Polyphyllin VI Research Articles

Assessment of the inhibition risk of paris saponins, bioactive compounds from Paris polyphylla, on CYP and UGT enzymes via cocktail inhibition assays

Paris saponins, also known as polyphyllins, are natural compounds extracted from Paris polyphylla, which have many pharmacological activities, such as anti-inflammation and anti-cancer. In particular, paris saponin I, II, VII and polyphyllin VI are the components of the quality standard for Paris polyphylla. However, the inhibition risk of polyphyllins on cytochrome P450 (CYP) and UDP-glucuronosyltransferases (UGT) remains unclear. Therefore, this report investigated the potential inhibitory effects of paris saponin I, II, VII and polyphyllin VI on the activities of CYP (CYP1A2, CYP2B1, CYP2C11, CYP2D1, CYP2E1 and CYP3A2) and UGT (UGT1A1, UGT1A3, UGT1A6, PROG and AZTG) through cocktail inhibition assays in vitro. In the study of CYP, polyphyllin VI exhibited weak inhibition on CYP2D1 activity in rat liver microsomes with IC50 value at 45.2 μM, while paris saponin VII weakly inhibited CYP2C11 and CYP2E1 activities with IC50 value at 42.0 and 67.7 μM, respectively. In the study of UGT, none of the four steroidal saponins showed significant inhibition risk. In conclusion, paris saponin I, II, VII and polyphyllin VI have very low potential to cause the possible toxicity and drug interactions involving CYP and UGT enzymes, indicating that they are safe enough to take with drugs.

Polyphyllin VI Induces Apoptosis and Autophagy via Reactive Oxygen Species Mediated JNK and P38 Activation in Glioma.

BackgroundPolyphyllin VI (PPVI), a bioactive component derived from a traditional Chinese herb Paris polyphylla, exhibits potential antitumor activity against hepatocellular carcinoma, as well as breast and lung cancers. However, its effect on glioma remains unknown.MethodsFive glioma cell lines (U251, U343, LN229, U87 and HEB) and an animal model were employed in the study. Anti-proliferation effects of PPVI were first determined using CCK-8 cell proliferation and clone formation assays, then reactive oxygen species (ROS), cell cycle progression and apoptosis effects measured by flow cytometry. The effect of PPVI on protein expression was quantified by Western blot analysis.ResultsData showed that PPVI inhibited the proliferation of glioma cell lines by modulating the G2/M phase. Additionally, incubation of cells with PPVI promoted apoptosis, autophagy, increased accumulation of ROS and activated ROS-modulated JNK and p38 pathways. On the other hand, N-acetyl cysteine, a ROS inhibitor, attenuated PPVI-triggered effects. Furthermore, JNK and p38 inhibitors ameliorated PPVI-triggered autophagy and apoptosis in glioma cells. In vivo assays showed that PPVI inhibited tumor growth of U87 cell line in nude mice.ConclusionOverall, these data suggested that PPVI might be an effective therapeutic agent for glioma.

Anthelmintic efficacy of 35 herbal medicines against a monogenean parasite, Gyrodactylus kobayashii, infecting goldfish (Carassius auratus)

Species of Gyrodactylus are common monogenean ectoparasites of fish and can result in severe economic losses in aquaculture. To find effective and environmentally acceptable therapies to control gyrodactylosis in aquaculture, anthelmintic efficacy of 35 kinds of herbal medicines was investigated against Gyrodactylus kobayashii infecting goldfish (Carassius auratus), and the promising herbal medicines were further isolated by bioassay-guided fractionation. Among the tested extracts, the methanol extract of Paris polyphylla was the most effective and showed 100% anthelmintic activity against G. kobayashii at 20 mg/L, and had a therapeutic index (TI) of 37.9, which is higher than that of widely used formaldehyde-based parasiticides in aquaculture. According to high-performance liquid chromatography (HPLC) analysis, six active compounds were identified: dioscin, polyphyllin I, gracillin, polyphyllin II, polyphyllin VII and polyphyllin VI; among these active compounds, dioscin was the most effective in eliminating Gyrodactylus infections, with EC50 value of 0.17 mg/L and TI value of 8.18, and exhibiting 100% anthelmintic efficacy at 0.5 mg/L. In order of efficacy, polyphyllin I and gracillin had EC50 values of 0.21 and 0.19 mg/L and TI values of 5.35 and 4.97, respectively. These findings suggested that dioscin, polyphyllin I and gracillin are effective and safe in the treatment of G. kobayashii infections and might become leading natural compounds for the synthesis and development of therapeutic agents against monogenean parasites in aquaculture. Interestingly, the methanol extract of P. polyphylla had a higher TI value of 37.9 compared to the abovementioned active compounds, although it required higher concentrations to reach 100% anthelmintic efficacy. The results indicated the methanol extract of P. polyphylla was less toxic to fish than the three compounds mentioned above, and although its anthelmintic activity was inferior to that of these compounds, the methanol extract was a relatively safe and beneficial agent for practical application to treat gyrodactylosis in aquaculture.

Polyphyllin VI Induces Caspase-1-Mediated Pyroptosis via the Induction of ROS/NF-κB/NLRP3/GSDMD Signal Axis in Non-Small Cell Lung Cancer.

Trillium tschonoskii Maxim (TTM), a traditional Chinese medicine, has been demonstrated to have a potent anti-tumor effect. Recently, polyphyllin VI (PPVI), a main saponin isolated from TTM, was reported by us to significantly suppress the proliferation of non-small cell lung cancer (NSCLC) via the induction of apoptosis and autophagy in vitro and in vivo. In this study, we further found that the NLRP3 inflammasome was activated in PPVI administrated A549-bearing athymic nude mice. As is known to us, pyroptosis is an inflammatory form of caspase-1-dependent programmed cell death that plays an important role in cancer. By using A549 and H1299 cells, the in vitro effect and action mechanism by which PPVI induces activation of the NLRP3 inflammasome in NSCLC were investigated. The anti-proliferative effect of PPVI in A549 and H1299 cells was firstly measured and validated by MTT assay. The activation of the NLRP3 inflammasome was detected by using Hoechst33324/PI staining, flow cytometry analysis and real-time live cell imaging methods. We found that PPVI significantly increased the percentage of cells with PI signal in A549 and H1299, and the dynamic change in cell morphology and the process of cell death of A549 cells indicated that PPVI induced an apoptosis-to-pyroptosis switch, and, ultimately, lytic cell death. In addition, belnacasan (VX-765), an inhibitor of caspase-1, could remarkably decrease the pyroptotic cell death of PPVI-treated A549 and H1299 cells. Moreover, by detecting the expression of NLRP3, ASC, caspase-1, IL-1β, IL-18 and GSDMD in A549 and h1299 cells using Western blotting, immunofluorescence imaging and flow cytometric analysis, measuring the caspase-1 activity using colorimetric assay, and quantifying the cytokines level of IL-1β and IL-18 using ELISA, the NLRP3 inflammasome was found to be activated in a dose manner, while VX-765 and necrosulfonamide (NSA), an inhibitor of GSDMD, could inhibit PPVI-induced activation of the NLRP3 inflammasome. Furthermore, the mechanism study found that PPVI could activate the NF-κB signaling pathway via increasing reactive oxygen species (ROS) levels in A549 and H1299 cells, and N-acetyl-L-cysteine (NAC), a scavenger of ROS, remarkably inhibited the cell death, and the activation of NF-κB and the NLRP3 inflammasome in PPVI-treated A549 and H1299 cells. Taken together, these data suggested that PPVI-induced, caspase-1-mediated pyroptosis via the induction of the ROS/NF-κB/NLRP3/GSDMD signal axis in NSCLC, which further clarified the mechanism of PPVI in the inhibition of NSCLC, and thereby provided a possibility for PPVI to serve as a novel therapeutic agent for NSCLC in the future.

Polyphyllin VI, a saponin from Trillium tschonoskii Maxim. induces apoptotic and autophagic cell death via the ROS triggered mTOR signaling pathway in non-small cell lung cancer

Non-small cell lung cancer (NSCLC) accounts for approximately 85% of all lung cancers. Our previous studies have proven that Trillium tschonoskii Maxim. (TTM), a traditional Chinese medicine, possesses potent anti-tumor effect. However, the detailed components and molecular mechanism of TTM in anti-NSCLC are still unknown. In the present experiment, polyphyllin VI (PPVI) was successfully isolated from TTM with guidance of the anti-proliferative effect in A549 cells, and the cell death of PPVI treated A549 and H1299 cells was closely linked with the increased intracellular ROS levels. In addition, PPVI induced apoptosis by promoting the protein expression of Bax/Bcl2, caspase-3 and caspase-9, and activated autophagy by improving LC3 II conversion and GFP-LC3 puncta formation in A549 and H1299 cells. The mechanism study found that the activity of mTOR which regulates cell growth, proliferation and autophagy was significantly suppressed by PPVI. Accordingly, the PI3K/AKT and MEK/ERK pathways positively regulating mTOR were inhibited, and AMPK negatively regulating mTOR was activated. In addition, the downstream of mTOR, ULK1 at Ser 757 which downregulates autophagy was inhibited by PPVI. The apoptotic cell death induced by PPVI was confirmed, and it was significantly suppressed by the overexpression of AKT, ERK and mTOR, and the induced autophagic cell death which was depended on the Atg7 was decreased by the inhibitors, such as LY294002 (LY), Bafilomycin A1 (Baf), Compound C (CC) and SBI-0206965 (SBI). Furthermore, the mTOR signaling pathway was regulated by the increased ROS as the initial signal in A549 and H1299 cells. Finally, the anti-tumor growth activity of PPVI in vivo was validated in A549 bearing athymic nude mice. Taken together, our data have firstly demonstrated that PPVI is the main component in TTM that exerts the anti-proliferative effect by inducing apoptotic and autophagic cell death in NSCLC via the ROS-triggered mTOR signaling pathway, and PPVI may be a promising candidate for the treatment of NSCLC in future.

<p>Polyphyllin VI induces apoptosis and autophagy in human osteosarcoma cells by modulation of ROS/JNK activation</p>

PurposePolyphyllin VI, a main active saponin isolated from traditional medicinal plant Paris polyphylla, has exhibited antitumor activities in several cancer cell lines. In the present study, we investigated the antitumor effect of Polyphyllin VI against human osteosarcoma cells (U2OS) and the underlying molecular mechanisms.MethodsThe U2OS cell lines were used to determine the antiproliferative effect of Polyphyllin VI by CCK8 assay. Cell cycle was analyzed by flow cytometry. The Polyphyllin VI-induced apoptosis was determined by Annexin V-APC/7-AAD apoptosis detection kit and JC-1 staining. Meanwhile, the autophagy was determined by acridine orange staining. The apoptosis and autophagy-related proteins were monitored by Western blot assay. Subsequently, intracellular hydrogen peroxide (H2O2) and the activation of ROS/JNK pathway were detected.ResultsPolyphyllin VI could potently inhibit cell proliferation by causing G2/M phase arrest. Polyphyllin VI induced mitochondria-mediated apoptosis with the upregulation of proapoptotic proteins Bax and poly ADP-ribose polymerase, and downregulation of antiapoptotic protein Bcl-2 in U2OS cells. Concomitantly, Polyphyllin VI provoked autophagy with the upregulation of critical Atg proteins and accumulation of LC3B-II. Intracellular H2O2 production was triggered upon exposure to Polyphyllin VI, which could be blocked by ROS scavenger. Polyphyllin VI dramatically promoted JNK phosphorylation, whereas it decreased the levels of phospho-p38 and ERK.ConclusionOur results reveal that Polyphyllin VI may effectively induce apoptosis and autophagy to suppress cell growth via ROS/JNK activation in U2OS cells, suggesting that Polyphyllin VI is a potential drug candidate for the treatment of osteosarcomas.

Paris Polyphylla Inhibits Colorectal Cancer Cells via Inducing Autophagy and Enhancing the Efficacy of Chemotherapeutic Drug Doxorubicin.

Colorectal cancer is one of the most common cancers worldwide and chemotherapy is the main approach for the treatment of advanced and recurrent cases. Developing an effective complementary therapy could help to improve tumor suppression efficiency and control adverse effects from chemotherapy. Paris polyphylla is a folk medicine for treating various forms of cancer, but its effect on colorectal cancer is largely unexplored. The aim of the present study is to investigate the tumor suppression efficacy and the mechanism of action of the ethanolic extract from P. polyphylla (EEPP) in DLD-1 human colorectal carcinoma cells and to evaluate its combined effect with chemotherapeutic drug doxorubicin. The data indicated that EEPP induced DLD-1 cell death via the upregulation of the autophagy markers, without triggering p53- and caspase-3-dependent apoptosis. Moreover, EEPP treatment in combination with doxorubicin enhanced cytotoxicity in these tumor cells. Pennogenin 3-O-beta-chacotrioside and polyphyllin VI were isolated from EEPP and identified as the main candidate active components. Our results suggest that EEPP deserves further evaluation for development as complementary chemotherapy for colorectal cancer.

Targeted regulation of Rell2 by microRNA-18a is implicated in the anti-metastatic effect of polyphyllin VI in breast cancer cells

Polyphyllin VI (PP-VI) is one of the major saponins present in Paris polyphylla Sm., a medicinal plant primarily used for cancer treatment in China and India. However, its anti-metastatic activity remains largely unknown. The current study thus investigated the anti-metastatic activity of PP-VI in mouse mammary carcinoma 4T1 and human breast cancer MDA-MB-231 cells. The anti-metastatic effect of PP-VI was investigated at a sub-cytotoxic dose in migration and invasion assays in vitro. Experimental metastasis mouse model was used to examine the anti-metastatic effect of PP-VI in vivo. Additionally, target prediction, real-time PCR, Western blotting and luciferase assay were performed to identify the target gene of a pro-metastatic microRNA, miR-18a in 4T1 cells. The effect of PP-VI on the identified target of miR-18a was further investigated. The results showed that PP-VI impaired the viability of 4T1 and MDA-MB-231 cells. Moreover, when applied at a sub-cytotoxic dose, PP-VI suppressed the metastatic potential of 4T1 and MDA-MB-231 cells. Receptor expressed in lymphoid tissue (RELT)-like 2 (Rell2) was identified as a direct target of miR-18a with anti-metastatic functions in 4T1 and MDA-MB-231 cells. PP-VI treatment resulted in increased expression of Rell2 and decreased level of miR-18a in 4T1 and MDA-MB-231 cells. PP-VI treatment also attenuated miR-18a mimic or Rell2 siRNA-augmented migration of MDA-MB-231 cells. The current work thus demonstrates for the first time that targeted regulation of Rell2 by miR-18a is in part implicated in the anti-metastatic effect of PP-VI in breast cancer cells, providing novel pharmacological insights into the anti-cancer effect of PP-VI.

Molecular Mechanisms of Apoptosis in HepaRG Cell Line Induced by Polyphyllin VI via the Fas Death Pathway and Mitochondrial-Dependent Pathway.

Polyphyllin VI, which is an active saponin, is mainly isolated from traditional medicinal plant Paris polyphylla, which causes liver damage in rats. In the present study, we aimed to explore the potential cytotoxicity of polyphyllin VI on the growth of HepaRG cells and to determine the molecular mechanism. The results revealed that polyphyllin VI changed cell morphology and induced apoptosis in HepaRG cells. Flow cytometric assay displayed that polyphyllin VI promoted the generation of reactive oxygen species (ROS), depolarized the mitochondrial membrane potential (MMP), and induced S phase cell cycle arrest by decreasing the expression of cyclin A2 and CDK2, while significantly increasing the expression of p21 protein. Polyphyllin VI induced the release of cytochrome c from the mitochondria to the cytosol and activated Fas, caspase-3, -8, -9, and PARP proteins. Pretreatment with NAC and Z-VAD-FMK (ROS scavenger and caspase inhibitor, respectively) on HepaRG cells increased the percentage of viable cells, which indicated that polyphyllin VI induced cell apoptosis through mitochondrial pathway by the generation of ROS and Fas death-dependent pathway. All of the effects are in dose- and time-dependent manners. Taken together, these findings emphasize the necessity of risk assessment to polyphyllin VI and offer an insight into polyphyllin VI-induced apoptosis of HepaRG cells.

Quantitative evaluation and discrimination of wild Paris polyphylla var. yunnanensis (Franch.) Hand.-Mazz from three regions of Yunnan Province using UHPLC-UV-MS and UV spectroscopy couple with partial least squares discriminant analysis.

Paris polyphylla var. yunnanensis (Franch.) Hand.-Mazz (PPY) is used widely as an anthelmintic, antimicrobial, and anti-tumor agent. Multiplicate analytical methods have been employed to discriminate PPY from different regions, as well as to identify regions most beneficial to the growing of this species. In this study, a convenient and accurate method was established using ultra high performance liquid chromatography (UHPLC) for simultaneous determination of four steroid saponins (Pa, Pb, polyphyllin VI, and chonglou saponin VII). Partial least squares discriminant analysis (PLS-DA) according to UHPLC and UV spectroscopy was applied to analyze 30 samples of PPY from three regions of Yunnan Province in China, and identify significant peaks. The results indicated that the correlation coefficients (r 2) of all calibration curves were above 0.999, and the inter- and intra-day relative standard deviations (RSD) of retention time and peak areas of common peaks were below 1.78% and 3.40%, respectively, with recovery rates of 99.6-103.4% with RSD≤2%. Quantitative analysis implied that the average values of total saponins in PPY from south Yunnan Province (19.9mg/g) were higher than in the central (8.79mg/g) district. Thus, further investigation could focus on the southern region to seek high quality PPY. The analysis found that PLS-DA for ultraviolet (UV) spectroscopy, which could separate the samples from three regions, was more appropriate than UHPLC. Retention times during 20-30.75min of UHPLC, and absorption at 200-300nm of the UV spectrum were identified as significant peaks for distinguishing PPY from different regions.

Anti-lung Cancer Effects of Polyphyllin VI and VII Potentially Correlate with Apoptosis In Vitro and In Vivo.

Polyphyllin VI (PVI) and polyphyllin VII (PVII) derived from Paris polyphylla possess anti-cancer activities. However, the mechanisms for the anti-lung cancer effects of PVI and PVII remain poorly understood. In this study, PVI and PVII exhibited inhibitory effects on the proliferation of A549 and NCI-H1299 cells. PVI and PVII induced G2/M cell cycle arrest and triggered apoptosis. PVI and PVII upregulated the tumor suppressor protein p53 and downregulated cyclin B1. The two treatments significantly increased the expression levels of death receptor 3, death receptor 5, Fas, cleaved PARP, and cleaved caspase-3. Furthermore, PVI and PVII significantly inhibited the growth of A549 cells in vivo. The tumor inhibitory rates of PVI were 25.74%, 34.62%, and 40.43% at 2, 3, and 4 mg/kg, respectively, and those of PVII were 25.63%, 41.71%, and 40.41% at 1, 2, and 3 mg/kg, respectively. Finally, PVI and PVII regulated the expression of proteins related to the apoptotic pathway in A549 xenografts. In summary, PVI and PVII exhibited strong inhibitory effects on lung cancer cell growth in vitro and in vivo by inducing G2/M cell cycle arrest and triggering apoptosis.

Discrimination of wild Paris based on near infrared spectroscopy and high performance liquid chromatography combined with multivariate analysis.

Different geographical origins and species of Paris obtained from southwestern China were discriminated by near infrared (NIR) spectroscopy and high performance liquid chromatography (HPLC) combined with multivariate analysis. The NIR parameter settings were scanning (64 times), resolution (4 cm−1), scanning range (10000 cm−1∼4000 cm−1) and parallel collection (3 times). NIR spectrum was optimized by TQ 8.6 software, and the ranges 7455∼6852 cm−1 and 5973∼4007 cm−1 were selected according to the spectrum standard deviation. The contents of polyphyllin I, polyphyllin II, polyphyllin VI, and polyphyllin VII and total steroid saponins were detected by HPLC. The contents of chemical components data matrix and spectrum data matrix were integrated and analyzed by partial least squares discriminant analysis (PLS-DA). From the PLS-DA model of NIR spectrum, Paris samples were separated into three groups according to the different geographical origins. The R2X and Q2Y described accumulative contribution rates were 99.50% and 94.03% of the total variance, respectively. The PLS-DA model according to 12 species of Paris described 99.62% of the variation in X and predicted 95.23% in Y. The results of the contents of chemical components described differences among collections quantitatively. A multivariate statistical model of PLS-DA showed geographical origins of Paris had a much greater influence on Paris compared with species. NIR and HPLC combined with multivariate analysis could discriminate different geographical origins and different species. The quality of Paris showed regional dependence.

Polyphyllin VI inhibits colitis associated colorectal carcinogenesis in mice: Possible mechanisms

Polyphyllin VI inhibits colitis associated colorectal carcinogenesis in mice: Possible mechanisms

Effect of recombinant human endostatin on VEGF and TIMP-1 expression in hypetrophic scar of rabbit ears

Objective To introduce an accelerated solvent extraction(ASE)-HPLC-TOF/MS method for simultaneous determination of Polyphyllin Ⅰ,Polyphyllin Ⅱ,Polyphyllin Ⅵ,Polyphyllin Ⅶ,Dioscin and Gracillin in Paris Polyphylla.Methods The extraction was performed by ASE under the following conditions: extracting solvent 70% ethanol;extracting temperature 120℃;pressure 1.17 MPa(1 700 psi);and static time 6 min.The separation was carried out on a MGC18 column(3.0 mm×100 mm,3.0 μm).Elution was done with a linear gradient mobile phase system consisting of 0.1% formic acid(V/V) water and acetonitrile.The flow rate was set at 0.8 ml/min,the sample injection volume was 3 μl,the column temperature was kept constant at 25℃,nebulizer gas pressure was 275.8 kPa(40 psi),drying gas flow rate was 10 L/min,and gas temperature was 350℃.Results The calibration curves for Polyphyllin Ⅰ,Polyphyllin Ⅱ,Polyphyllin Ⅵ,Polyphyllin Ⅶ,Dioscin and Gracillin were linear within the range of 0.500 0-50.00 μg/ml(r=0.999 8),0.422 3-42.23 μg/ml(r=0.999 7),0.612 0-61.20 μg/ml(r=0.999 9),0.714 0-71.40 μg/ml(r=0.999 5),0.448 0-44.80 μg/ml(r=0.999 9) and 0.436 0-43.60 μg/ml(r=0.9997),respectively;and the average recoveries(n=6) were 98.9%,98.0%,102.5%,101.9%,103.1% and 97.9%,respectively.Conclusion The introduced method is solvent-saving and time-saving;it can also be used for high-throughput analysis and determination of steroidal saponins in Paris Polyphylla.

Liquid chromatography tandem mass spectrometry in study of the pharmacokinetics of six steroidal saponins in rats

A rapid, simple and sensitive high-performance liquid chromatography tandem mass spectrometry method was developed and validated for simultaneous determination of six main steroidal saponins in Paris polyphylla in rat plasma. Ginsenoside Rg3 was selected as the internal standard (IS). Plasma samples were pretreated with protein precipitation, and the separation was achieved on a reverse phase Agilent poroshell120 EC-C18 column using a gradient mobile phase system of acetonitrile–water containing 0.1% formic acid. The triple quadruple mass spectrometer was set in negative electrospray ionization mode and multiple reaction monitoring (MRM) was used for six steroidal saponins quantification. The precursors to produce ion transitions monitored for polyphyllin I, polyphyllin II, polyphyllin VI, polyphyllin VII, dioscin, gracillin and IS were m/z 899.5>853.4, 1059.5>1013.5, 783.4>737.4, 1075.5>1029.5, 913.5>867.4, 929.5>883.4 and 819.5>783.4, respectively. The intra- and inter-day precisions (RSD%) were less than 13% and the average extraction recoveries ranged from 85% to 97.0% for each analyte. Six steroidal saponins were proved to be stable during sample storage, preparation and analytical procedures. The established method was employed for simultaneous quantification and successfully used for the first time for the pharmacokinetics evaluation of the six main compounds after intragastric administration of P. polyphylla extract in Sprague–Dawley rats.

English

Chonglou is a well known traditional Chinese medicine for anti-cancer and traumatic injuries. The major sources of Chonglou are from genus Paris. The present investigation aimed to determine contents of active steroidal saponins in Paris plants collected from different locations of Southwest China. The results showed that polyphyllin I and polyphyllin VII could be detected in almost all samples. However, polyphyllin II and polyphyllin VI were not detected in the research. The results indicated that besides Paris polyphylla var. yunnanensis and P.polyphylla var. chinensis, used as medicine traditionally, other Paris species in Southwest China also have medicinal value. Ranges of bioactive compounds in P. polyphylla var. yunnanensis from different parts of the plants were wide. Coefficient of variation of polyphyllin I, polyphyllin II, polyphyllin VII, and total steroidal saponins were 53.4132, 43.9341, 88.2673, and 60.9082%, respectively. Varieties of compounds type and contents in Paris plants would be related to habitats.   Key words: Paris genus, polyphyllin, variation, high performance liquid chromatography (HPLC), Southwest China.

Simultaneous determination and pharmacokinetic study of polyphyllin I, polyphyllin II, polyphyllin VI and polyphyllin VII in beagle dog plasma after oral administration of Rhizoma Paridis extracts by LC‐MS‐MS

For the first time, a rapid and specific LC-MS-MS method has been developed for the analysis of polyphyllin I, polyphyllin II, polyphyllin VI and polyphyllin VII in beagle dog plasma. The method was applied to study the pharmacokinetics of Rhizoma Paridis extracts containing polyphyllin I, polyphyllin II, polyphyllin VI and polyphyllin VII. The analysis was carried out on an Agilent Zorbax XDB-C(18) reversed-phase column (100 × 2.1 mm, 1.8 µm) by isocratic elution with acetonitrile and water (50:50, v/v). The flow rate was 0.25 mL/min. All analytes including internal standards were monitored by selected reaction monitoring with an electrospray ionization source. Linear responses were obtained for polyphyllin I, polyphyllin II, polyphyllin VI and polyphyllin VII ranging from 10 to 5000 ng/mL. The intra-and inter-day precisions (RSDs) were less than 6.66 and 9.15%. The extraction recovery ranged from 95.53 to 104.21% with RSD less than 8.69%. Stability studies showed that polyphyllin I, polyphyllin II, polyphyllin VI and polyphyllin VII were stable in preparation and analytical process. The validated method was successfully used to determine the concentration-time profiles of polyphyllin I, polyphyllin II, polyphyllin VI and polyphyllin VII.

The Microbiological Transformation of Steroidal Saponins by Curvularia lunata.

Abstract The microbiological transformation of polyphyllin I (compound I ), polyphyllin III (compound II ), polyphyllin V (compound III ) and polyphyllin VI (compound IV ) by Curvularia lunata into their corresponding subsaponins, for example, diosgenin-3- O -α- l -arabinofuranosyl (1→4)-β- d -glucopyranoside (compound V ), diosgenin-3- O -α- l -rhamnopyranosyl (1→4)-β- d -glucopyranoside (compound VI ), diosgenin-3- O -β- d -glucopyranoside (compound VII ) and pennogenin-3- O -β- d -glucopyranoside (compound VIII ), were studied in this paper. Curvularia lunata is able to hydrolyze terminal rhamnosyls that are linked by 1→2 C– bond to sugar residues of steroidal saponins at C-3 position with high activity and regioselectivity.

The microbiological transformation of steroidal saponins by Curvularia lunata

The microbiological transformation of polyphyllin I (compound I), polyphyllin III (compound II), polyphyllin V (compound III) and polyphyllin VI (compound IV) by Curvularia lunata into their corresponding subsaponins, for example, diosgenin-3-O-α-l-arabinofuranosyl (1→4)-β-d-glucopyranoside (compound V), diosgenin-3-O-α-l-rhamnopyranosyl (1→4)-β-d-glucopyranoside (compound VI), diosgenin-3-O-β-d-glucopyranoside (compound VII) and pennogenin-3-O-β-d-glucopyranoside (compound VIII), were studied in this paper. Curvularia lunata is able to hydrolyze terminal rhamnosyls that are linked by 1→2 C– bond to sugar residues of steroidal saponins at C-3 position with high activity and regioselectivity.