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One purified polysaccharide protein tyrosine phosphatase (PTP) was isolated from the roots of Polygala tenuifolia. The aim of the present study is to investigate the antitumor effect of PTP on human ovarian cancer OVCAR-3 cells and explore the molecular mechanism of the action involved. The results of MTT assay and apoptosis detection assay showed that PTP inhibited cellular proliferation of OVCAR-3 cells and induced apoptotic cellular death via arresting cell circle at the G0/G1 phase. Reverse transcription-polymerase chain reaction (RT-PCR) and Western blot analysis identified that bcl-2 gradually decreased at both transcription and protein levels after PTP treatment for 48 h in OVCAR-3 cells, while those of bax, cytochrome c, caspase-3, and caspase-9 increased. In addition, the low expression of NF-κB in PTP-treated OVCAR-3 cells would trigger the extrinsic pathway of programmed cell death signaling in tumor cells. These results together suggest that PTP may induce apoptosis of OVCAR-3 cells through a mitochondrial pathway.

In our previous study, we isolated a homogeneous polysaccharide (PTP) with antitumor activity from the roots of Polygala tenuifolia. In view of the close correlation between Bmi-1 expression and progression of ovarian cancer, we intend to elucidate the mechanism of its activity by determining the Bmi-1 expression and the telomerase activity in human ovarian carcinoma OVCAR-3 cells following treatment with PTP at three concentrations of 0.5, 1, and 2 mg/mL for 48 h. MTT and colony-forming assays revealed that PTP had a significant inhibitory effect on the cell growth and colony formation of OVCAR-3 cells. Furthermore, Western blot and real-time PCR analysis showed that PTP inhibited Bmi-1 both in protein and transcript levels. Besides, the telomerase activity in OVCAR-3 cells was also downregulated after PTP treatment for 48 h. Taken together, the inhibitory effect of PTP on the cell growth was at least in part mediated via the downregulation of Bmi-1 expression and the telomerase activity in OVCAR-3 cells, and PTP might be a new candidate for chemotherapeutic agent against human ovarian cancer.

The exploitation of various plant materials for the green synthesis of nanoparticles is considered an eco-friendly technology because it does not involve toxic chemicals. In this study, zinc oxide nanoparticles (ZnO NPs) were synthesized using the root extract of Polygala tenuifolia. Synthesized ZnO NPs were characterized by UV-Vis spectroscopy, FTIR, TGA, TEM, SEM and EDX. Anti-inflammatory activity was investigated in LPS-stimulated RAW 264.7 macrophages, whereas antioxidant activity was examined using a DPPH free radical assay. ZnO NPs demonstrated moderate antioxidant activity by scavenging 45.47% DPPH at 1mg/mL and revealed excellent anti-inflammatory activity by dose-dependently suppressing both mRNA and protein expressions of iNOS, COX-2, IL-1β, IL-6 and TNF-α.

Two new and five known acylated triterpene glycosides were isolated from the MeOH extract of the roots of Polygala tenuifolia. Based on extensive spectroscopic analysis, including 2D NMR experiments, and the results of alkaline hydrolysis, the structures of the new compounds were assigned as 3beta-[(beta-D-glucopyranosyl)oxy]-2 beta,27-dihydroxyolean- 12-ene-23,28-dioic acid 28-O-beta-D-apiofuranosyl-( 1--3)-[beta-D-galactopyranosyl-(1--4)-beta-D-xylopyranosyl-(1 -4)]-alpha-L-rhamnopyranosyl-(1-2)-3-O-(E)-3,4,5-trimeth oxycinnamoyl-beta-D-fucopyranosyl ester (1) and 3beta-[(beta-D-glucopyranosyl)oxy]-2beta,27-dihydroxyolean-1 2-ene-23,28-dioic acid 28-O-beta-D-apiofuranosyl-(l---3)-[beta-D-galactopyranosyl-(1--4)-beta-D-xylopyranosyl-( 1--4)]-alpha-L-rhamnopyranosyl-(1 -2)-[alpha-L-rhamnopyranosyl-( 1 -3 )]-4-O-(E)-3,4-dimethoxycinnamoyl-beta-D-fucopyranosyl ester (2).

isolation, structure determination and evaluation of inhibitory effects of five new phenolic glycosides, tenuisides A—E (I)—(III), new megastigmane glycoside, tenuiside F (IV), and seventeen known compounds on NO production in LPS-activated BV-2 microglia cells

The root ofPolygala tenuifoliaWilldenow has been used for the treatment of insomnia, depression, and amnesia. However, the toxicological properties of the herb have been overlooked, because it has been used for a long time for various purposes. In this study, we evaluated the preclinical safety of the root extract in rats and beagle dogs. First, the acute oral toxicity was tested in both rats and dogs. In the rats, only one female of 2 g/kg died, but no treatment-related death or clinical and gross findings were observed after the administration. No toxicological changes or mortalities related to the test substance were also observed after the administration in the dogs. Although vomiting, discoloration, or hemorrhage was found in some dogs, there were no serious abnormalities. Second, the subchronic toxicity was investigated in the rats. Two animals were found dead in the female group of 1,000 mg/kg/day, but there were no abnormal findings associated with the test substance. There also were no adverse effects on the clinical signs, body weight, and hematological and biochemical findings. Therefore, our results showed that the acute or subchronic toxicity of the root extract ofPolygala tenuifoliamight not be toxic to rats and dogs.

Learning and memory disorders arise from distinct age-associated processes, and aging animals are often used as a model of memory impairment. The root ofPolygala tenuifoliahas been commonly used in some Asian countries as memory enhancer and its memory improvement has been reported in various animal models. However, there is less research to verify its effect on memory functions in aged animals. Herein, the memory-enhancing effects of the crude extract ofPolygala tenuifolia(EPT) on normal aged mice were assessed by Morris water maze (MWM) and step-down passive avoidance tests. In MWM tests, the impaired spatial memory of the aged mice was partly reversed by EPT (100 and 200 mg/kg;P< 0.05) as compared with the aged control mice. In step-down tests, the nonspatial memory of the aged mice was improved by EPT (100 and 200 mg/kg;P< 0.05). Additionally, EPT could increase superoxide dismutase (SOD) and catalase (CAT) activities, inhibit monoamine oxidase (MAO) and acetyl cholinesterase (AChE) activities, and decrease the levels of malondialdehyde (MDA) in the brain tissue of the aged mice. The results showed that EPT improved memory functions of the aged mice probably via its antioxidant properties and via decreasing the activities of MAO and AChE.

Roots of Polygala tenuifolia Willdenow have been used in humans for centuries because of its sedative effects. We previously reported that BT-11, extracted from the roots of the plant, improved memory impairments in rats, enhanced memory in normal humans, and inhibited acetylcholinesterase activities in vitro. The present study was a randomized, double-blind, placebo-controlled comparison study to investigate whether BT-11 could enhance memory in the elderly humans. We used the Consortium to Establish a Registry for Alzheimer's Disease Assessment Packet (CERAD) and the Mini-Mental State Examination (MMSE). In the elderly, the total CERAD scores were much more significantly increased in the BT-11-treated group (n = 28) than in the placebo-treated group (n = 25). Especially, the mean scores of word list recognition, constructional recall and praxis, and modified Boston naming test were markedly improved in BT-11-treated group than in placebo-treated group. In conclusion, BT-11 could enhance some cognitive functions including memory in the elderly humans and therefore may be used as nutraceuticals that provide health benefits, including disease prevention and/or treatment.

We investigated whether an aqueous extract of Polygala tenuifolia root (PTAE) inhibits secretion of tumor necrosis factor-alpha (TNF-alpha) from primary cultures of mouse astrocytes. PTAE dose-dependently inhibited the TNF-alpha secretion by astrocytes stimulated with substance P (SP) and lipopolysaccharide (LPS). Interleukin-1 (IL-1) has been shown to elevate TNF-alpha secretion from LPS-stimulated astrocytes while having no effect on astrocytes in the absence of LPS. We therefore also investigated whether IL-1 mediated inhibition of TNF-alpha secretion from primary astrocytes by PTAE. Treatment of PTAE to astrocytes stimulated with both LPS and SP decreased IL-1 secretion to the level observed with LPS alone. Moreover, incubation of astrocytes with IL-1 antibody abolished the synergistic co-operative effect of LPS and SP. These results suggest that PTAE may inhibit TNF-alpha secretion by inhibiting IL-1 secretion and that PTAE has an anti-inflammatory activity on the central nervous system curing some pathological disease states.

Tenuifoliside A (TFSA) is a bioactive oligosaccharide ester component of Polygala tenuifolia Wild, a traditional Chinese medicine which was used to manage mental disorders effectively. The neuroprotective and anti-apoptotic effects of TFSA have been demonstrated in our previous studies. The present work was designed to study the molecular mechanism of TFSA on promoting the viability of rat glioma cells C6. We exposed C6 cells to TFSA (or combined with ERK, PI3K and TrkB inhibitors) to examine the effects of TFSA on the cell viability and the expression and phosphorylation of key proteins in the ERK and PI3K signaling pathway. TFSA increased levels of phospho-ERK and phospho-Akt, enhanced release of BDNF, which were blocked by ERK and PI3K inhibitors, respectively (U0126 and LY294002). Moreover, the TFSA caused the enhanced phosphorylation of cyclic AMP response element binding protein (CREB) at Ser133 site, the effect was revoked by U0126, LY294002 and K252a. Furthermore, when C6 cells were pretreated with K252a, a TrkB antagonist, known to significantly inhibit the activity of brain-derived neurotrophic factor (BDNF), blocked the levels of phospho-ERK, phospho-Akt and phosphor-CREB. Taking these results together, we suggested the neuroprotection of TFSA might be mediated through BDNF/TrkB-ERK/PI3K-CREB signaling pathway in C6 glioma cells.

Purpose: To gain insight into the mechanisms of analgesic and anti-inflammatory activities of the root extract of Polygala tenuifolia. Methods: Polygala tenuifolia was extracted with 70 % methanol and tested for analgesic and antiinflammatory activities (0.1, 1, 10 and 100 mg/kg) using the following models: acetic acid-induced writhing, rat paw edema, bradykinin inhibtion with rat ileum, and prostaglandin assay. Results: Administration of the Polygala tenuifolia extract at 100 mg/kg dose produced significant analgesic effect on acetic acid-induced writhing (97 % inhibition) but its effect in the tail-flick test was not significant (p < 0.05). In addition, the extract exerted significant anti-inflammatory effect in the rat paw edema model (8 to 33 % inhibition) at doses ranging from 0.1 - 100.0 mg/kg). A significant inhibitory action (53%) on the bradykinin-mediated contractions of rat ileum was also observed. Furthermore, the extract significantly (p < 0.05) inhibited the production of lipopolysaccharides-induced 6-keto-PGF1 by 28% in macrophage cultures. Conclusion: These results provide evidence that the Polygala tenuifolia root extract exerts analgesic and anti-inflammatory effects via its significant inhibitory effect on acetic acid writhing test, bradykinin-mediated actions as well as on 6-keto-PGF1 induction.

Objective To investigate the effect of Dingzhixiao Wan (DZXW), a classic traditional Chinese medicine formula consisting of Acorus tatarinowii, Polygala tenuifolia, Poria cocos and Panax ginseng in a proportion of 2: 2: 3: 3, on learning-memory impairment induced by scopolamine and its possible mechanisms. Method The mice were randomly divided into six groups: the control group, the model group, the positive huperzine A (0.05 mg x kg(-1)) group, DZXW 700 mg x kg(-1), 350 mg x kg(-1) and 175 mg kg(-1) groups. DZXW extracts were orally administrated to the mice for 7 days. Scopolamine (1.5 mg x kg(-1), ip) was injected to establish the learning and memory impairment model in mice. Morris water maze (MWM) test was used to assess the learning and memory ability of each group. After the test, the activities of glutamic acid (Glu), gamma-amino-butyric acid (GABA), serotonin (5-HT), dopamine (DA), acetylcholine (Ach) and acetyl cholinesterase (AchE) in brain tissue were measured. Result The praxiology test showed that DZXW significantly decreased the average latency of model mice in the place navigation test, and enhanced the frequency for passing through the platform in the spatial probe test, the percentage between target quadrant swimming distance and time. Moreover, DZXW could significantly increase the contents of Glu and 5-HT, DA and Ach, while reducing the levels of GABA and AchE in mice brain. Conclusion DZXW could significantly ameliorate the scopolamine-induced learning-memory impairment in mice and improve their learning-memory capacity, which may be related to its effect on adjusting Glu/GABA system and increasing Ach and monoamine neurotransmitter contents in mice brain.

Global warming has created opportunities and challenges for the survival and development of species. Determining how climate change may impact multiple ecosystem levels and lead to various species adaptations is necessary for both biodiversity conservation and sustainable biological resource utilization. In this study, we employed Maxent to predict changes in the habitat range and altitude of Polygala tenuifolia Willd. under current and future climate scenarios in China. Four representative concentration pathways (RCP2.6, RCP4.5, RCP6.0, and RCP8.5) were modeled for two time periods (2050 and 2070). The model inputs included 732 presence points and nine sets of environmental variables under the current conditions and the four RCPs in 2050 and 2070. The area under the receiver-operating characteristic (ROC) curve (AUC) was used to evaluate model performance. All of the AUCs were greater than 0.80, thereby placing these models in the "very good" category. Using a jackknife analysis, the precipitation in the warmest quarter, annual mean temperature, and altitude were found to be the top three variables that affect the range of P. tenuifolia. Additionally, we found that the predicted highly suitable habitat was in reasonable agreement with its actual distribution. Furthermore, the highly suitable habitat area was slowly reduced over time.

Two acidic polysaccharide fractions (PTPa and PTPb) extracted from the roots of Polygala tenuifolia, were obtained by DEAE-Sephacel anion-exchange, and Sephadex G-100 gel-permeation chromatography. High-performance liquid chromatography (HPLC) identified that PTPa and PTPb was composed of Ara, Glc, Gal, Man and GlcUA in the proportion of 2.4:1.2:0.6:0.4:1.1 and 2.1:1.7:0.5:0.6:1.7, respectively. Their molecular weight was evaluated to be 5.9 × 104 (PTPa) and 2.5 × 104 Da (PTPb) as determined by high performance size exclusion chromatography (HPSEC). Pharmacological studies revealed PTPa and PTPb significantly inhibited the growth of A549 cells in vitro and exhibited significantly higher antitumor activity against solid tumor A549 in vivo than did a blank control. Moreover, treatment with two acidic polysaccharides caused an enhancement of superoxide dismutase (SOD) and catalase (CAT) activities in tumor-bearing mice and a reduction in thiobarbituric acid reactive substances (TBARS) level. Taken together, these results indicated that two acidic polysaccharides from the roots of P. tenuifolia may be useful as potent antitumor agents for the prevention of lung tumorigenesis.

One polysaccharide PTP was isolated and purified from the roots of Polygala tenuifolia. It consisted of galactose, glucose and galactose in the ratio of 3.1:3.7:2.5, and a small amount of rhamnose, mannose and xylose. 17 general amino acids were identified to be components of the protein-bound polysaccharide analyzed by automatic amino acid analyzer. In order to test the anti-cancer activity of PTP, we investigated its effect against the growth of human ovarian cancer cells SKOV3 in vitro and in ovarian cancer rats. The intracellular reactive oxygen species (ROS) and glutathione (GSH) in SKOV3 cells following PTP treatment were also quantified to explore the possible mechanism underlying the antitumor activity of the polysaccharide. The result showed that PTP is effective on inhibiting the proliferation of SKOV3 cells in a concentration-dependent manner. Furthermore, treatment with PTP caused a rapid depletion of intracellular GSH content and accumulation of intracellular ROS, thus resulting in the apoptosis, which may prove to be a pivotal mechanism for its cancer protection action. In addition, a significant tumor growth inhibition effect was observed in nude mice after PTP administration for 7 weeks. All above indicated PTP could be beneficial towards ovarian cancer therapy.

Дана эколого-фитоценотическая характеристика мест обитания P. tenuifolia. Изучены морфогенез особей и онтогенетическая структура ценопопуляции этого вида в различных эколого-фитоценотических условиях. Установлено, что в ходе развития у особей последовательно сменяются две фазы морфогенеза: первичный побег, первичный куст. Полученные данные по онтогенезу особей и некоторым особенностям биологии вида позволяют смоделировать характерный онтогенетический спектр. Результаты по изучению онтогенетической структуры ценопопуляций с применением комплексного подхода дают возможность оценить современное состояние ценопопуляций в Республиках Хакасия и Тыва. Установлен тип популяционной стратегии P. tenuifolia.

BACKGROUND: The root of Polygala tenuifolia, a traditional Chinese medicine, has been used to improve memory and intelligence, while the underlying mechanisms remain largely unknown. In this study, we investigated the protective effects of senegenin, an component of Polygala tenuifolia root extracts, on cognitive dysfunction induced by hepatic ischemia-reperfusion. METHODOLOGY/PRINCIPAL FINDINGS: Initially, we constructed a rat model of hepatic ischemia-reperfusion (HIR) and found that the memory retention ability of rats in the step-down and Y maze test was impaired after HIR, paralleled by a decrease of N-methyl-D-aspartate (NMDA) receptor NR2B subunit mRNA and protein expressions in hippocampus. Furthermore, we found that administration of senegenin by gavage attenuated HIR-induced cognitive impairment in a dose and time dependent manner, and its mechanisms might partly due to the increasing expression of NR2B in rat hippocampus. CONCLUSIONS/SIGNIFICANCE: Cognitive dysfunction induced by HIR is associated with reduction of NR2B expression. Senegenin plays a neuroprotective role in HIR via increasing NR2B expression in rat hippocampus. These findings suggest that senegenin might be a potential agent for prevention and treatment of postoperative cognitive dysfunction (POCD) or other neurodegenerative diseases.

The present study was to investigate the influence of tenuigenin, an active ingredient of Polygala tenuifolia Willd, on the proliferation and differentiation of hippocampal neural stem cells in vitro. Tenuigenin was added to a neurosphere culture and neurosphere growth was measured using MTT assay. The influence of tenuigenin on the proliferation of neural progenitors was examined by Clone forming assay and BrdU detection. In addition, the differentiation of neural stem cells was compared using immunocytochemistry for β III-tubulin and GFAP. The results showed that addition of tenuigenin to the neural stem cell medium increased the number of newly formed neurospheres. More neurons were also obtained when tenuigenin was added in the differentiation medium. These findings suggest that tenuigenin is involved in regulating the proliferation and differentiation of hippocampal neural stem cells. This result may be one of the underlying reasons for tenuigenin's nootropic and anti-aging effects.

Pentobarbital was used to induce the place orientation and space exploration dysfunctions. Morris water maze was used to observe mice's abilities of place orientation and space exploration. Bradford method was used for the determination of the total protein and spectrometry method was used for the detection of acetylcholinesterase (AChE) activity. Extract of Polygala tenuifolia and Poria cocos (EPP) could shorten the escape latency in place orientation test and increase numbers of mice's crossing the location where the platform had been in the space exploration test. EPP could decrease AChE activities in the brain and the serum of mice with learning and memory disorders induced by pentobarbital. EPP would improve the learning and memory abilities of mice induced by chemical drugs. The results demonstrate that EPP would improve place orientation and space exploration dysfunctions of mice and it; exerts its effects perhaps by inhibiting AChE activity to reduce acetylcholine (ACh) hydrolysis in the brain and the serum in mice

In order to find natural agents against Dactylogyrus intermedius in goldfish, petroleum ether, chloroform, ethyl acetate, acetone, and methanol extracts of three medicinal plants (Dryopteris crassirhizoma, Kochia scoparia, and Polygala tenuifolia) were screened for antiparasitic properties using in vivo anthelmintic efficacy assay. Among these extracts investigated, methanolic extract of D. crassirhizoma was observed the most effective with EC50 value of 22.97 mg L−1 after 48 h of exposure, which exhibited a 100% efficacy against D. intermedius at 60.00 mg L−1, followed by the methanolic extracts of K. scoparia and P. tenuifolia with EC50 values of 31.28 and 154.79 mg L−1, showing 100% efficacy against D. intermedius at 60.00 and 500.00 mg L−1, respectively. In addition, acute toxicity assay indicated that 48-h LC50 values of methanolic extracts of D. crassirhizoma, K. scoparia, and P. tenuifolia were 4.10-, 2.27-, and 5.00-fold higher than the corresponding EC50. The obtained results demonstrated that methanolic extracts of D. crassirhizoma, K. scoparia, and P. tenuifolia have the potential for the development of novel therapy for the control of D. intermedius in aquaculture.

A methanolic extract of the roots of Polygala tenuifolia (Polygalaceae) significantly attenuated nitric oxide (NO) production in lipopolysaccharide (LPS)-stimulated BV2 microglia cells. Five xanthones, 1-hydroxy-7-methoxyxanthone (1), 3,6-dihydroxy-1,2,7-trimethoxyxanthone (2), 1,3,6-trihydroxy-2,7-dimethoxyxanthone (3), 1,7-dihydroxy-2,3-dimethoxyxanthone (4) and 1,7-dihydroxy-3-methoxyxanthone (5), and five phenylpropanoids, 4-hydroxy-3-methoxypropiophenone (6), methyl 4-hydroxy-3-methoxycinnamic acid (7), 3,4,5-trimethoxycinnamic acid (8), 4-methoxycinnamic acid (9) and β-d-(3-O-sinapoyl) fructofuranosyl-α-d-(6-O-sinapoyl)glucopyranoside (10), were isolated from CHCl(3) fraction using bioactivity-guided fractionation. Among these compounds, compounds 1, 2, 4, 5 and 7 showed significant inhibitory effects on LPS-induced NO production in BV2 microglia cells at the concentration ranging from 10.0 to 100.0 μM.

Three phenylpropenoyl sucroses – sibiricose A5 (1), A6 (2) and 3′,6-disinapoyl sucrose (3) – were isolated from the 30% EtOH extract of Polygala tenuifolia, which displayed antidepressant-like action. HPLC analysis indicated that the three phenylpropenoyl sucroses could be absorbed into serum. From the serum pharmacochemistry point of view, these three phenylpropenoyl sucroses might prevent or relieve depression.

a b s t r a c t Aim of the study: The root of Polygala tenuifolia Willd is a well-known traditional Oriental medicine and Q2 has been prescribed for treatment of dysfunction in memorial systems and various brain inflammatory diseases. The present study was designed to validate the anti-inflammatory effects of the water extract of Polygala tenuifolia root (WEPT). Materials and methods: The anti-inflammatory properties of WEPT were studied using lipopolysaccha- ride (LPS)-stimulated murine BV2 microglia model. As inflammatory parameters, the production of nitric oxide (NO), inducible NO synthase (iNOS), cyclooxygenase (COX)-2, prostaglandin E2 (PGE2), tumor necro- sis factor (TNF)-, and interleukin (IL)-1 were evaluated. We also examined the extract's effect on the activity of nuclear factor-kappaB (NF-B), and toll-like receptor 4 (TLR4) and myeloid differentiation factor 88 (Myd-88) expression. Results: WEPT suppressed LPS-induced production of NO, PGE2, and expression of iNOS and COX-2 in a dose-dependent manner, without causing cytotoxicity. It also significantly reduced generation of proin- flammatory cytokines, including IL-1 and TNF-. In addition, WEPT suppressed NF-B translocation by blockade of IkappaB- (IB-) degradation and inhibited TLR4 and Myd-88 expression in LPS-stimulated BV2 cells. Conclusions: These results indicate that the inhibitory effects of WEPT on LPS-stimulated inflammatory mediator production in BV2 microglia are associated with suppression of the NF-B and toll-like receptor signaling pathways. Therefore, Polygala tenuifolia extracts may be useful in treatment of neurodegener- ative diseases by inhibition of inflammatory mediator production in activated microglia. © 2011 Published by Elsevier Ireland Ltd.

A new xanthone C-glycoside, polygalaxanthone III (1), and a new acylated sugar, tenuifoliside E (2) were isolated from the roots of Polygala tenuifolia. Their structures were characterized as 4-C-[beta-D-apiofuranosyl-(1-->6)-beta-D-glucopyranosyl]-1, 3,6-trihydroxy-7-methoxyxanthone (1) and beta-D-(1-O-acetyl-3-O-feruloyl-6-O-sinapoyl)-fructofuranosy l-alpha-D-(2,4,6- O-triacetyl)glucopyranoside (2), respectively, on the basis of chemical and spectral evidence including two dimensional nuclear magnetic resonance (2D-NMR) studies.

The constituents of the ethanol extract from the root of Polygala tenuifolia Willd. (Polygalaceae) were investigated for antioxidant activity in senescence-accelerated mice. Consequently, two relevant samples were obtained, a fraction separated by macroporous resin (YZ-OE), and a major pure crystal of 3,6'-disinapoyl sucrose (DISS). Based on HPLC-ESI-MS analysis, the most constituents in the YZ-OE fraction from the extract of P. tenuifolia were oligosaccharide esters. The antioxidant activities of these two samples were evaluated using the accelerated senescence-prone, short-lived mice (SAMP) in vivo. The activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-PX) were increased significantly in SAMP mice fed oligosaccharide esters (YZ-OE 50 mg/kg) and its constituents (DISS 50 mg/kg). However, the content of malondialdehyde (MDA) was increased in the blood and liver of SAMP mice. But when given YZ-OE, it could be decreased, by 44.3% and 47.5%, respectively, compared with the SAMP model. Results from the analyses indicated that the oligosaccharide esters (YZ-OE) from roots of P. tenuifolia had a high in vivo antioxidant activity.

YZ-50 is an active fraction obtained from the root of Polygala tenuifolia Willd. (Polygalaceae) extract and it has been reported previously to exert beneficial effects on mental health in depressed sufferers, however, its mechanism of action remains unresolved. This study utilized the chronic mild stress (CMS) model of depression in Sprague-Dawley rats to evaluate the effects of YZ-50 on depressive behaviors. Furthermore, we tested the hypothesis that the capacity of YZ-50 to reverse the harmful effects of CMS is relative to the hypothalamo-pituitary-adrenal (HPA) system and brain-derived neurotrophic factor (BDNF) in the hippocampus. Repeated administration of YZ-50 for 28 days at the doses of 140 and 280 mg/kg in CMS, YZ-50 reversed the CMS-induced changes in sucrose consumption, plasma corticosterone levels and open field activity. In addition, CMS significantly decreased hippocampal BDNF mRNA levels. However, YZ-50 counteracted a decrease in hippocampal BDNF mRNA caused by CMS. In conclusion, YZ-50 reversed the harmful effects of CMS on mood and behaviors in rats and it possesses an antidepressant property that is at least in part mediated by the neuroendocrine and neuropropective systems, and it is likely that the HPA system plays an important role in this process.

Aim of study Modified Yeoldahanso-tang (MYH) is a Korean herbal formula, containing 10 herbs: Pueraria lobata (Willd.) Ohwi, Angelica tenuissima Nakai, Scutellaria baicalensis Georgi, Platycodon grandiflorum (Jacq), Angelicae Dahurica, Cimicifuga heracleifolia Kom, Raphanus sativa L., Polygala tenuifolia (Willd.), Acorus gramineus Soland. and Dimocarpus longan Lour. The constitutive ratio of the ten herbs is at 6:4:2:1:2:2:2:4:6:6 in dry weight. MYH has been used to treat amnesia, hypochondria and dementia in Korea. In this study, we explored the possibility of using MYH in the prevention and treatment of Parkinson's disease (PD). Specifically, we made an effort to demonstrate the neuroprotective effects of MYH using experimental methods similar to those used in a recent study of PD. Materials and methods 1-Methyl-4-phenylpyridinium (MPP+) (400 μM) was used to induce cytotoxicity in NGF (nerve growth factor)-differentiated PC12 cells. Cell viability was measured using a MTT assay. Induction of autophagy by MYH in NGF-differentiated PC12 cells was measured using an immunoblotting assay with LC3 and beclin 1 antibodies. The proteasomal inhibitor lactacystin (10 μM) was used to cause UPS dysfunction in NGF-differentiated PC12 cells. Clearance of aggregated proteins by MYH was measured using an immunoblotting assay with an ubiquitin antibody. 1-Methyl-4-phenyl-1,2,3,6-tetrahydrophenylpyridine (MPTP) (20 mg/kg, 4 times i.p.) caused substantia nigra injuries in C57BL/6 mice. Dopamine (DA) neurons were identified using a tyrosine hydroxylase-immunohistochemistry (TH-IHC) assay with a rabbit anti-TH antibody. Results Our findings indicate that MYH provides protection against MPP+-induced injury in NGF-differentiated PC12 cell. And MYH provides neuroprotection against lactacystin-induced NGF-differentiated PC12 cell death, which effect is partially mediated by autophagy enhancement through enhanced degradation of aggregated proteins. Additionally, in a C57BL/6 mice model with MPTP-induced substantia nigra injuries, MYH inhibits both the loss of TH-positive neurons in the substantia nigra pars compacta (SNpc) and the reduction of the optical density of TH-IR fibers in the striatum (ST). Conclusions All of our results indicate that MYH treatment has neuroprotective effects that are partially mediated by autophagy enhancement. MYH may be a promising herbal formula for the prevention and treatment of neurodegenerative diseases, especially PD.

Aim of the study Chong–Myung–Tang (CMT) consisted of Acorus gramineus Soland, Polygala tenuifolia Willdenow, and Poria cocos Wolf is one of the traditional Korean herbal medicines used for the therapy of learning and memory improvement. The present study was investigated the effect of CMT on learning and memory functions in SCOP-induced memory deficits mice. Materials and methods The cognitive-enhancing effect of CMT on amnesic mice induced by SCOP was investigated by assessing the passive avoidance test and the Morris water maze test. In order to confirm the underlying mechanisms of memory enhancing effects of CMT, activities of AChE, choline acetyltransferase (ChAT), and antioxidant enzymes were measured. Results Administration of CMT significantly restored memory impairments induced by SCOP in the passive avoidance test and also reduced escape latency during trial sessions in the Morris water maze test. The increased AChE activity produced by SCOP was significantly inhibited by CMT. CMT significantly enhanced ChAT activity. Moreover, treatment with CMT to the amnesic mice induced by SCOP considerably decreased malondialdehyde levels and restored activities of superoxide dismutase and catalase to the control values. Conclusions These results suggest that CMT may be useful for the cognitive improvement via regulation of cholinergic marker enzyme activities and the antioxidant defense system.

From the roots of Polygala tenuifolia WILLD. ten new oligosaccharides, called tenuifolioses G-P, were isolated and their structures were elucidated by spectroscopic data and chemical evidance. These oligosaccharides were esterified with acetic, benzoic, p-coumaric and/or ferulic acid.

From the roots of Polygala tenuifolia , six new xanthones, 3-hydroxy-2,8-dimethoxyxanthone, 7-hydroxy-1,2,3-trimethoxyxanthone, 3-hydroxy-1,2,7-trimethoxyxanthone, 1,7-dimethoxy-2,3-methylenedioxyxanthone, 6,8-dihydroxy-1,2,4-trimethoxyxanthone, and 6,8-dihydroxy-1,2,3-trimethoxyxanthone, have been isolated along with seven known xanthones, 1,7-dihydroxyxanthone, 1,7-dimethoxyxanthone, 1-hydroxy-3,7-dimethoxyxanthone, 2,3,8-trimethoxyxanthone, 1,7-dihydroxy-2,3-dimethoxyxanthoner, 1,6-dihydroxy-3,7-dimethoxyxanthone and 1,3,6,7-tetramethoxyxanthone. Their structures have been elucidated by chemical and spectroscopic evidence.

The aim of this study was to investigate the cognition-enhancing activity and underlying mechanisms of a triterpenoid saponin (polygalasaponin XXXII, PGS32) isolated from the roots of Polygala tenuifolia Willd. The Morris water maze was used to evaluate the spatial learning and memory of mice. To detect the basic properties of synaptic transmission and long-term potentiation (LTP) in the dentate gyrus of rats, electrophysiological recordings were made of evoked potentials. Western blotting analysis and immunofluorescence assays were used to determine the phosphorylation of extracellular signal-regulated kinase (ERK), cAMP response element-binding protein (CREB), synapsin I and the expression of brain derived neurotrophic factor (BDNF). When administered at 0.125, 0.5, or 2 mg/kg, PGS32 could significantly prevent scopolamine-induced cognitive impairments in mice. Intracerebroventricular (icv) administration of PGS32 greatly enhanced basic synaptic transmission in the dentate gyrus of rats and induced LTP. In primary hippocampal neurons, as well as in the hippocampus of maze-trained mice, PGS32 activated the mitogen-activated protein (MAP) kinase cascade by promoting phosphorylation of ERK, CREB and synapsin I. The expression of BDNF was also greatly enhanced in the hippocampus. Our findings suggest that PGS32 can improve hippocampus-dependent learning and memory, possibly through improvement of synaptic transmission, activation of the MAP kinase cascade and enhancement of the level of BDNF. Therefore, PGS32 shows promise as a potential cognition-enhancing therapeutic drug.

Ethnopharmacological relevance SH21B is an anti-obesity composition composed of seven herbs: Scutellaria baicalensis Georgi, Prunus armeniaca Maxim, Ephedra sinica Stapf, Acorus gramineus Soland, Typha orientalis Presl, Polygala tenuifolia Willd and Nelumbo nucifera Gaertner. It has been used for the treatment of obesity in traditional medical clinics in Korea, but its anti-obesity effects and mechanism of action have not been studied until now. Aim of the study This study was conducted to confirm the anti-obesity effects of SH21B and to elucidate its mechanism of action. Materials and methods 3T3-L1 adipocytes and high fat diet-induced obese mice were treated with SH21B, and its effect on gene expression was analyzed using microarray technology, real-time PCR and western blotting experiments. Results SH21B significantly inhibited fat accumulation in 3T3-L1 adipocytes and reduced adipose tissue and serum triglyceride levels in high fat diet-induced obese mice. Microarray analyses showed that SH21B affected more genes in the adipogenesis pathway than any other pathway studied. SH21B significantly decreased the expression of major transcription factors of the adipogenesis pathway and resulted in the down-regulation of lipid metabolizing enzymes involved in the transport, uptake and synthesis of lipids. Conclusions SH21B inhibits fat accumulation by down-regulating the expression of genes involved in the adipogenesis pathway.

Radix Polygalae (“Yuan Zhi”, the roots of Polygala tenuifolia Willd., YZ) is an important herb used in traditional Chinese medicine to mediate depression. The present study was designed to verify the antidepressant effects of the standardized YZ ethanol extract (YZE) and its four fractions YZ-30, YZ-50, YZ-70 and YZ-90 on the tail suspension (TST) and forced swimming test (FST). Furthermore, the standardization of the fractions obtained from the separation procedures was carried out by high-performance liquid chromatography (HPLC)-fingerprint. The YZ-50 fraction (Oligosaccharide esters –enriched, oral (200 mg/kg) showed a significant anti-immobility like effects. The data of YZ-50 on the corticosterone-induced injure of SH-SY5Y human neuroblastoma cell indicated that YZ-50 may have biological effects on neuroprotection. Proliferation of cell lines was assessed by dimethylthiazoldiphenyltetrazoliumbromide (MTT) and 5-bromo-2′-deoxyuridine (BrdU) incorporation assays. It was found that YZ-50 and its two bioactive compounds, 3,6′-di-o-sinapoyl-sucrose (DISS) and tenuifoliside A(TEA) showed protection activities in SY5Y cells from the lesion. By using bioassay-screening methods, our results indicate that the presence of oligosaccharide esters such as DISS and TEA in this herb may be responsible for the cytoprotective activity effects.

Aim Previous studies have shown that tenuigenin, a crude extract of Polygala tenuifolia Willd. that is commonly used in traditional Chinese herbal medicine for memory loss, can reduce the secretion of Abeta from cultured cells. However, the mechanism underlying this effect and the active compound derived from tenuigenin is unknown. In this study, a purified component of tenuigenin, tenuifolin, was examined and revealed to be an effective compound in vitro. Methods Abeta secretion from three sets of COS-7 cells, each carrying a plasmid expressing a different form of APP was examined following the treatment with tenuifolin. Initially, tenuifolin was determined to have no inherent toxicity to either the transfected or wild type cells at the effective concentrations. Cells were then treated with 0.5-2.0 microg mL(-1) tenuifolin for 12 h and their media were examined via an ELISA for Abeta1-40 and Abeta-42. Results We found that treatment with 2.0 microg mL(-1) tenuifolin significantly decreased Abeta secretion from COS-7 cells without altering the ratio of Abeta1-40 and Abeta-42. This effect is most probably due to inhibition of the beta-site APP cleaving enzyme as Abeta secretion was not inhibited from cells expressing the C99 fragment. Conclusion Tenuifolin is an effective compound from tenuigenin. We believe that this finding should lead the way for future experiments to determine the exact mechanism for tenuifolin's effect on Abeta secretion.

Four new phenolic glycosides, tenuifolisides A (1), B (2), C (3), and D (4) together with a known phenolic glyciside, β-D-(3-O-sinapoyl)-fructofuranosyl-α-D-(6-O-sinapoyl)-glucopyranoside (5) were isolated from the roots of Polygala tenuifolia. The structures of these new compounds were characterized as β-D-(3-O-(3, 4, 5-trimethoxycinnamoyl)]-fructofuranosyl-α-D-[6-O-(p-hydroxybenzoyl)]-glucopyranoside (1), β-D-(3-O-sinapoyl)-fructofuranosyl-α-D-(6-O-[p-hydroxybenzoyl)]-glucopyranoside (2), β-D-[3-O-(3, 4, 5-trimethoxycinnamoryl)]-fructofuranosyl-α-D-(6-O-sinapoyl)-glucopyranoside (3), and 1, 5-anhydro-[6-O(3, 4, 5-trimethoxycinnamoryl)]-D-glucitol (4), respectively, on the basis of chemical and spectral evidence including two dimensional unclear magnetic resonance (2D-NMR) studies.

The methanol fraction of an ethanolic extract from the roots of Polygala tenuifolia Willd. showed antagonistic action on neurotoxicity induced by glutamate and serum deficiency in PC12 cells. Bioassay-guided fractionation led to the isolation of six new triterpenoid saponins, onjisaponins V - Z, and Vg ( 1 - 6), together with ten known saponins ( 7 - 16). The structures of 1 - 6 were elucidated by spectroscopic and chemical methods. Screening results indicated that compounds 1 - 16 showed neuroprotective effects against serum deficiency and glutamate at the concentration of 10 (-5) mol/L.

Eighteen different triterpene saponins isolated from Polygala tenuifolia were investigated by electrospray ionization ion trap multiple-stage mass spectrometry (ESI-ITMS(n)) in positive and negative ion modes. MS(1)-MS(3)/MS(4) spectra of the both modes were analyzed, and they all gave fragments in line and shared common fragmentation patterns. Key fragments from MS(n) spectra of both the modes and their proposed fragmentation pathways were constructed with examples illustrated for the formation of characteristic fragments in the saponins. Two special fragmentation patterns were proposed: (1) the formation of fragments by cleavage of CH(2)O from Delta(12)-14alpha-CH(2)OH of the oleanene-type saponin aglycone in both positive and negative MS(n) (n > or = 2) modes; (2) the occurrence of fragments by cleavage of CO(2) and 3-glucose as the characteristic structure feature of 23-COOH at the oleanene-type saponin aglycones coupled with 3-Glc substitutes in the negative MS(n) (n > or = 2) modes. Peak intensities in MS(n) spectra were also correlated with structural features and fragmentation preferences of the investigated saponins, which are discussed in detail. In general, fragments formed predominantly by cleavages of glycosidic bonds in the positive mode, while selective cleavages of acyl bonds preceded that of glycosidic bonds in negative MS(n) (n > or = 2) mode, both of which could well be applied to the structural analysis of these saponins. Interpretation of MS(n) spectra presented here provided diagnostic key fragment ions important for the structural elucidation of saponins in P.tenuifolia.

We previously demonstrated that tenuifoliside B and 3,6'-disinapoylsucrose in Polygalae Radix, the root of Polygala tenuifolia WILLDENOW, inhibited potassium cyanide (KCN)-induced hypoxia and scopolamine-induced memory impairment in mice. Because both ingredients have a common sinapoyl moiety in their structure, we inferred that the sinapoyl moiety could inhibit hypoxia and memory impairment. In the present study to clarify the hypothesis, sinapic acid inhibited KCN-induced hypoxia and scopolamine-induced memory impairment as well as tenuifoliside B and 3,6'-disinapoylsucrose did. In addition, sinapic acid inhibited decompression- or bilateral carotid artery ligation-induced hypoxia (or mortality) and CO 2 -induced impairment in mice, and basal forebrain lesion-induced cerebral cholinergic dysfunction (decreases in acetylcholine concentration and choline acetyltransferase activity) in rats. These results, taken together, suggest the possibilities that sinapic acid is not only a very important moiety in the pharmacological activities of tenuifoliside B and 3,6'-disinapoylsucrose but also a candidate for a cerebral protective and cognition-improving medicine.

Although Polygala tenuifolia WILLD (PT) was classically mentioned as an anti-dementia drug in Chinese and Japanese traditional medicine, basic research showed only enhancement of the cholinergic function. In Alzheimer's disease, neuritic atrophy and synaptic loss occur prior to neuronal death event, and may be the first trigger of the memory impairment. Therefore, we studied effects of Polygala tenuifolia WILLD (PT) on Abeta(25-35)-induced neuronal damage using rat cortical neurons for characterization of activities of PT under Abeta-induced neuronal damage. Treatment with the water extract of PT enhanced axonal length dose-dependently after Abeta(25-35)-induced axonal atrophy. However, dendritic atrophy and synaptic loss induced by Abeta(25-35) were not recovered by treatment with PT extract. In contrast, Abeta(25-35)-induced cell damage was completely inhibited by PT extract. By characterization of PT effects on neuronal morphological plasticity and cell damage, usefulness as well as an insufficiency of PT as an anti-dementia drug was clarified.

Two new acylated presenegenin glycosides E-onjisaponin H (5) and Z-onjisaponin (6) together with seven known saponins were isolated from the roots of Polygala tenuifolia Willd. Compounds 5 and 6 were obtained as a pair of isomers due to trans and cis-p-methoxycinnamoyl. Their structures were elucidated mainly by 2D-NMR techniques including 1H-1HCOSY, TOCSY, HSQC, HMBC as 3-O-(beta-D-glucopyranosyl) presenegenin 28-[O-beta-D-apiofuranosyl-(1 --> 3)-O-[beta-D-xylopyranosyl-(1 --> 4)]-O-alpha-L-rhamnopyranosyl-(1 --> 2)-O-[alpha-L-rhamnopyranosyl-(1 --> 3)]-4-O-[(E)-p-methoxycinnamoyl]-beta-D-fucopyranosyl] ester (5) and its (Z)-isomer (6).

Polygalae Radix (PR) from Polygala tenuifolia (Polygalaceae) is traditionally used in China and Korea, as this herb has a sedative, anti-inflammatory and antibacterial agent. To extend our understanding of the pharmacological actions of PR in the CNS on the basis of its CNS inhibitory effect, the present study examined whether PR has the neuroprotective action against N-methyl-D-aspartate (NMDA)-induced cell death in primarily cultured rat cerebellar granule neurons. PR, over a concentration range of 0.05 to 5 μg/ml, inhibited NMDA (1 mM)-induced neuronal cell death, which was measured by a trypan blue exclusion test and a 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide (MTT) assay. PR (0.5 μg/ml) inhibited glutamate release into medium induced by NMDA (1 mM), which was measured by HPLC. Pre-treatment of PR (0.5 μg/ml) inhibited NMDA (1 mM)-induced elevation of intracellular Ca2+concentration ([ Ca2+]i), which was measured by a fluorescent dye, Fura 2-AM, and generation of reactive oxygen species (ROS). These results suggest that PR prevents NMDA-induced neuronal cell damage in vitro.

We studied the cognitive improving and cerebral protective constituents in the roots of Polygala tenuifolia WILLDENOW, a well-known Chinese traditional medicine prescribed for amnesia, neurasthenia, palpitation, noctural emission and insomnia. Tenuifoliside B (1), which is one of the acylated oligosaccharides in the roots of P. tenuifolia, showed the cerebral protective effect on potassium cyanide (KCN)-induced anoxia in mice, widely used as an animal model for cerebrovascular disease, and also had an ameliorative effect on the scopolamine-induced impairment of performance in passive avoidance task in rats. Compound 1 significantly enhanced oxotremorine-induced tremors in mice, suggesting that it ameliorated the scopolamine-induced impairment of passive avoidance response by enhancing the cholinergic system. These findings show that compound 1 has cognitive improving and cerebral protective effects.

The effect of PAP 9704, a traditional prescription in Korea consisting of Polygala tenuifolia, Acorus gramineus, and Poria cocos at a ratio of 1:1:1 (dry weight), on methamphetamine (MA)-induced hyperlocomotion was examined in mice. The increased locomotor activity induced by MA (1 mg/kg/d, i.p. x 7) was significantly attenuated by co-administration with PAP 9704 (100 or 200 mg/kg/d, p.o. x 7) in a dose dependent manner. Consistently, it was found that the hyperlocomotor activity occurred in parallel with the expression of striatal fos-related antigen immunoreactivity. The adenosine A(2A) receptor antagonist, 1,3,7-trimethyl-8-(3-chlorostyryl)xanthine (0.5 or 1.0 mg/kg, i.p.), significantly reversed the pharmacological action of PAP 9704 in a dose related manner, but the adenosine A(1) receptor antagonist 8-cyclopentyl-1,3-dimethylxanthine (0.5 or 1.0 mg/kg, i.p.) and the A(2B) receptor antagonist alloxazine (1.5 or 3.0 mg/kg, i.p.) did not significantly affect this pharmacological action. Our results suggest that PAP 9704 prevents MA-induced hyperlocomotion, at least in part, via the stimulation of the adenosine A(2A) receptor.