects of genistein aglycone in reducing endometrial hyperplasia.Patients: A group of 56 premenopausal women with non-atypical endometrial hyperplasia were enrolled and received: genistein aglycone (n=19; 54mg/day); norethisterone acetate (n=19; 10mg/day on days 16–25 of the menstrual cycle) or placebo (n=18) for 6 months.Measurements: Hysteroscopy was performed with biopsies and symptomology assessed at baseline, 3 and 6 months of administration. The effect on estrogen (ER) and progesterone receptors (PR) expression in uterine biopsies were assessed after 3 and 6 months. For each treatment follicle stimulating hormone (FSH), luteinizing hormone (LH), estradiol (E2), sex hormone-binding globulin (SHBG) and progesterone (PG) levels were also evaluated.Results: After 6 months, 42% of genistein aglycone-administered subjects had a significant improvement of symptoms (histologically confirmed in the 29%) compared to 47% of norethisterone acetate subjects (histologically confirmed in the 31%), but only 12% in the placebo group with 19% exhibiting worsening symptoms and increased endometrial thickness. No significant differences were noted for hormone levels for any treatment, but immunohistochemical analysis revealed significantly reduced staining for ER-α and PR and enhanced ER-β1 staining in genistein-administered subjects associated with a complete regression of bleeding.Conclusions: These results suggest that genistein aglycone might be useful for the management of endometrial hyperplasia without atypia in women that cannot be treated with progestin.

Arylnaphthalene lignans from Taiwania cryptomerioides as novel blockers of voltage-gated K+ channels - Corrected Proof

2010-05-31

Abstract: Lignans are natural phytochemicals which exhibit multiple pharmacological effects such as anti-inflammation, antivirus and anti-tumor activities. Whether they have effects on neural tissues and ion channels is still unknown. The effects of several arylnaphathalene lignans purified from Taiwania cryptomerioides on voltage-gated K+ (Kv) channels in mouse neuroblastoma N2A cells were examined. These lignans included Taiwanin E, helioxanthin (HXT) and diphyllin. All lignans showed inhibitory effects on Kv channels and HXT was the most potent compound (IC50=1.7μM). The mechanism of HXT block was further investigated. Its action was found to be extracellular but not intracellular. HXT accelerated current decay, caused a left-shift in steady-state inactivation curve but had no effect on voltage-dependence of activation. HXT block was unaffected by intracellular K+ concentrations. Further, it did not affect ATP-sensitive K+ channels. Our data therefore suggest that HXT is a potent and specific blocker of Kv channels, possibly with an inhibitory mechanism involving acceleration of slow inactivation.

Icariin inhibits osteoclast differentiation and bone resorption by suppression of MAPKs/NF-κB regulated HIF-1α and PGE2 synthesis - Corrected Proof

Tsai-Pei Hsieh, Shiow-Yunn Sheu, Jui-Sheng Sun, Ming-Hong Chen — 2010-05-31

Abstract: Icariin has been reported to enhance bone healing and treat osteoporosis. In this study, we examined the detail molecular mechanisms of icariin on lipopolysaccharide (LPS)-induced osteolysis. Our hypothesis is that icariin can inhibit osteoclast differentiation and bone resorption by suppressing MAPKs/NF-κB regulated HIF-1α and PGE2 synthesis.After treatment with icariin, the activity of osteoclasts differentiation maker, tatrate resistances acid phosphatease (TRAP), significantly decreased at the concentration of 10−8M. Icariin (10−8M) reduced the size of LPS-induced osteoclasts formation, and diminished their TRAP and acid phosphatease (ACP) activity without inhibition of cell viability. Icariin also inhibited LPS-induced bone resorption and interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) expression. The gene expression of osteoprotegerin (OPG) was up-regulated, while receptor activator of NF-κB ligand (RANKL) was down-regulated. Icariin also inhibited the synthesis of cyclo-oxygenase type-2 (COX-2) and prostaglandin E2 (PGE2). In addition, icariin had a dominant repression effect on LPS-induced hypoxia inducible factor-1α (HIF-1α) expression of osteoclasts. On osteoclasts, icariin suppresses LPS-mediated activation of the p38 and JNK; while on the osteoblasts, icariin reduced the LPS-induced activation of ERK1/2 and I-kappa-B-alpha (IκBα), but increased the activation of p38.In conclusion, we demonstrated that icariin has an in vitro inhibitory effects on osteoclasts differentiation that can prevent inflammatory bone loss. Icariin inhibited LPS-induced osteoclastogenesis program by suppressing activation of the p38 and JNK pathway.

Ethanolic extracts of Brazilian red propolis promote adipocyte differentiation through PPARγ activation - Corrected Proof

2010-04-12

Abstract: Aim of the study: The aim of present study was to investigate the effects of ethanolic extracts of red propolis (EERP) on adipogenesis and evaluate the molecular basis for their anti-obesity effects.Materials and methods: We tested whether EERP alone could induce differentiation of 3T3-L1 cells, regulate the expression of adipocyte-specific genes and reverse inhibitory effects of TNF-α on their differentiation. Next, we performed a luciferase reporter gene assay to test whether EERP could enhance transcriptional activities of PPARγ and adiponectin promoter activities.Results: EERP strongly induced differentiation of 3T3-L1 preadipocytes into adipocytes, and enhanced the PPARγ transcriptional activity and adiponectin promoter activity. In addition, EERP attenuated the inhibitory effect of TNF-α on adipocyte differentiation and adiponectin production in mature adipocytes.Conclusion: The present study indicates that EERP enhance differentiation of 3T3-L1 adipocytes in part by its potency of PPARγ activation and are capable of reversing inhibitory effects of TNF-α on adipocyte differentiation and adiponectin expression. These results suggest the value of EERP as a diet supplement for prevention and treatment of obesity and obesity-associated disorders.

Passiflora incarnata L. (Passionflower) extracts elicit GABA currents in hippocampal neurons in vitro, and show anxiogenic and anticonvulsant effects in vivo, varying with extraction method - Corrected Proof

2010-04-12

Abstract: Potential mechanisms of Passiflora incarnata extracts and the effect of extraction methods on ingredients and biological effects were explored. Using the same batch of plant material, total flavonoid yields as measured by high-performance liquid chromatography coupled to diode array detection (HPLC–DAD) increased substantially with hot versus cold extraction methods.Whole Passiflora extract induced prominent, dose-dependent direct GABAA currents in hippocampal slices, but the expected modulation of synaptic GABAA currents was not seen. GABA was found to be a prominent ingredient of Passiflora extract, and GABA currents were absent when amino acids were removed from the extract.Five different extracts, prepared from a single batch of Passiflora incarnata, were administered to CF-1 mice for 1 week in their drinking water prior to evaluation of their behavioral effects. Anticonvulsant effects against PTZ-induced seizures were seen in mice that received 2 of the 5 Passiflora extracts. Instead of the anxiolytic effects described by others, anxiogenic effects in the elevated plus maze were seen in mice receiving any of the 5 Passiflora extracts.

Antimicrobial, antiviral and cytotoxic activity of extracts and constituents from Polygonum spectabile Mart. - Corrected Proof

2010-04-12

Abstract: Polygonum spectabile is used in Brazil for treatment of several infection diseases. Extracts and constituents isolated from this species were evaluated for cytotoxicity and effects on 15 bacterias and yeasts as well on 4 viruses strains (HHV-1, VACV-WR, EMCV, DEN-2). Less polar extracts were effective against Staphylococcus aureus, Bacillus subtillis, Micrococcus luteus, M. canis and Tricophyton mentagrophytes and T. rubrum. Two known chalcones and 3-O-β-d-glucosyl-β-sitosterol were isolated. The ethanol extract was the only one to show antiviral activity (CE50<30μg/ml). One chalcone has inhibited the growth of several bacteria and was significantly active against dermathophytes. The 3 compounds isolated have shown moderate cytotoxicity against Vero and LLCMK2 cells (CC50 <50μg/ml). These results support the use of P. spectabile as antimicrobial agent.

Juice of Bryophyllum pinnatum (Lam.) inhibits oxytocin-induced increase of the intracellular calcium concentration in human myometrial cells - Corrected Proof

2010-04-12

Abstract: The use of preparations from Bryophyllum pinnatum in tocolysis is supported by both clinical (retrospective comparative studies) and experimental (using uterus strips) evidence. We studied here the effect of B. pinnatum juice on the response of cultured human myometrial cells to stimulation by oxytocin, a hormone known to be involved in the control of uterine contractions by increasing the intracellular free calcium concentration ([Ca2+]i).In this work, [Ca2+]i was measured online during stimulation of human myometrial cells (hTERT-C3 and M11) with oxytocin, which had been pre-incubated in the absence or in the presence of B. pinnatum juice. Since no functional voltage-gated Ca2+ channels could be detected in these myometrial cells, the effect of B. pinnatum juice was as well studied in SH-SY5Y neuroblastoma cells, which are known to have such channels and can be depolarised with KCl.B. pinnatum juice prevented the oxytocin-induced increase in [Ca2+]i in hTERT-C3 human myometrial cells in a dose-dependent manner, achieving a ca. 80% inhibition at a 2% concentration. Comparable results were obtained with M11 human primary myometrial cells. In hTERT-C3 cells, prevention of the oxytocin-induced increase in [Ca2+]i was independent of the extracellular Ca2+ concentration and of voltage-dependent Ca2+-channels. B. pinnatum juice delayed, but did not prevent the depolarization-induced increase in [Ca2+]i in SH-SY5Y cells.Taken together, the data suggest a specific and concentration-dependent effect of B. pinnatum juice on the oxytocin signalling pathway, which seems to corroborate its use in tocolysis. Such a specific mechanism would explain the rare and minor side-effects in tocolysis with B. pinnatum as well as its high therapeutic index.

In vitro and in vivo antitumor activity of Macrothelypteris torresiana and its acute/subacute oral toxicity - Corrected Proof

2010-04-12

Abstract: The aim of this study was to evaluate the antitumor potential of Macrothelypteris torresiana by studying in vitro antitumor activity of the protoapigenone, as well as in vivo antitumor activity and acute/subacute oral toxicity of the total flavonoid fraction from the roots of M. torresiana. Considering that the protoapigenone is a main constituent of the total flavonoid fraction and it might play a key role in the antitumor activity of M. torresiana, the MTT assay was used to investigate the in vitro antitumor activity of the protoapigenone. Our study revealed that the protoapigenone of M. torresiana showed significant antitumor activity towards Hep G2, Tca-8113, MCF-7, M5 and K562 with IC50 values of 2.3, 0.6, 0.8, 0.3 and 0.9μg/ml, respectively. The antitumor potential of the total flavonoid fraction was evaluated using preparations 1, 2 and 3, which were prepared by total flavonoid fraction directly diluted with sterile saline, dissolved using sodium carboxymethyl cellulose (CMC-Na) and included by hydroxypropyl-β-cyclodextrin, respectively. These were investigated in vivo using mouse sarcoma S-180 in BALB/c mice after completing tumor inoculation for 24h. Pronounced antitumor activity was observed in the treated groups for preparations 2 and 3, and the high and medium doses in particular showed very high inhibition ratio of tumor growth (>50%). No significant difference was observed when compared to the positive control group (5-fluorouracil). The acute/subacute oral toxicity test was performed, and the results of acute oral toxicity showed that the LD50 values of preparations 2 and 3 were 2.76 and 0.87g/kg body wt., respectively. According to the results of the subacute oral toxicity study, the total flavonoid fraction had low toxicity. The overall results of this study suggest that the total flavonoid fraction from the roots of M. torresiana shows significant antitumor activity and represents a potential source of medicine for the treatment of cancer.