Phytomedicine - Articles in Press

Effect of purified saponin mixture from Astragalus corniculatus on enzyme- and non-enzyme-induced lipid peroxidation in liver microsomes from spontaneously hypertensive rats and normotensive rats - Corrected Proof

2010-02-03

Abstract: The aim of the following study was to evaluate the effect of a purified saponin mixture (PSM), isolated from Astragalus corniculatus Bieb. (Fabaceae), on enzyme-induced and non-enzyme-induced lipid peroxidation (LPO), in liver microsomes from spontaneously hypertensive rats (SHRs) – strain Okamoto Aoki, as compared to normotensive Wistar rats (NTRs).The enzyme-induced lipid peroxidation was performed by incubating rat liver microsomes with carbonetetrachloride (CCl4) in the presence of NADPH. In nonenzyme-induced LPO, the microsomes were incubated with a solution of iron sulphate and ascorbinic acid (Fe2+/AA). The effect of PSM (196.5μg/ml) was assessed at 20 minutes’ incubation time. MDA, a product of LPO, was measured spectrophotometrically.The results of our study showed that the initial MDA quantity in SHRs was significantly higher, than in NTRs. The incubation of the microsomes from both strains with PSM (196.5μg/ml), resulted in significant reduction of MDA level, by 25% in SHRs. In NTRs, the formation of MDA was unchanged.In enzyme-induced LPO model, PSM significantly decreased the formation of MDA, by 55% in NTRs and by 35% in SHRs, compared to the respective control groups. In the model of non-enzyme induced LPO, PSM significantly decreased the formation of MDA by 95% in NTRs and practically restored it to the control level. The MDA quantity in SHR's microsomes was reduced by 25%.According to the results of this experiment we could conclude that PSM, isolated from Astragalus corniculatus, shows antioxidant activity both in SHRs and NTRs and the effect in NTRs is more pronounced.

Selective induction of apoptosis in glioma tumour cells by a Gynostemma pentaphyllum extract - Corrected Proof

L. Schild, B.H. Chen, P. Makarov, K. Kattengell, K. Heinitz, G. Keilhoff — 2010-01-27

Abstract: At low concentration H2O2 is an important signal molecule in proliferation of tumour cells. We report about a study investigating the effect of an ethanolic extract from Gynostemma pentaphyllum on proliferation of C6 glioma tumour cells and cellular H2O2 concentration. The proliferation of these cells was maximal at about 1 μM extracellular H2O2. HPLC-finger prints of the extract revealed a set of saponines as essential components. In C6 gioma cells the extract caused increase in super oxide dismutase (SOD) activity, in the amount of SOD protein, and in cellular H2O2 concentration. It inhibited cell proliferation and induced activation of caspase 3 as indication of apoptosis. No effect of the extract was observed on the proliferation of astrocytes of a primary cell culture. From these findings we suggest that the ethanolic extract from Gynostemma pentaphyllum may selectively shift the H2O2 concentration to toxic levels exclusively in tumour cells due to increased SOD activity. It may have a high potency in cancer therapy and cancer prophylaxis.

Synergistic effects of β-aescin and 5-fluorouracil in human hepatocellular carcinoma SMMC-7721 cells - Corrected Proof

Z.J. Ming, Y. Hu, Y.H. Qiu, L. Cao, X.G. Zhang — 2010-01-27

Abstract: The effects and mechanisms of action of β-aescin and 5-fluorouracil (5-FU), alone and in combination, were studied in human hepatocellular carcinoma SMMC-7721 cells. Growth inhibition, cell cycle distribution, apoptosis, Bcl-2 expression and caspase activity were assessed. The Isobole-method/interaction-index analysis was applied to evaluate the synergy, additivity or antagonism of these agents. The results indicate that mixtures of β-aescin and 5-FU showed a synergistic effect on the 50% inhibitory effect when their ratio was 4:1 when compared with either agent alone. The mechanism of action could be through the synergistic arrest of the cell cycle, induction of apoptosis, activation of caspases-3, 8 and 9, and down-regulation Bcl-2 expression. The results suggest that mixtures of these two agents had a synergistic inhibitory effect on SMMC-7721 cells, an observation which might be useful for the further development of anti-cancer drugs.

Mechanism of the vasodilator effect of Euxanthone in rat small mesenteric arteries - Corrected Proof

D.V. Câmara, V.S. Lemos, M.H. Santos, T.J. Nagem, S.F. Cortes — 2010-01-25

Abstract: In the present work we investigated the mechanism involved in the vasodilator effect induced by euxanthone in rat small mesenteric arteries. We observed that euxanthone induced concentration-dependent vasodilatation in arteries by a mechanism independent on the release of endothelial factors, such as nitric oxide (NO) and cyclooxygenase-derived factors. In addition our results also suggest that euxanthone induced its vasodilator effect through inhibition of calcium-sensitive mechanisms activated by protein kinase C, rather than by inhibition of contractions dependent on the release of the intracellular calcium stores or by inhibition of voltage-operated calcium channels.

Anti-proliferative effects of carvacrol on a human metastatic breast cancer cell line, MDA-MB 231 - Corrected Proof

K.M. Arunasree — 2010-01-25

Abstract: Purpose: Although the anti-tumor effects of carvacrol have been demonstrated earlier, the exact underlying molecular mechanisms involved in its action have not been defined and in the present study an attempt has been made to identify the mechanism of carvacrol induced cell death in human metastatic breast cancer cells, MDA-MB 231.Methods: Apoptosis induced by carvacrol was determined based on different assays like MTT assay, Annexin V, mitochondrial membrane potential assay, multicaspase activation assay and cell cycle analysis by flow cytometer. Cleavage of PARP, cytochrome c release and modulation of Bax and Bcl2 ratio by Western blot analysis were also studied.Results: The study clearly showed induction of apoptosis by carvacrol in MDA-MB 231 cells dose dependently at an IC50 of 100μM with a decrease in the mitochondrial membrane potential of the cells resulting in release of cytochrome c from mitochondria, caspase activation and cleavage of PARP.Conclusion: The data in the present study clearly demonstrated anti-tumor effects of carvacrol on human metastatic breast cancer cells, MDA-MB 231, and that the compound could have a potential therapeutic significance in treating cancer.

Apoptosis of human tumor cell lines by a lectin (futalin) of Artocarpus incisa seeds - Corrected Proof

Maria do Carmo Avides, J.A. Teixeira, A. Vicente — 2010-01-22

Abstract: A galactose-binding lectin (frutalin) from the seeds of Artocarpus incisa was extracted and purified by affinity chromatography on galactomanan columns. Human cervical carcinoma (HeLa) and human mammary carcinoma (MCF7) cell lines were used to check the effects of frutalin on cell proliferation and apoptosis. Maximum growth inhibition (98%) was observed with MCF7 cells, followed by HeLa cells (92%). Non-transformed human umbilical cord cells, HUVEC, were not affected. Morphological observations showed that frutalin-treated HeLa and MCF-7 cells displayed apoptotic characteristics such as nuclear fragmentation and appearance of membrane-enclosed apoptotic bodies. Apoptosis of HeLa and MCF7 cells was confirmed by immunostaining for active caspase 3. We reason that the results presented therein may lead to the possible use of frutalin in cancer therapy.

The novel anti-hyperglycemic effect of Paeoniae radix via the transcriptional suppression of phosphoenopyruvate carboxykinase (PEPCK) - Corrected Proof

2010-01-22

Abstract: The antidiabetic actions of Paeoniae Radix involve stimulating glucose uptake and reducing glucose absorption. However, the importance of this herb in the transcriptional regulation of hepatic gluconeogenesis has not previously been investigated, although hepatic gluconeogenesis contributes the most to fasting hyperglycemia. Using rats with streptozotocin-induced diabetes and db/db mice, the dose- and time-dependent suppressive effects of the ethanol extract of Paeoniae Radix (PR-Et) on diabetic hyperglycemia and phosphoenopyruvate carboxykinase (PEPCK) transcription are first demonstrated. Second, by employing H4IIE cells, the inhibitory action of PR-Et on both dexamethasone- and 8-bromo-cAMP-induced-PEPCK expression was also confirmed without causing any cytotoxicity. In addition, this inhibitory effect could be sustained for over 24 h with repeated treatment. Most importantly, PR-Et's action was unaffected by either insulin desensitization or palmitate stimulation. Finally, paeonol and paeoniflorin, two well-known constituents in Paeoniae Radix, did not suppress PEPCK expression at testing concentration. In conclusion, it was clearly demonstrated that transcriptional inhibition of gluconeogenesis is one of the important antidiabetic actions of Paeoniae Radix. Future development of this herb as a dietary supplement or drug should bring substantial benefits for the diabetic population.

The pharmacokinetics of C-glycosyl flavones of Hawthorn leaf flavonoids in rat after single dose oral administration - Corrected Proof

L.Y. Ma, R.H. Liu, X.D. Xu, M.Q. Yu, Q. Zhang, H.L. Liu — 2010-01-22

Abstract: Hawthorn leaf flavonoids (HLF) are used in the treatment of cardiovascular diseases. Various potential pharmacodynamic effects have been observed for vitexin-4″-O-glucoside (VOG) and vitexin-2″-O-rhamnoside (VOR) which are the main constituents of HLF. The aim of this study was to investigate the pharmacokinetics of VOG and VOR when a single dose of HLF was administrated orally. The levels of VOG and VOR in plasma, tissues (heart, liver, spleen, lung, kidney and brain), bile, urine and feces were measured by HPLC-UV. The results showed that VOG and VOR have the similar pharmacokinetics. Both of them were absorbed quickly into plasma with maximal plasma concentrations of VOG and VOR being reached within 0.75h. The mean elimination half-life (t1/2) of VOG and VOR were 2.53h and 2.32h, respectively. High levels of tissue distribution of VOG and VOR were observed in liver and kidney. No VOG and VOR were detected in brain tissue. There was no long-term accumulation of VOG and VOR in rat tissues examined. The total recovery of the dose in 24 hours was 64.91% (0.70% in urine; 64.21% in feces) for VOG and 89.01% (0.72% in urine; 88.29% in feces) for VOR. The cumulative VOG and VOR excreted in bile represented 0.58% and 13.38% of the doses, respectively. VOG and VOR in HLF were not efficiently absorbed in the rodent gastrointestinal tract.

A standardized aqueous extract of Anoectochilus formosanus modulated airway hyperresponsiveness in an OVA-inhaled murine model - Corrected Proof

C.-C. Hsieh, H.-B. Hsiao, W.-C. Lin — 2010-01-22

Abstract: Anoectochilus formosanus HAYATA, a Chinese herb, is a valued folk medicine for fever, pain, and diseases of the lung and liver. Allergic asthma is characterized by increased serum IgE level and inflammation of the airways with high levels of interleukin (IL)-4 and IL-5 in bronchoalveolar lavage fluids (BALF). Constriction of airway smooth muscle and development of airway hyperresponsiveness (AHR) are the most important symptoms of allergic asthma. In our previous study, a standardized aqueous extract of A. formosanus (SAEAF) was used to modulate innate immunity of normal mice. In this study, airway inflammatory infiltrations, including T cell differentiation, cytokine modulation, allergic antibodies estimation, pulmonary pathology, and enhanced pause (Penh) of AHR were used to evaluate SAEAF treatment of an ovalbumin (OVA)-inhaled airway allergic murine model. The resulting cytokine profiles demonstrated that SAEAF can significantly reduce Th2 polarization after administration of SAEAF in OVA inhalation. These results also suggest that SAEAF modulates cytokine secretion in allergic asthma. Modulated natural T regulatory cells (CD25+/CD4+, Treg) were also shown to increase immuno-suppression in the allergic lung inflammation and further down-regulate airway inflammatory infiltration in eosinophils and macrophages. Finally, decreased airway anti-OVA IgE secretion and reduced AHR were observed. Our results indicate that the administration of SAEAF can modulate cytokines and T cell subpopulation by regulating inflammatory cell infiltration and modulating the allergic response.

Corrected Proof

Jenny J. Zhou — 2010-01-21

Plant foods are prominent features of a healthy diet. They include fruits, vegetables, legumes, whole grains, and nuts. In addition to providing energy and essential vitamins and minerals, plant foods contribute thousands of phytochemicals to the human diet. Although there is ample evidence that diets rich in plant foods are beneficial, evidence that these benefits are due to specific phytochemicals are more limited. “An Evidence-Based Approach to Dietary Phytochemicals” provides a concise synthesis of the basic scientific, observational, clinical data now available and their applications to health promotion and treatment.

The plant extract Isatis tinctoria L. extract (ITE) inhibits allergen-induced airway inflammation and hyperreactivity in mice - Corrected Proof

A. Brattström, A. Schapowal, M.A. Kamal, I. Maillet, B. Ryffel, R. Moser — 2010-01-21

Abstract: Background: The herbal Isatis tinctoria extract (ITE) inhibits the inducible isoform of cyclooxygenase (COX-2) as well as lipoxygenase (5-LOX) and therefore possesses anti-inflammatory properties. The extract might also be useful in allergic airway diseases which are characterized by chronic inflammation.Methods: ITE obtained from leaves by supercritical carbon dioxide extraction was investigated in ovalbumin (OVA) immunised BALB/c mice given intranasally together with antigen challenge in the murine model of allergic airway disease (asthma) with the analysis of the inflammatory and immune parameters in the lung.Results: ITE given with the antigen challenge inhibited in a dose related manner the allergic response. ITE diminished airway hyperresponsiveness (AHR) and eosinophil recruitment into the bronchoalveolar lavage (BAL) fluid upon allergen challenge, but had no effect in the saline control mice. Eosinophil recruitment was further assessed in the lung by eosinophil peroxidase (EPO) activity at a dose of 30μg ITE per mouse. Microscopic investigations revealed less inflammation, eosinophil recruitment and mucus hyperproduction in the lung in a dose related manner. Diminution of AHR and inflammation was associated with reduced IL-4, IL-5, and RANTES production in the BAL fluid at the 30μg ITE dose, while OVA specific IgE and eotaxin serum levels remained unchanged.Conclusion: ITE, which has been reported inhibiting COX-2 and 5-LOX, reduced allergic airway inflammation and AHR by inhibiting the production of the Th2 cytokines IL-4 and IL-5, and RANTES.

Inhibition of cytochrome P450 3A4 activity by schisandrol A and gomisin A isolated from Fructus Schisandrae chinensis - Corrected Proof

C.-K. Wan, A.K. Tse, Z.-L. Yu, G.-Y. Zhu, H. Wang, D.W.F. Fong — 2010-01-21

Abstract: We studied the effects of schisandrol A (SCH) and gomisin A (GOM), two of the main bioactive components of Fructus Schisandrae chinensis, on cytochrome P450-3A4 (CYP3A4) activity and cellular glutathione (GSH) level. In a cell-free system both SCH and GOM inhibited CYP3A4 activity with IC50 values of 32.02μM and 1.39μM, respectively. SCH or GOM at concentrations up to 100μM did not alter cellular GSH level in regular HepG2 cells and P-glycoprotein overexpressing HepG2-DR cells. Since SCH and GOM may reverse multidrug resistance (MDR) by impeding the activity of P-glycoprotein, a membrane xenobiotic exporter, SCH or GOM could affect cellular drug metabolism in addition to drug uptake.

Perezone and its isomer isoperezone induce caspase-dependent and caspase-independent cell death - Corrected Proof

2010-01-21

Abstract: In this study, we investigated the cytotoxic effect of perezone, a constituent isolated from the roots of Perezia spp. and of its synthetic isomer isoperezone on the K562 human leukemia cell line. Perezone showed greater cytotoxic effect than isoperezone but both compounds were found to induce cytotoxicity trough a caspase-dependent and a caspase-independent mechanisms; important changes in their light scattering properties, phosphatidylserine translocation and mitochondrial membrane potential disruption were detected by cytometry. The mechanism of death induction of each compound showed interesting concentration-dependent differences. Neither compound induced the apoptosis inducing factor.

Effects of triterpenes from Ganoderma lucidum on protein expression profile of HeLa cells - Corrected Proof

2010-01-21

Abstract: To elucidate the cytotoxicity mechanism of Garnoderma triterpenes, a chemoproteomic study using five purified ganoderic acids, ganoderic acid F (GAF), ganoderic acid K (GAK), ganoderic B (GAB), ganoderic acid D (GAD) and ganoderic acid AM1 (GAAM1) was conducted. GAF, GAK, GAB, GAD and GAAM1 treatment for 48 h inhibited the proliferation of HeLa human cervical carcinoma cells with IC50 values of 19.5±0.6μM, 15.1±0.5μM, 20.3±0.4μM, 17.3±0.3μM, 19.8±0.7μM, respectively. The protein expression profiles of HeLa cells treated with each ganoderic acid at dose of 15μM for 48h were checked using two-dimensional electrophoresis (2-DE). The possible target-related proteins of ganoderic acids, i.e. proteins with same change tendency in all five ganoderic acids-treated groups compared with control, were identified using MALDI-TOF MS/MS. Twelve proteins including human interleukin-17E, eukaryotic translation initiation factor 5A (eIF5A), peroxiredoxin 2, ubiquilin 2, Cu/Zn-superoxide dismutase, 14-3-3 beta/alpha, TPM4-ALK fusion oncoprotein type 2, PP2A subunit A PR65-alpha isoform, nucleobindin-1, heterogeneous nuclear ribonucleoprotein K, reticulocalbin 1 and chain A of DJ-1 protein were identified. Ganoderic acids might exert their cytotoxicity by altering proteins involved in cell proliferation and/or cell death, carcinogenosis, oxidative stress, calcium signaling and ER stress.

A novel polysaccharide, isolated from Angelica sinensis (Oliv.) Diels induces the apoptosis of cervical cancer HeLa cells through an intrinsic apoptotic pathway - Corrected Proof

W. Cao, X.-Q. Li, X. Wang, H.-T. Fan, X.-N. Zhang, Y. Hou, S.-B. Liu, Q.-B. Mei — 2010-01-21

Abstract: A novel polysaccharide isolated from Angelica sinensis, named APS-1d showed cytotoxic activity towards several cancer cell lines in vitro. However, the precise antitumor mechanisms of this compound are unknown. In this study, we investigated the pro-apoptotic effects of APS-1d in human cervical cancer HeLa cells both in vitro and in vivo, and further elucidated the mechanisms of this action. Inhibition of HeLa cell proliferation was determined by MTT assay and the therapeutic efficacy of APS-1d was evaluated by human cancer xenografts in nude mice. Cell apoptosis was examined with flow cytometry and TUNEL assay. The mechanism of action of APS-1d was investigated by Western blot analysis. APS-1d decreased HeLa cell proliferation in a concentration- and time-dependent manner in vitro. In addition, APS-1d significantly inhibited tumor growth in athymic nude mice. Characteristic manifestations of apoptosis including apoptotic morphological features and the sub- G0/G1 peaks were observed when the cells were treated with APS-1d. Further analysis showed that APS-1d-induced apoptosis was associated with the regulation of Bcl-2 family protein expression, a decrease in the mitochondrial membrane potential, and an increase in the cytosolic cytochrome c levels. Sequentially, APS-1d increased the activities of caspase-9, -3, and poly (ADP-ribose) polymerase in a concentration-dependent manner, however, no obvious activation of Bid and caspase-8 was observed. Pretreatment with Z-LEHD-FMK, a specific inhibitor of caspase-9, significantly attenuated APS-1d-induced cell apoptosis, and activation of caspase-3. Taken together, our studies indicate that APS-1d is capable of inhibiting HeLa cell proliferation and inducing apoptosis in these cells which primarily involves the activation of the intrinsic mitochondrial pathway.

Valeriana officinalis root extracts have potent anxiolytic effects in laboratory rats - Corrected Proof

K. Murphy, Z.J. Kubin, J.N. Shepherd, R.H. Ettinger — 2009-12-30

Abstract: Valerian root (Valeriana officinalis) is a popular and widely available herbal supplement, primarily used to treat insomnia and anxiety. Until recently, its mechanism of action has remained unknown. Neurobiological research has begun to show that the herb, with its active valerenic acid, interacts with the GABAA-ergic system, a mechanism of action similar to the benzodiazepine drugs. This series of experiments sought to corroborate these findings with behavioral measures, compare them to the benzodiazepine diazepam, and to analyze the chemical composition of Valeriana officinalis. Rats were administered either ethanol (1ml/kg), diazepam (1mg/kg), valerian root extract (3ml/kg), valerenic acid (3mg/kg), or a solution of valerenic acid and exogenous GABA (75μg/kg and 3.6μg/kg, respectively) and assessed for the number of entries and time spent on the open arms of an elevated plus maze. Results showed that there was a significant reduction in anxious behavior when valerian extract or valerenic acid exposed subjects were compared to the ethanol control group. The evidence supports Valeriana officinalis as a potential alternative to the traditional anxiolytics as measured by the elevated plus maze.

Osthol regulates hepatic PPARα-mediated lipogenic gene expression in alcoholic fatty liver murine - Corrected Proof

Fan Sun, Mei-lin Xie, Jie Xue, Heng-bin Wang — 2009-12-30

Abstract: Our previous studies found that osthol, an active constituent isolated from Cnidium monnieri (L.) Cusson (Apiaceae), could ameliorate the accumulation of lipids and decrease the lipid levels in serum and hepatic tissue in alcohol-induced fatty liver mice and rats. The objective of this study was to investigate its possible mechanism of the lipid-lowering effect. A mouse model with alcoholic fatty liver was induced by orally feeding 52% erguotou wine by gavage when they were simultaneously treated with osthol 10, 20, 40mg/kg for 4 weeks. The BRL cells (rat hepatocyte line) were cultured and treated with osthol at 25, 50, 100, 200μg/ml for 24h. The mRNA expressions of peroxisome proliferator-activated receptor (PPAR) α, diacylglycerol acyltransferase (DGAT), 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) reductase and cholesterol 7α-hydroxylase (CYP7A) in mouse hepatic tissue or cultured hepatocytes were determined by reverse transcription polymerase chain reaction (RT-PCR). After treatment with osthol, the PPARα mRNA expression in mouse liver and cultured hepatocytes was increased in dose dependent manner, while its related target genes for mRNA expression, e.g., DGAT and HMG-CoA reductase, were decreased, the CYP7A was inversely increased. And osthol-regulated mRNA expressions of DGAT, HMG-CoA reductase and CYP7A in the cultured hepatocytes were abrogated after pretreatment with specific inhibitor of PPARα, MK886. It was concluded that osthol might regulate the gene expressions of DGAT, HMG-CoA reductase and CYP7A via increasing the PPARα mRNA expression.

Evaluation of bone quality and quantity in osteoporotic mice – The effects of genistein and equol - Corrected Proof

2009-12-28

Abstract: The technology of gene manipulation is often used in mice. A crucial point for osteoporosis research is the evaluation of biomechanical and morphologic parameters. These parameters, however, are difficult to measure in mice. Nevertheless, this study demonstrates the capability of using techniques for the evaluation of bone quality and quantity after various treatments in osteopenic mice.After ovariectomy, 60 C57BL/6J mice were divided into 4 groups and were fed a soy-free diet (C) supplemented with estradiol, genistein or equol for 3 months. To analyze the osteoprotective effects of the tested supplements, we evaluated the bone biomechanical properties, histomorphometric changes and bone mineral density of the proximal tibiae metaphysis.The biomechanical parameters of genistein (GEN) were shown to be similar to those levels observed with estradiol (E). The biomechanical parameters of both GEN and E were significantly superior to those observed with C. Supplementation with equol (EQO) demonstrated higher mean biomechanical values than those observed with C. The histomorphometric evaluation demonstrated an increased number of nodes in mice treated with GEN and E as compared to the mice treated with EQO and C. Treatment with E and EQO led to improved cortical bone, which was only partly seen with the mice treated with GEN. The analysis of the bone mineral density (BMD) demonstrated that treatment with GEN and E resulted in a significant improvement as compared to the mice treated with C, while the cancellous density was significantly increased in all of the supplementation groups.This study conclusively demonstrated that bone quality and quantity parameters can be measured in mice. Furthermore, biomechanical and morphologic evaluations were shown to be reliable for use in mice. Further studies may combine these techniques with gene manipulation technology to better understand osteoporosis. Treatment with GEN resulted in improved biomechanical results and enhancement of morphologic parameters.

An aqueous extract of Ammi visnaga fruits and its constituents khellin and visnagin prevent cell damage caused by oxalate in renal epithelial cells - Corrected Proof

P. Vanachayangkul, K. Byer, S. Khan, V. Butterweck — 2009-12-28

Abstract: Teas prepared from the fruits of Ammi visnaga L. (syn. “Khella”) have been traditionally used in Egypt as a remedy to treat kidney stones. It was the aim of our study to evaluate the effect of a Khella extract (KE) as well as the two major constituents khellin and visnagin on renal epithelial injury using LLC-PK1 and Madin-Darby-canine kidney (MDCK) cells. Both cell lines provide suitable model systems to study cellular processes that are possibly involved in the development of a renal stone. LLC-PK1 and MDCK cell lines were exposed to 300μM oxalate (Ox) or 133μg/cm2 calcium oxalate monohydrate (COM) in presence or absence of 10, 50, 100 or 200μg/mL KE. To evaluate cell damage, cell viability was assessed by determining the release of lactate dehydrogenase (LDH). KE (e.g. 100μg/ml) significantly decreased LDH release from LLC-PK1 (Ox: 8.46+0.76%; Ox + 100μg/ml KE: 5.41+0.94%, p<0.001) as well as MDCK cells (Ox: 30.9+6.58%; Ox+100μg/ml KE: 17.5+2.50%, p<0.001), which indicated a prevention of cell damage. Similar effects for KE were observed in both cell lines when COM crystals were added. In LLC-PK1 cells khellin and visnagin both decreased the % LDH release significantly in cells that were pretreated with Ox or COM crystals. However, khellin and visnagin exhibited different responses in MDCK cells. Whereas khellin slightly reduced the % LDH release after exposure of the cells to Ox and COM crystals, visnagin significantly decreased % LDH release only after COM crystal exposure. Overall both compounds were more active in LLC-PK1 than in MDCK cells. In summary, exposure of renal epithelial cells to Ox or COM crystals was associated with a significant release of LDH indicating cell injury. Our data demonstrate that KE as well as khellin and visnagin could prevent renal epithelial cell damage caused by Ox and COM and could therefore play a potential role in the prevention of stone formation associated with hyperoxaluria.

Bactericidal and anti-inflammatory properties of a standardized Echinacea extract (Echinaforce®): Dual actions against respiratory bacteria - Corrected Proof

S.M. Sharma, M. Anderson, S.R. Schoop, J.B. Hudson — 2009-12-28

Abstract: Common symptoms of upper respiratory infections, such as sore throat, cough, and inflammation, are often caused by bacteria, sometimes as a complication of virus infection. Extracts of Echinacea purpurea (Asteraceae) have been advocated traditionally for use by individuals suffering from these symptoms, although the underlying basis for the beneficial effects of Echinacea is not known. We hypothesized that Echinacea could inactivate certain respiratory bacteria and could also reverse inflammatory effects caused by these bacteria in epithelial cells. In order to test this we used a commercial standardized extract of Echinacea purpurea (Echinaforce®), and a novel cytokine array system designed to measure simultaneously the levels of 20 different cytokines secreted by bronchial epithelial cell cultures in response to infection. Streptococcus pyogenes (Group A Strep), which is often associated with sore throat and more severe pulmonary infections, was readily inactivated by Echinacea, which also completely reversed the cellular pro-inflammatory response. Hemophilus influenzae and Legionella pneumophila were also readily inactivated, and their pro-inflammatory responses reversed. Staphylococcus aureus (methicillin-resistant and sensitive strains) and Mycobacterium smegmatis were less sensitive to the bactericidal effects of Echinacea however, but their pro-inflammatory responses were still completely reversed. In contrast some other pathogens tested, including Candida albicans, were relatively resistant. Thus Echinaforce® exerts a dual action against several important respiratory bacteria, a killing effect and an anti-inflammatory effect. These results support the concept of using a standardized Echinacea preparation to control symptoms associated with bacterial respiratory infections.

Investigations into the specific effects of rosemary oil at the receptor level - Corrected Proof

P. Sagorchev, J. Lukanov, A.M. Beer — 2009-12-25

Abstract: Rosemary oil is used frequently in phytotherapy. The objective of the present study was to investigate the extent to which rosemary oil shows other effects on the smooth muscles than the familiar spasmolytic effects. The effects of rosemary oil on the spontaneous contractile activity were investigated in in vitro experiments with circular smooth-muscle strips of the guinea pig stomach. Rosemary oil was found to have agonistic effects on the α1 and α2 adrenergic receptors. These effects can be registered at concentrations up to 25μl/l of rosemary oil. At higher concentrations the spasmolytic effect described in other reports could be detected. At concentrations above 100μl/l rosemary oil, the effect of 10−5M ACH is completely suppressed. The results permit the assumption that, besides the spasmolytic effects investigated to date, owing to its specific effects on the α2 adrenergic receptors of the nerve cells, rosemary oil brings about an additional improvement of local blood circulation and alleviates pain.

Effect of isoquinoline alkaloids from two Hippeastrum species on in vitro acetylcholinesterase activity - Corrected Proof

2009-12-07

Abstract: The treatment of neurological disorders and neurodegenerative diseases is related to the levels of acetylcholine (ACh) through the inhibition of acetylcholinesterase (AChE). Galanthamine, an important alkaloid isolated from the Amaryllidaceae family, is approved for the pharmacological treatment of Alzheimer's disease (AD) and acts by inhibiting the acetylcholinesterase (AChE) activity. In the present study, Ellman's method was used to verify the inhibition of AChE activity of some isoquinolines alkaloids such as galanthamine, montanine, hippeastrine and pretazettine. At the concentrations 1mM, 500μm and 100μm, galanthamine presented an AChE inhibition higher than 90%. Montanine inhibited, in a dose-dependent manner, more than 50% of the enzyme at 1mM concentration. With the concentrations 500μm and 100μm, 30-45% of AChE activity inhibition was detected. The alkaloids hippeastrine and pretazettine presented no significant inhibition of the AChE activity. The results demonstrate that montanine significantly inhibits AChE activity at the tested concentrations, suggesting the necessity of further investigations on this alkaloid use in treating neurological disorders.

Pharmacological effects of Catharanthus roseus root alkaloids in acetylcholinesterase inhibition and cholinergic neurotransmission - Corrected Proof

2009-12-07

Abstract: The leaves of Catharanthus roseus constitute the only source of the well known indolomonoterpenic alkaloids vincristine and vinblastine. In this work we studied the biological potential of the roots, which are used in several countries as decocts or hot water extracts for the treatment of a number of conditions. The aqueous extract strongly inhibited acetylcholinesterase (AchE) in an in vitro microassay, an effect ascribable mainly to serpentine (IC50 = 0.775μM vs physostigmine IC50 = 6.45μM) as assessed with the pure compound. Pure alkaloids were tested for muscarinic and nicotinic antagonism using rat ex-vivo preparations, namely, ileum and diaphragm/phrenic-nerve, respectively. Serpentine competitively blocked muscarinic receptors with a pA2 of 5.2, whereas the precursor ajmalicine up to 80μM was undistinguishable from control, and catharanthine exhibited an unsurmountable muscarinic antagonism at greater than 10μM concentrations. Nicotinic receptor mediated diaphragm contractions were fully inhibited by catharanthine (IC50 = 59.6μM) and ajmalicine (IC50 = 72.3μM), in a reversible but non-competitive manner, unlike the more potent nicotinic antagonist tubocurarine (IC50 = 0.35μM) whose competitive blockade was overcome by a physostigmine-induced increase in acetylcholine. Serpentine up to 100μM did not change diaphragm contractions suggesting reduced affinity for neuromuscular nicotinic receptors. Despite strong in vitro AchE inhibition, serpentine failed to restore diaphragm contractions upon submaximal tubocurarine blockade, suggesting that poor tissue penetration may prevent serpentine from inhibiting AchE in deep neuromuscular synapses in the ex-vivo preparation. To our knowledge, the present study is the first to assess the effect of C. roseus root extracts, as well as of serpentine, ajmalicine and catharanthine on AchE. The results described herein suggest that the currently overlooked C. roseus roots may constitute a promising source of compounds with pharmaceutical interest. Moreover, given serpentine's potent in vitro AchE inhibitory activity and low cholinergic receptor affinity, it is conceivable that minor structural modifications may yield a potent and selective AchE inhibitor, potentially useful for the pharmacological management of conditions such as Alzheimer's disease and/or myasthenia gravis.

Microcalorimetric assay on the antimicrobial property of five hydroxyanthraquinone derivatives in rhubarb (Rheum palmatum L.) to Bifidobacterium adolescentis - Corrected Proof

J. Wang, H. Zhao, W. Kong, C. Jin, Y. Zhao, Y. Qu, X. Xiao — 2009-12-07

Abstract: It was found that the intestinal bacteria balance would be deteriorated by rhubarb especially in long term treatment. Bifidobacteria is one of the most common species of probiotics in human intestine. The suppression of this particular probiotic, such as Bifidobacterium adolescentis, one of the dominant anaerobes in the intestines of humans, might lead to imbalance of intestinal flora and is considered to be potentially riskful for human health. Hence, the inhibitory effects of the five main components of hydroxyanthraquinones (HAQs) contained in rhubarb on B. adolescentis growth were investigated by microcalorimetry to discover the suppression potential of rhubarb and the structure-function relationship of such HAQs. The value of the maximum power- output (Pmax) and slope (k) of the thermogenic growth curves of B. adolescentis were found of decrease in the presence of the five HAQs, while the peak time (Tp) of the thermogenic curves were found to be delayed. The sequence of antimicrobial activity of the five HAQs is rhein>emodin>aloe-emodin>chrysophanol>physicion. The functional groups carboxyl, hydroxyl and hydroxylmethyl on phenyl ring in HAQs could improve the antimicrobial activity. The influence of substituent groups on anti- B. adolescentis activity might be related with the polarity and the sequence was carboxyl>hydroxyl>hydroxylmethyl>methyl and methoxyl.

Dammarenolic acid, a secodammarane triterpenoid from Aglaia sp. shows potent anti-retroviral activity in vitro - Corrected Proof

2009-12-07

Abstract: Screening of a panel of purified compounds isolated from Aglaia sp. (Meliaceae) for inhibition of early steps in the lentiviral replication cycle led to the identification of the 3, 4-secodammarane triterpenoid, ignT1, which inhibited HIV-1 infection potently (IC50=0.48μg/ml), while cytotoxic effects and inhibition of cell proliferation were only observed at concentrations exceeding 10.69μg/ml. Time of addition experiments revealed similar kinetics to the non-nucleoside RT-inhibitor (NNRTI), Nevirapine, although the latter was significantly less cytotoxic. However, unlike Nevirapine, dammarenolic acid also potently inhibited the in vitro replication of other retroviruses, including Simian immunodeficiency virus and Murine leukemic virus in vector-based antiviral screening studies. Interestingly, the methyl ester analogue of dammarenolic acid-methyldammarenolate had no anti-HIV-1 activity. Cell cycle analysis revealed that ignT1 arrests HeLa cells at the S and G2/M phase. These results strongly suggest that dammarenolic acid could be a promising lead compound for the development of novel anti-retrovirals.

Effects of inhaled Linalool in anxiety, social interaction and aggressive behavior in mice - Corrected Proof

2009-12-04

Abstract: Aromatherapy uses essential oils (EOs) for several medical purposes, including relaxation. The association between the use of aromas and a decrease in anxiety could be a valuable instrument in managing anxiety in an ever increasing anxiogenic daily life style. Linalool is a monoterpene commonly found as the major volatile component of EOs in several aromatic plant species. Adding to previously reported sedative effects of inhaled linalool, the aim of this study was to investigate the effects of inhaled linalool on anxiety, aggressiveness and social interaction in mice. Additionally, we investigated the effects of inhaled linalool on the acquisition phase of a step-down memory task in mice. Inhaled linalool showed anxiolytic properties in the light/dark test, increased social interaction and decreased aggressive behavior; impaired memory was only seen the higher dose of linalool. These results strengthen the suggestion that inhaling linalool rich essential oils can be useful as a mean to attain relaxation and counteract anxiety.

A dimeric high-molecular-weight chymotrypsin inhibitor with antitumor and HIV-1 reverse transcriptase inhibitory activities from seeds of Acacia confusa - Corrected Proof

S.K. Lam, T.B. Ng — 2009-12-04

Abstract: A dimeric 70-kDa chymotrypsin inhibitor with substantial N-terminal sequence homology to serine protease inhibitors was isolated from Acacia confusa seeds. The chymotrypsin inhibitor was purified using a protocol that entailed ion exchange chromatography on Q-Sepharose, SP-Sepharose and fast protein liquid chromatography-gel filtration on Superdex 75. The chymotrypsin inhibitor was unadsorbed on both Q-Sepharose and SP-Sepharose. Its chymotrypsin inhibitory activity was stable from pH 3 to 10 and from 0 to 50°C. It exerted antiproliferative activity toward breast cancer MCF-7 cells with an IC50 of 10.7±4.2μM. It inhibited HIV-1 reverse transcriptase with an IC50 of 8±1.5μM. It was devoid of antifungal activity toward a variety of fungal species. The distinctive features of the chymotrypsin inhibitor included dimeric nature, a high molecular mass, lack of trypsin inhibitory activity, highly potent HIV-1 reverse transcriptase inhibitory activity, specific antitumor activity and relatively high pH-stability.

Anti-diabetic effect of methylswertianin and bellidifolin from Swertia punicea Hemsl. and its potential mechanism - Corrected Proof

L.-Y. Tian, X. Bai, X.-H. Chen, J.-B. Fang, S.-H. Liu, J.-C. Chen — 2009-12-04

Abstract: In this study, we continued to investigate the hypoglycemic activity of Swertia punicea Helmsl., the hypoglycemic and hypolipidemic effects of methylswertianin and bellidifolin from the active ethyl acetate (EtOAc) fraction, and the potential mechanism(s) underlying the improvement of insulin resistance. Streptozotocin (STZ)-induced type 2 diabetic male BABL/c mice treated with methylswertianin and bellidifolin at different doses (orally, 200 and 100mg/kg body wt./day) for 4 weeks were analyzed in comparison to untreated mice. The results proved that methylswertianin and bellidifolin significantly reduced fasting blood glucose (FBG). The administration of both compounds also improved the oral glucose tolerance and lowered fasting serum insulin (FINS). Moreover, post-administration evaluation revealed lower serum total cholesterol (TC), low density lipoprotein cholesterol (LDL) and triglyceride (TG) levels and increased relative high density lipoprotein cholesterol (HDL) concentrations (HDL/TC). Methylswertianin and bellidifolin appeared to improve insulin resistance by enhancing insulin signaling. The expression levels of insulin-receptor α subunit (InsR-α), insulin-receptor substrate-1 (IRS-1), and phosphatidylinositol 3-kinase (PI3K) were also increased after administration. Meanwhile, methylswertianin and bellidifolin increased hepatic glycogen content, decreased glucokinase (GK) activities and increased glucose-6-phosphatase (G6Pase) activities. In conclusion, these result indicated that methylswertianin and bellidifolin could be useful for treating type-2 diabetes, likely via the improvement of insulin resistance (IR).

Wormwood (Artemisia absinthium) suppresses tumour necrosis factor alpha and accelerates healing in patients with Crohn’s disease – A controlled clinical trial - Corrected Proof

Simone Krebs, Talib N. Omer, Bilal Omer — 2009-12-04

Abstract: Suppression of tumour necrosis factor alpha (TNF-α) and other interleukins by wormwood (Artemisia absinthium) extracts were reported recently in in vitro studies. The aim of the present study was to find out if this effect can be also be observed in Crohn’s Disease (CD) patients where TNF-α appears to play an important role. In a controlled trial, 10 randomly selected patients suffering from CD were given in addition to their basic CD therapy 3×750mg dried powdered wormwood for 6 weeks. Ten patients, also randomly selected who met the inclusion criteria served as control group. Minimum score of 200 on Crohn’s Disease Activity Index (CDAI) was required at baseline for inclusion in each group. Patients who received infliximab or similar were excluded from the trial. TNF-α level in serum were measured at baseline, and after three and six weeks. During this period all concomitant CD medications was maintained at the baseline dose levels. Average serum TNF-α level fell from 24.5±3.5pg/ml at baseline to 8.0±2.5pg/ml after six weeks. The corresponding levels in the control group were 25.7±4.6 (week 0), and 21.1±3.2 (week 6). On the clinical side, CDAI scores fell from 275±15 to below 175±12 in wormwood group with remission of symptoms in eight patients (CDAI score below 170 or reduction by 70 points), compared to only two in the placebo group (CDAI of placebo group 282±11 at baseline and 230±14 on week 6). IBDQ also reflected accelerated clinical response with wormwood. Of clinical significance were the findings that wormwood also improved mood of the CD patients, as reflected in Hamilton's Depression Scale. These findings provide a base to test wormwood in clinical conditions thought to be mediated by increased production of pro-inflammatory cytokines such as TNF-α.

Effects of Hibiscus sabdariffa extract powder and preventive treatment (diet) on the lipid profiles of patients with metabolic syndrome (MeSy) - Corrected Proof

2009-12-04

Abstract: Insulin resistance, obesity, hypertension, and dyslipidemia are strongly associated with metabolic syndrome (MeSy), which is considered to be a reversible clinical stage before its evolution to coronary heart disease and diabetes. Currently, the antihypertensive and hypolipidemic properties of aqueous Hibiscus sabdariffa extracts (HSE) have been demonstrated in clinical trials and in vivo experiments. The aim of the present study was to evaluate the effects of a Hibiscus sabdariffa extract powder (HSEP) and a recognized preventive treatment (diet) on the lipid profiles of individuals with and without MeSy according to the National Cholesterol Education Program Adult Treatment Panel III (NCEP-ATP III) criteria. The protocol was a follow-up study carried out in a factorial, randomized design (T1=preventive treatment comprises Diet, T2=HSEP, T3=HSEP+preventive treatment (Diet) X MeSy, non-MeSy individuals). A total daily dose of 100 mg HSEP was orally administered in capsules for one month. The preventive treatment (diet) was selected according to NCEP-ATP III recommendations and adjusted individually. Total cholesterol, LDL-c, HDL-c, VLDL-c, triglycerides, glucose, urea, creatinine, AST, and ALT levels in the blood were determined in all individuals pre- and post-treatment. The MeSy patients treated with HSEP had significantly reduced glucose and total cholesterol levels, increased HDL-c levels, and an improved TAG/HDL-c ratio, a marker of insulin resistance (t-test p<0.05). Additionally, a triglyceride-lowering effect was observed in MeSy patients treated with HSEP plus diet, and in individuals without MeSy treated with HSEP. Significant differences in total cholesterol, HDL-c, and the TAG/HDL-c ratio were found when the means of absolute differences among treatments were compared (ANOVA p<0.02). Therefore, in addition to the well documented hypotensive effects of Hibiscus sabdariffa, we suggest the use of HSEP in individuals with dyslipidemia associated with MeSy.

Gedunin and Photogedunin of Xylocarpus granatum show significant anti-secretory effects and protect the gastric mucosa of peptic ulcer in rats - Corrected Proof

2009-12-04

Abstract: In the present study, the gastroprotective mechanism of Xylocarpus granatum fruit and its active constituents gedunin and photogedunin was investigated. Chloroform fraction (Fr-CHCl3) of X. granatum fruit was evaluated against cold restraint (CRU), aspirin (AS), alcohol (AL) and pyloric ligation (PL) induced gastric ulcer models in rats and histamine (HA) induced duodenal ulcer model in guinea pigs. Potential anti-ulcer activity of Fr-CHCl3 was observed against CRU (58.28%), AS (67.81%), AL (84.38%), PL (65.66%) and HA (61.93%) induced ulcer models. The standard drug omeprazole (10mg/kg, p.o.) showed 68.25% protection against CRU, 57.08% against AS and 69.42% against PL model and 70.79% against HA induced duodenal ulcer. Sucralfate, another standard drug (500mg/kg, p.o.) showed 62.72% protection in AL induced ulcer model. Fr-CHCl3 significantly reduced free acidity (51.42%), total acidity (30.76%) and upregulated mucin secretion by 58.37% respectively. Phytochemical investigations of Fr-CHCl3 yielded gedunin (36%), photogedunin (2%). Further, Fr-CHCl3 and its compounds gedunin and photogedunin significantly inhibited H+ K+-ATPase activity in vitro with IC50 of 89.37, 56.86 and 66.54μg/ml respectively as compared to the IC50 value of omeprazole (30.24μg/ml) confirming their anti-secretory activity. Conclusively, Fr-CHCl3 of Xylocarpus granatum was found to possess anti-ulcerogenic activity which might be due to its anti-secretory activity and subsequent strengthening of the defensive mechanism. This study is the first of its kind to show significant anti-secretory effect of gedunin and photogedunin. Therefore it could act as a potent therapeutic agent against peptic ulcer disease.

Cardioprotection with forsythoside B in rat myocardial ischemia-reperfusion injury: Relation to inflammation response - Corrected Proof

W.-L. Jiang, F.-H. Fu, B.-M. Xu, J.-W. Tian, H.-B. Zhu, Jian-Hou — 2009-12-03

Abstract: The present study was undertaken to examine the effect of forsythoside B (FB) on rat myocardial ischemia-reperfusion (I/R) model and elucidate the potential mechanism. Left ventricular systolic pressure (LVSP) and ±dp/dtmax were detected. Blood samples were collected to determine serum levels of troponin T (Tn-T), TNF-α and IL-6. Hearts were harvested to assess histopathological change and infarct size, determine content of MDA, myeloperoxidase (MPO), SOD and GPx activities, analyze expression of high-mobility group box 1 (HMGB1), phosphor-IκB–α and phosphor-nuclear factor kappaB (NF-κB) in ischemic myocardial tissue by Western blot. Compared with control group, rats treatment with FB showed a significant recovery in myocardial function with improvement of LVSP and ±dp/dtmax. The myocardial infarct volume, serum levels of Tn-T, TNF-α and IL-6, content of MDA and MPO activity in myocardial tissue were all reduced, protein expression of HMGB1, phosphor-IκB–α and phosphor-NF-κB were down-regulated, while attenuated the decrease of SOD and GPx activities. Besides, the infiltration of polymorph nuclear leukocytes (PMNs) and histopathological damages in myocardium were decreased in FB treated groups. These findings suggested that FB rescued cardiac function from I/R injury by limiting inflammation response and its antioxidant properties.

Antimicrobial activity and stability of rhinacanthins-rich Rhinacanthus nasutus extract - Corrected Proof

P. Puttarak, T. Charoonratana, P. Panichayupakaranant — 2009-11-02

Abstract: Rhinacanthins-rich Rhinacanthus nasutus (RRn) extract was prepared and standardized to contain total rhinacantins not less than 70% w/w. In this study, antimicrobial activities of the RRn extract as well as rhinacanthin-C against Streptococcus mutans, Propionibacterium acnes, Helicobacter pylori, Staphylococcus aureus, S. epidermidis and Candida albicans were evaluated by microdilution assay. It was found that the RRn extract exhibited potent bactericidal activity against S. mutans with MIC and MBC of 4μg/ml, and potent bacteriostatic activity against S. epidermidis, P. acnes and S. aureus with the MICs of 8–16μg/ml. However, the RRn extract was not active against C. albicans at concentration up to 2000μg/ml. The antimicrobial activities of the RRn extract was almost equal to those of rhinacanthin-C. Stability evaluations of the RRn extract through a period of 4 months found that the RRn extracts were stable when kept in a well-closed container protected from light and stored either under 4±2°C or 30±2°C. The RRn extracts that were exposed to light were not stable after 1 week of storage. Under accelerated conditions at 45°C with 75% relative humidity, the RRn extracts were not stable after 8 weeks of storage. Study on the effect of pH on the stability of the aqueous methanolic solution of the RRn extract found that the solutions were not stable at pH 5.5, pH 7.0, and pH 8.0. However, the rhinacanthin solution at pH 5.5 was more stable than those at pH 7.0 and pH 8.0.

Effects of Curcuma spp. on P-glycoprotein function - Corrected Proof

2009-11-02

Abstract: The effects of Curcuma longa (khamin chan) and Curcuma sp. “khamin-oi” (khamin-oi), as well as isolated major curcuminoids on intestinal P-gp functions were evaluated in vitro. The accumulation of R123 in Caco-2 cells was increased and the R123 efflux ratios were significantly decreased by both Curcuma longa and Curcuma sp. “khamin-oi” extracts, indicating their roles on efflux transporters. The a-b transport of daunorubicin was increased by curcumin, demethoxycurcumin and bisdemethoxycurcumin while the b-a transport was significantly decreased by curcumin and demethoxycurcumin. However, calcein-AM uptake into the human P-gp overexpression cell line, LLC-GA5-COL300, was increased by curcumin and demethoxycurcumin in a concentration-dependent manner but not affected by bisdemethoxycurcumin. These results show that curcumin and demethoxycurcumin could inhibit P-gp but bisdemethoxycurcumin may modulate the function of other efflux transporters such as MRP. Taken together, the information may indicate the impact of Curcuma longa and Curcuma sp. “khamin-oi” on pharmacokinetics of orally administered drugs that are P-gp substrates.

Antiproliferative, apoptotic and antimutagenic activity of isolated compounds from Polyalthia cerasoides seeds - Corrected Proof

Y.S. Ravikumar, K.M. Mahadevan, H. Manjunatha, N.D. Satyanarayana — 2009-10-30

Abstract: Phytochemical investigation of the petroleum ether extract fraction of Polyalthia cerasoides seeds led to the isolation of two phytosterols (α-spinasterol and spinasterol) and a clerodane di-terpenoid. The structures of these compounds were elucidated using IR, 1H-NMR, 13C-NMR and Mass spectral analysis. Further, these compounds were tested for antiproliferative action against CACO-2 cell line and apoptotic action was determined by nuclear staining and DNA fragmentation analysis. The results showed that the compounds exhibited antiproliferative action at various concentrations with an IC50 value of 28.6±4.34nM/ml, 57.7±6.81nM/ml and 60.0±7.10nM/ml for clerodane diterpenoid, spinasterol and α-Spinasterol respectively. Furthermore, the isolated compounds were screened for antimutagenic effect against methylmethane sulfonate (MMS) induced mutation. Phytosterols showed protective effect, whereas clerodane diterpenoid was less effective to MMS induced chromosomal aberrations. Our research contributes to the characterization of phytochemical constituents and to understand the ability of these compounds to antiproliferative and antimutagenic responses from the seed extracts.

Betacyanins from Portulaca oleracea L. ameliorate cognition deficits and attenuate oxidative damage induced by D-galactose in the brains of senescent mice - Corrected Proof

Chang-Quan Wang, Gui-Qin Yang — 2009-10-30

Abstract: This experiment was designed to assess the protective effect of betacyanins from Portulaca oleracea L. against the D-galactose (D-gal)-induced neurotoxicity in mice. Betacyanins from Portulaca oleracea markedly reversed the D-gal-induced learning and memory impairments, as measured by behavioral tests. The activities of superoxide dismutases (SOD), catalase (CAT), glutathione peroxidase (GPx) and glutathione reductase (GR) in D-gal-treated mice were enhanced, while the content of the lipid peroxidation product malondialdehyde (MDA) was decreased by betacyanin administration. Furthermore, significant negative correlations were found between mouse latency in finding the platform and the activities of SOD, CAT GR and GPx in the mouse brain, but the level of MDA correlated positively with the latency. These results suggest that the neuroprotective effect of betacyanins against D-gal-induced neurotoxicity might be caused, at least in part, by an increase in the activities of antioxidant enzymes with a reduction in lipid peroxidation. In comparison with vitamin C (VC), the betacyanins had a more pronounced effect on ameliorating cognition deficits in mice.

ACE inhibition by astilbin isolated from Erythroxylum gonocladum (Mart.) O.E. Schulz - Corrected Proof

2009-10-30

Abstract: Erythroxylum species have several traditional uses in different countries, including the treatment of hypertension. The ethanol extract from E. gonocladum aerial parts, a species endemic to the Brazilian cerrado, elicited a concentration-dependent inhibition of angiotensin converting enzyme (ACE) (pIC50=4.53±0.05). Extract fractionation led to the isolation of two compounds, whose structures were assigned by spectrometric data as astilbin and β-sitosterol, along with a mixture of palmitic, stearic and linolenic acids. This is the first report on the occurrence of these compounds on E. gonocladum. Astilbin promoted significant ACE inhibition in vitro (pIC50=5.86±0.33) and its activity did not differ from captopril, when both compounds were assayed at 10μM concentration.

Antileishmanial activity of furoquinolines and coumarins from Helietta apiculata - Corrected Proof

2009-10-30

Abstract: The bark infusion of H. apiculata are used to treat wound healing related to cutaneous leishmaniasis and as anti-inflammatory.Aim of the study: To isolate, purify active constituents of H. apiculata stem bark, and evaluate their in vitro and in vivo antileishmanial activities.Materials and methods: Isolation by chromatographic methods and chemical identification of furoquinoline alkaloids and coumarins, then evaluation of the in vitro leishmanicidal activity of these compounds against three strains of Leishmania sp. promastigotes and in vivo against Leishmania amazonensis in Balb/c mice.Results: Furoquinoline alkaloids and coumarins presented a moderate in vitro activity against promastigote forms of Leishmania sp. with IC50 values in the range between 17 and >50μg/ml. Balb/c mice infected with Leishmania amazonensis were treated with γ-fagarine by oral route, or with 3-(1’-dimethylallyl)-decursinol or (-)-heliettin by subscutaneous route for 14 days at 10mg/kg daily. In these conditions, γ-fagarine, 3-(1’-dimethylallyl)-decursinol and (-)-heliettin showed the same efficacy as the reference drug reducing by 97.4, 95.6 and 98.6% the parasite loads in the lesion, respectively.Conclusion: These compounds showed significant efficacy in L. amazonensis infected mice, providing important knowledge to improve its potential role for a future use in the treatment of cutaneous leishmaniasis.

Acanthoic acid, a diterpene in Acanthopanax koreanum, protects acetaminophen-induced hepatic toxicity in mice - Corrected Proof

2009-10-19

Abstract: The protective effect of a diterpenoid acanthoic acid (AA) isolated from Acanthopanax koreanum Nakai was investigated in acetaminophen (APAP)-induced hepatic toxicity. Drug-induced hepatotoxicity induced by an intraperitoneal (i.p.) injection of 300mg/kg (sub-lethal dose) of APAP. Pretreatment with AA (50 and 100mg/kg) orally 2h before the APAP administration attenuated the APAP-induced acute increase in serum aspartate aminotransferase (AST), and alanine aminotransferase (ALT) activites, replenished the depleted hepatic glutathione (GSH), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px) activities, decreased malondialdehyde (MDA) level and considerably reduced the histopathological alterations in a manner similar to silymarin (Sily). Immunohistochemical analyses also demonstrated that AA could reduce the appearance of necrosis regions as well as caspase-3 and hypoxia inducible factor-1α (HIF-1α) expression in liver tissue. Our results indicated that AA protected liver tissue from the oxidative stress elicites by APAP-induced liver damage and suggestes that the hepatic protection mechanism of AA would relate to antioxidation and hypoxia factor on APAP-induced hepatotoxicity.

Efficacy of components from leaves of Calophyllum brasiliense against Leishmania (Leishmania) amazonensis - Corrected Proof

2009-10-05

Abstract: Leishmanicide potential of Calophyllum brasiliense leaves on promastigote and amastigote of Leishmania (Leishmania) amazonensis is evaluated. The LD50 of dichloromethane extract and hexane fraction for promastigotes was respectively 40μg/ml and 20μg/ml. In mouse peritoneal macrophages infected with Leishmania amastigotes the Infection Index decreased respectively 100% and 84.2% in 80μg/ml and 40μg/ml concentrations of dichloromethane extract. Hexane fraction decreased infection index respectively by 98.7% and 91.3% within the same concentrations. It was found that pretreatment with dichloromethane extract or with hexane fraction of experimentally infected BALB/c mice decrease the volume of the lesions by L. (L.) amazonensis. Moreover, animals treated topically also revealed healing lesions. Besides, the parasite load in the animals’ popliteal lymph nodes was significantly reduced in treated animals, showing that plant components actually control infection. Results show that crude extract and hexane fraction of C. brasiliense reveal a significant in vitro and in vivo leishmanicide activity.

The alkaloid Berberine inhibits the growth of Anoikis-resistant MCF-7 and MDA-MB-231 breast cancer cell lines by inducing cell cycle arrest - Corrected Proof

2009-10-05

Abstract: Berberine is a pure phenanthren alkaloid isolated from the roots and bark of herbal plants such as Berberis, Hydrastis canadensis and Coptis chinensis. Berberine has been established to inhibit the growth of breast cancer cells, but its effects on the drug resistance and anoikis-resistance of breast cancer cells have yet to be elucidated. Anoikis, or detachment-induced apoptosis, may prevent cancer progression and metastasis by blocking signals necessary for survival of localized cancer cells. Resistance to anoikis is regarded as a prerequisite for metastasis; however, little is known about the role of berberine in anoikis-resistance. We established anoikis-resistant cells from the breast cancer cell lines MCF-7 and MDA-MB-231 by culturing them on a Poly-Hema substratum. We then investigated the effects of berberine on the growth of these cells. The anoikis-resistant cells had a reduced growth rate and were more invasive than their respective adherent cell lines. The effect of berberine on growth was compared to that of doxorubicine, which is a drug commonly used to treat breast cancer, in both the adherent and anoikis-resistant cell lines. Berberine promoted the growth inhibition of anoikis-resistant cells to a greater extent than doxorubicine treatment. Treatment with berberine-induced cell cycle arrest at G0/G1 in the anoikis-resistant MCF-7 and MDA-MB-231 cells as compared to untreated control cells. In summary, these results revealed that berberine can efficiently inhibit growth by inducing cell cycle arrest in anoikis-resistant MCF-7 and MDA-MB-231 cells. Further analysis of these phenotypes is essential for understanding the effect of berberine on anoikis-resistant breast cancer cells, which would be relevant for the therapeutic targeting of breast cancer metastasis.

Leishmanicidal activity of benzophenones and extracts from Garcinia brasiliensis Mart. fruits - Corrected Proof

2009-09-17

Abstract: Infections by protozoans of the genus Leishmania are the major worldwide health problem, with high endemicity in developing countries. The drugs of choice for the treatment of leishmaniasis are the pentavalent antimonials, which exert renal and cardiac toxicity. Thus, there is a strong need for safer and more effective treatments against leishmaniasis. The present study was designated to evaluate, by a bioguided assay, the leishmanicidal activity of extracts (hexane, ethyl-acetate and ethanolic) and molecules both obtained by means of extraction from pericarps of Garcinia brasiliensis fruits. The hexane extract presented the best activity on the extracellular (promastigotes) and intracellular (amastigotes) forms of Leishmania (L.) amazonensis, when compared to the other extracts. Based on these findings, this extract was fractionated by silica gel column chromatography, affording nine fractions then resulting in three purified prenylated benzophenones – 7-epi-clusianone (1), garciniaphenone (2) and guttiferone-a (3). They showed significant activity on Leishmania (L.) amazonensis, and little toxicity for mammalian cells. Structure-activity relationships were evaluated showing that the IC50 value displayed is dependent of prenyl groups and phenolic hydroxyls number, and inversely proportional to the hydrophobicity. Our results are promising, showing that these compounds are biologically active on Leishmania (L.) amazonensis.

Diallyl trisulfide induces Bcl-2 and caspase-3-dependent apoptosis via downregulation of Akt phosphorylation in human T24 bladder cancer cells - Corrected Proof

2009-09-14

Abstract: It is well known that the garlic-derived organosulfur compounds (OSCs) are effective to inhibit a variety of human cancers such as prostate, breast, colon, skin, lung, and bladder cancers. Herein, the pro-apoptotic effects of diallyl trisulfide (DATS), one of garlic-derived OSCs, on T24 bladder cancer cells were investigated. The results demonstrated that DATS suppressed proliferation of T24 bladder cancer cells in a dose- and time-dependent manner which was associated with induced G2/M Phase cell cycle arrest and apoptosis. Mechanistically, DATS inhibits phosphatidylinositol 3′-kinase/Akt activation that, in turn, results in modulation of Bcl-2 family proteins, leading to enhanced apoptosis of T24 cells. These findings suggest that DATS may be an effective way for treating human bladder and other types of cancers.

Anti-inflammatory activity of a HPLC-fingerprinted aqueous infusion of aerial part of Bidens tripartita L. - Corrected Proof

2009-09-14

Abstract: The anti-inflammatory potential of three doses of an aqueous infusion of aerial parts Bidens tripartita L. against carrageenan-induced acute paw edema in rats was investigated. A phytochemical study and qualitative-quantitative analyses revealed the presence of flavonoids, tannins, polysaccharides, phenols, amino acids, ascorbic acid, organic acids and polyacetylenes. Infusion doses of 20ml/kgbody wt. exhibited significant anti-inflammatory activity in rats, as compared with indomethacin. In addition, the infusion showed analgesic properties in a hot-plate test and antipyretic properties in carrageenan-induced local hyperthermia, both in rats. The effects were dose-dependent. Our results provide evidence for the potential usefulness of B. tripartita infusion in the treatment of inflammatory disorders.

Antiprotozoal activity of Betulinic acid derivatives - Corrected Proof

2009-09-14

Abstract: Betulinic acid (1), isolated from the crude extract of the leaves of Pentalinon andrieuxii (Apocynaceae), together with betulinic acid acetate (2), betulonic acid (3), betulinic acid methyl ester (4), and betulin (5) were evaluated for their antiprotozoal activity. The results showed that modifying the C-3 position increases leishmanicidal activity while modification of the C-3 and C-28 positions decreases trypanocidal activity.

WITHDRAWN: Hypoglycemic and hypolipidemic effect of a novel Gymnemic triacetate on STZ-induced diabetic rats - Corrected Proof

P. Daisy, J. Eliza, K.A. Mohamed Farook — 2009-09-14

This article has been withdrawn at the request of the author(s) and/or editor. The Publisher apologizes for any inconvenience this may cause.The full Elsevier Policy on Article Withdrawal can be found at http://www.elsevier.com/locate/withdrawalpolicy.

Xanthohumol enhances antiviral effect of interferon α-2b against bovine viral diarrhea virus, a surrogate of hepatitis C virus - Corrected Proof

Ni. Zhang, Zhengwen. Liu, Qunying. Han, Jinghong. Chen, Yi. Lv — 2009-09-14

Abstract: Xanthohumol (XN) is a natural compound with multifunctional potentials, including antiviral activity. In this study, the antiviral activity of addition of XN to interferon (IFN)-α was examined and compared with each compound alone using bovine viral diarrhea virus (BVDV), a surrogate model of hepatitis C virus (HCV). BVDV E2 protein and the viral RNA level were determined by immunofluorescence and quantitative real-time RT-PCR, respectively. The addition of XN to IFN-α significantly improved CPEs induced by the virus and inhibited BVDV E2 protein and viral RNA levels. The interaction between XN and IFN-α was significant (P<0.001). XN at 3.13μg/ml in combination with IFN-α at 50IU/ml showed greater inhibitory effect on the viral RNA level than each compound used alone at 6.25μg/ml and 100IU/ml, respectively, indicating synergistic effect on BVDV replication in this combination. The inhibitory activity in all the tested combinations of XN and IFN-α was stronger than that of each compound used alone at the corresponding concentration. These results suggest that XN in combination with IFN-α exhibited a greater in vitro antiviral effect compared with each compound used alone. Further studies are deserved to investigate the anti-HCV activity of XN and the potential of XN in formulating novel anti-HCV regimen.

Icariin isolated from Epimedium pubescens regulates osteoblasts anabolism through BMP-2, SMAD4, and Cbfa1 expression - Corrected Proof

Tsai-Pei Hsieh, Shiow-Yunn Sheu, Jui-Sheng Sun, Ming-Hong Chen, Man-Hai Liu — 2009-09-11

Abstract: Epimedii herba is one of the most frequently used herbs in formulas prescribed for the treatment of osteoporosis in China. The main active flavonoid glucoside extracted from Epimedium pubescens is Icariin, which has been reported to enhance bone healing and reduce osteoporosis occurrence. However, the detailed molecular mechanisms remain unclear. In this present study, we examine the molecular mechanisms of icariin by using primary osteoblast cell cultures obtained from adult mice.The osteoblast cells were harvested from 8-month old female Imprinting Control Region (ICR) mice. The effects of icariin stimulation on the proliferation, differentiation and maturation of osteoblasts were examined. The production of nitric oxide (NO) and caspase-3 were analyzed, along with the gene expressions of bone morphogenetic protein-2 (BMP-2), SMAD4, Cbfa1/Runx2, OPG, and RANKL.The viability of the osteoblasts reached its maximum at 10−8M icariin. At this concentration, icariin increased the proliferation and matrix mineralization of osteoblasts and promoted NO synthesis. With icariin treatment, the BMP-2, SMAD4, Cbfa1/Runx2, and OPG gene expressions were up-regulated; the RANKL gene expression was however down-regulated. Concurrent treatment involving the BMP antagonist (Noggin) or the NOS inhibitor (L-NAME) diminished the icariin-induced cell proliferation, ALP activity, NO production, as well as the BMP-2, SMAD4, Cbfa1/Runx2, OPG, RANKL gene expressions.In this study, we demonstrate that in vitro icariin is a bone anabolic agent that may exert its osteogenic effects through the induction of BMP-2 and NO synthesis, subsequently regulating Cbfa1/Runx2, OPG, and RANKL gene expressions. This effect may contribute to its action on the induction of osteoblasts proliferation and differentiation, resulting in bone formation.

Isolation and characterization of a French bean hemagglutinin with antitumor, antifungal, and anti-HIV-1 reverse transcriptase activities and an exceptionally high yield - Corrected Proof

S.K. Lam, T.B. Ng — 2009-09-09

Abstract: A dimeric 64-kDa hemagglutinin was isolated with a high yield from dried Phaseolus vulgaris cultivar “French bean number 35” seeds using a chromatographic protocol that involved Blue-Sepharose, Q-Sepharose, and Superdex 75. The yield was exceptionally high (1.1g hemagglutinin per 100g seed), which is around 10–85 times higher than other Phaseolus cultivars. Its N-terminal sequence resembled those of other Phaseolus hemagglutinins. The hemagglutinating activity of the hemagglutinin was stable in the pH range 6–8, and in the temperature range 0°C–50°C. It inhibited HIV-1 reverse transcriptase with an IC50 of 2μM. It suppressed mycelial growth in Valsa mali with an IC50 of 10μM. It inhibited proliferation of hepatoma HepG2 cells and breast cancer MCF-7 cells with an IC50 of 100 and 2μM, respectively. It had no antiproliferative effect on normal embryonic liver WRL68 cells.

Chemical constituents and antidepressant activity of the new species Hypericum enshiense occurring in China - Corrected Proof

Dongmei Wang, Jie Bai, Feng Sun, Depo Yang — 2009-08-25

Abstract: Hypericum enshiense L. H. Wu et F. S. Wang is a new species of Hypericum occurring in China, which was first identified and denominated by our laboratory. No research has been reported on the antidepressant activity and chemical constituents of this new species. In this study, the qualitative and quantitative analyses of the chemical constituents in the hydroalcoholic extract of this species were performed using HPLC/DAD/ESI-MS online method. Hypericin, pseudohypericin and some flavonoids were identified or tentatively identified. Furthermore, H. enshiense had a high content of hypericins than H. perforatum. In addition, the antidepressant activity of the hydroalcoholic extract of the species was investigated using forced swimming test (FST) and tail suspension test (TST). The extract significantly shortened the immobility time in FST and TST, while did not alter the locomoter activity of mice. These results suggested for the first time that the hydroalcoholic extract of H. enshiense might possess potential antidepressant-like activity in the animal behavioral models, and this species might act as a new potential resource for developing antidepressants to treat depressive disorders.