Pharmacodynamic evaluation of Polygonum capitatum nanofiber functional dressing / 中国药房

OBJECTIVE To evaluate the efficacy of the functional dressing of Polygonum capitatum nanofibers （P-PVP-PCL）. METHODS P-PVP-PCL were prepared by electrospinning technology， and the microstructure of P-PVP-PCL was observed. The antibacterial activity and antioxidant activity of P-PVP-PCL and its effects on the survival rate， adhesion and migration rate of mouse fibroblast L929 cells were investigated. The effects of medical gauze dressing， blank nanofiber dressing （PVP-PCL） and P- PVP-PCL on the healing rate of the wound were investigated by establishing the back skin wound model of rats. The pathological changes of the wound tissue and collagen fiber deposition were observed， as well as the number of platelet endothelial cell adhesion molecule-1 （CD31） positive blood vessels and the expression of transforming growth factor-β （TGF-β） protein in wound tissue. RESULTS P-PVP-PCL had a smooth surface and a double-layer structure at the cross-section. The inhibition rates of P-PVP-PCL against Staphylococcus aureus and Escherichia coli were （98.88±0.66）% and （94.75±1.41）% ， respectively. The antioxidant activity of P-PVP-PCL was （83.69±1.56）%， and the cell activity of the P-PVP-PCL group was significantly higher than those of the control group and PVP-PCL group （P＜0.05）. Compared with medical gauze dressings， P-PVP-PCL was more conducive to L929 cell adhesion； at 48 hours， the cell scratches in this group had basically healed. Compared with the medical gauze dressing group， the wound healing rates of the PVP-PCL group and the P-PVP-PCL group were significantly increased （P＜0.05）. On the 14th day of intervention， the wounds in the P-PVP-PCL group had basically healed， there was no dermal necrosis in the wound tissue， and the collagen fibers were arranged relatively neatly and the density was relatively uniform. The number of CD31 positive blood vessels and the expression of TGF-β protein showed a downward trend compared with the 7th day of intervention， and the number of CD31 positive blood vessels was significantly lower than those of the medical gauze dressing group and PVP-PCL group （P＜0.05）， but the protein expression of TGF- β was significantly higher than those of the medical gauze dressing group and the PVP-PCL group （P＜0.05）. CONCLUSIONS P-PVP-PCL has good antibacterial and antioxidant activity in E-mail：444096585@qq.com vitro， and can promote the proliferation， adhesion and migration of L929 cells. It can promote wound healing of rats in vivo.

Mechanism of Polygonum capitatum in treatment of Helicobacter Pylori associated gastritis by network pharmacology, microarray data analysis and molecular docking / 中国药理学通报

Aim To explore the mechanism of Polygonum capitatum(PC)in the treatment of Helicobacter Pylori associated gastritis(HAG). Methods The databases were used to identify the target of PC active compounds and HAG-related genes,and the intersection was taken to obtain the potential targets of PC treatment of HAG. The interaction network diagram of “drug-active compound-target-disease” and the protein-protein interaction(PPI)network of potential target protein interaction in HAG treated by PC were constructed by software Cytoscape 3.6.0. The important nodes in the network were screened by several topological indexes,and the GO and KEGG enrichment were analyzed by STRING database to obtain the potential signaling pathway of PC in the treatment of HAG. The binding ability of PC active components with key target proteins was observed by molecular docking method. On this basis,the related targets of PC in the treatment of HAG were verified in vivo and in vitro experiments. Results The PC active compounds and targets were identified through the database,and the “drug-active compound-target-disease” network diagram and the PPI network of potential target proteins were constructed. Combined with several topological indexes,the PPI network of potential target-protein interaction was analyzed,and 52 hub genes were screened. Further bioinformatics analysis and high-throughput sequencing revealed that PC exerted an effect on HAG through the Akt/NF-κB/NLRP3 pathway. Based on this,it was found that PC could reduce IL-18 and IL-1β in HAG GES-1 cells and HAG SD rats,up-regulate Akt and its phosphorylation level and reduce NF-κB expression,inhibit the activation of NLRP3 inflammatory body,so as to improve HAG inflammatory response. Conclusions PC could exert a therapeutic effect on HAG by activating Akt and its phosphorylation level,and inhibiting the expression of NF-κB and NLRP3 inflammasome related factors. This study provides a theoretical basis for explaining the mechanism of PC in the treatment of HAG.

Lipidomic investigation of the protective effects of Polygonum perfoliatum against chemical liver injury in mice / 中西医结合学报

OBJECTIVE@#Recent investigations have demonstrated that Polygonum perfoliatum L. can protect against chemical liver injury, but the mechanism behind its efficacy is still unclear. Therefore, we studied the pharmacological mechanism at work in P. perfoliatum protection against chemical liver injury.@*METHODS@#To evaluate the activity of P. perfoliatum against chemical liver injury, levels of alanine transaminase, lactic dehydrogenase, aspartate transaminase, superoxide dismutase, glutathione peroxidase and malondialdehyde were measured, alongside histological assessments of the liver, heart and kidney tissue. A nontargeted lipidomics strategy based on ultra-performance liquid chromatography quadrupole-orbitrap high-resolution mass spectrometry method was used to obtain the lipid profiles of mice with chemical liver injury and following treatment with P. perfoliatum; these profiles were used to understand the possible mechanisms behind P. perfoliatum's protective activity.@*RESULTS@#Lipidomic studies indicated that P. perfoliatum protected against chemical liver injury, and the results were consistent between histological and physiological analyses. By comparing the profiles of liver lipids in model and control mice, we found that the levels of 89 lipids were significantly changed. In animals receiving P. perfoliatum treatment, the levels of 8 lipids were significantly improved, relative to the model animals. The results showed that P. perfoliatum extract could effectively reverse the chemical liver injury and significantly improve the abnormal liver lipid metabolism of mice with chemical liver injury, especially glycerophospholipid metabolism.@*CONCLUSION@#Regulation of enzyme activity related to the glycerophospholipid metabolism pathway may be involved in the mechanism of P. perfoliatum's protection against liver injury. Please cite this article as: Peng L, Chen HG, Zhou X. Lipidomic investigation of the protective effects of Polygonum perfoliatum against chemical liver injury in mice. J Integr Med. 2023; 21(3): 289-301.

Evaluación biológica in vitro del extracto etanólico de Kaa tái, Polygonum punctatum Elliot. (POLYGONACEAE)

En Paraguay, así como en las Américas, la leishmaniasis constituye una de las problemáticas de salud pública importante, debido a su complejidad tanto epidemiológica, clínica y biológica, afectando especialmente a los más pobres y en los países en vía de desarrollo. Polygonum punctatum Elliot. pertenece a la familia Polygonaceae, la medicina popular le atribuye varias propiedades, por medio de estudios anteriores se confirma que diferentes extractos de esta planta presentan actividad antibacteriana, antiinflamatoria y antifúngica. En este trabajo, se evaluó la actividad citotóxica del extracto etanólico de Polygonum punctatum de la familia Polygonaceae en concentraciones de 100, 50 y 25 µg/ml en células de macrófagos de ratones, en los cuales no se encontró actividad citotóxica. Además, se evaluó la actividad leishmanicida por medio de estudios experimentales in vitro a concentraciones de 100, 75, 50, 25, 20 y 10 µg/ml frente a tres cepas de parásitos: Leishmania infantum, L. amazonensis y L. braziliensis, en los cuales se observó una actividad de 58 y 56% de lisis de parásitos con el extracto de 100µg/ml frente a las cepas de L. braziliensis y L. infantum, respectivamente y 51% para L. amazonensis. Estos resultados son prometedores, y aportan una base para el desarrollo y búsqueda de nuevos tratamientos para la leishmaniasis, sin embargo, son necesarios estudios posteriores en cuanto a aislamiento e identificación del compuesto y evaluación en modelos animales in vivo.

In Paraguay, as well as in the Americas, leishmaniasis constitutes one of the important public health problems, due to its epidemiological, clinical, and biological complexity, especially affecting the poorest and developing countries. Polygonum punctatum Elliot belongs to the Polygonaceae family, and popular medicine attributes several properties to it. Previous studies confirmed that different extracts of this plant have antibacterial, anti-inflammatory and antifungal activity. In this work, the cytotoxic activity of the ethanolic extract of Polygonum punctatum was evaluated at concentrations of 100, 50 and 25 µg/ml in mouse macrophage cells, in which no cytotoxic activity was found. In addition, the leishmanicidal activity was evaluated through experimental in vitro studies at concentrations 100, 75, 50, 25, 20 and 10 µg/ml against three strains of parasites: Leishmania infantum, L. amazonensis and L. braziliensis. Activities of 58 and 56% of parasite lysis were observed with the 100 µg/ml extract against strains of L. braziliensis and L. infantum, respectively, and 51% for L. amazonensis. These results are promising and provide a basis for the development and search of a new treatment for leishmaniasis. However, further studies are necessary regarding the isolation and identification of the compound and its evaluation in in vivo animal models.

Discovery of pulmonary fibrosis inhibitor targeting TGF-β RI in Polygonum cuspidatum by high resolution mass spectrometry with in silico strategy / 药物分析学报

Pulmonary fibrosis(PF)is an irreversible lung disease that is characterized by excessive scar tissue with a poor median survival rate of 2-3 years.The inhibition of transforming growth factor-β receptor type-Ⅰ(TGF-β RI)by an appropriate drug may provide a promising strategy for the treatment of this disease.Polygonum cuspidatum(PC)is a well-known traditional Chinese herbal medicine which has an anti-PF effect.Accordingly,a combination of high resolution mass spectrometry with an in silico strategy was developed as a new method to search for potential chemical ingredients of PC that target the TGF-β RI.Based on this strategy,a total of 24 ingredients were identified.Then,absorption,distribution,meta-bolism,and excretion(ADME)-related properties were subsequently predicted to exclude compounds with potentially undesirable pharmacokinetics behaviour.Molecular docking studies on TGF-β RI were adopted to discover new PF inhibitors.Eventually,a compound that exists in PC known as resveratrol was proven to have excellent biological activity on TGF-β RI,with an ICso of 2.211 μM in vitro.Furthermore,the complex formed through molecular docking was tested via molecular dynamics simulations,which revealed that resveratrol had strong interactions with residues of TGF-β RI.This study revealed that resveratrol has significant potential as a treatment for PF due to its ability to target TGF-β RI.In addition,this research demonstrated the exploration of natural products with excellent biological activities toward specific targets via high resolution mass spectrometry in combination with in silico technology is a promising strategy for the discovery of novel drugs.

Comparison of the fingerprints and in vitro antioxidant activity of decoction pieces of Polygonum cuspidatum by integrated processing and traditional processing / 中国药房

OBJECTIVE To compare t he diff erences of the fingerprint and in vitro antioxidant activity between decoction pieces of Polygonum cuspidatum by integrated technology of habitat processing and processing （short for IPDP ）and traditional processing decoction pieces （short for TPDP ）. METHODS Ten batches of IPDP and ten batches of TPDP were prepared by integrated technology and traditional technology ，respectively. HPLC method was used to establish and compare the fingerprints of IPDP and TPDP. The scavenging rates of DPPH free radical ，ABTS free radical ，superoxide free radical and hydroxyl free radical and reducing activity of Fe 3+ were detected for IPDP and TPDP. In vitro antioxidant activities were compared between IPDP and TPDP. RESULTS There were 11 common peaks in the fingerprints of IPDP and TPDP ，among which 17 came from IPDP and 13 came from TPDP. The peak heights of peak 6（polydatin）and peak 15（emodin-8-O-β-D-glucoside）in IPDP were significantly higher than those in the TPDP ，and the peak heights of peak 13（resveratrol），peak 17（emodin）and peak 19（physcion）in the TPDP were significantly higher than those in the IPDP. The results of in vitro antioxidant test showed that IPDP and TPDP had a certain scavenging capacity on DPPH free radical ，ABTS free radical ，superoxide free radical and hydroxyl free radicals ，and also had a certain reducing capacity on Fe 3+. CONCLUSIONS The integrated processing technology of P. cuspidatum has a good retention effect on the glycosides in P. cuspidatum ，and the in vitro antioxidant activity of IPDP is stronger than that of TPDP.

Signaling pathways involved in the active components of Polygonum cuspidatum in treatment of nonalcoholic fatty liver disease and their interaction / 临床肝胆病杂志

The pathogenesis of nonalcoholic fatty liver disease (NAFLD) remains unclear, and currently no effective drugs have been approved for the treatment of NAFLD. Polygonum cuspidatum is a natural traditional Chinese medicine with a long history of application, and studies have shown that it plays an important role in the treatment of NAFLD. This article summarizes related research findings in the active components of Polygonum cuspidatum applied in the treatment of NAFLD, and it is found that the active components of Polygonum cuspidatum can improve insulin resistance, exert an anti-oxidative stress effect, regulate lipid metabolism, improve endoplasmic reticulum stress, and alleviate inflammatory infiltration by regulating the signaling pathways including Nrf2, AMPK, NF-κB, SIRT1, and PPARα, thereby exerting a preventive and therapeutic effect on NAFLD, so as to provide a basis and ideas for developing drugs for NAFLD and exploring related mechanisms.

Identification of Escherichia coli β-glucuronidase inhibitors from Polygonum cuspidatum Siebold & Zucc

Abstract Gut bacterial β-glucuronidase (GUS) can reactivate xenobiotics that exert enterohepatic circulation- triggered gastrointestinal tract toxicity. GUS inhibitors can alleviate drug-induced enteropathy and improve treatment outcomes. We evaluated the inhibitory effect of Polygonum cuspidatum Siebold & Zucc. and its major constituents against Escherichia coli GUS (EcGUS), and characterized the inhibitory mechanism of each of the components. Trans-resveratrol 4'-O-β-D-glucopyranoside (HZ-1) and (-)-epicatechin gallate (HZ-2) isolated from P. cuspidatum were identified as the key components and potent inhibitors. These two components displayed strong to moderate inhibitory effects on EcGUS, with Ki values of 9.95 and 1.95 μM, respectively. Results from molecular docking indicated that HZ-1 and HZ-2 could interact with the key residues Asp163, Ser360, Ile 363, Glu413, Glu504, and Lys 568 of EcGUS via hydrogen bonding. Our findings demonstrate the inhibitory effect of P. cuspidatum and its two components on EcGUS, which supported the further evaluation and development of P. cuspidatum and its two active components as novel candidates for alleviating drug-induced damage in the mammalian gut.

Study on Improvement Effects of Polygonum orientale Flower Extract on Hypoxia-reoxygenation Injury of H 9c2 Cardiomyocytes / 中国药房

OBJECTIVE：To study the improvement effects and mechanism of Polygonum orientale flower extract on hypoxia- reoxygenation injury of H 9c2 cardiomyocytes. METHODS ：H9c2 cardiomyocytes were divided into normal control group ，model group and low- ，medium- and high-concentrations groups of P. orientale flower extract （20，40，80 μg/mL）. Except for normal control group ，other groups were given 800 μmol/L CoCl2 to induce hypoxia-reoxygenation injury model. Cell apoptosis was observed. The levels of Ca 2+（in cytoplasm ），mitochondrial membrane potential （MMP），ATP enzyme （Na+-K+-ATP enzyme ，Ca2+-Mg2+-ATP enzyme） activities， the ratio of cytochrome c （Cyto c ）， protein in cytosol to mitochondria ，phosphorylation levels of reperfusion injury salvage kinase （RISK） signaling pathwayrelated protein [protein kinase B （Akt）and extracellular signal regulated kinase 1/2（ERK1/2）] as well as protein expression of HIF- 1 α were detected respectively. In addition，the cells were divided into normal control group ，model group and P. orientale flower extract group （80 μ g/mL），PI3K inhibitor LY294002+CoCl2 group（15 μmol/L LY294002+80 μmol/L ，LY294002+P. orientale flower extract group （15 μmol/L LY294002+80 μg/mL P. orientale flower extract ），MEK inhibitor PD98059+CoCl2 group（25 μmol/L PD98059+800 μmol/L CoCl2），PD98059+P. orientale flower extract group （25 μmol/L PD98059+80 μg/mL P. orientale flower extract ）. After cultured by the same method ，the phosphorylation levels of Akt protein and ERK1/2 protein in the cells were measured to verify the activation of P. orientale flower extract to RISK signaling pathway. RESULTS：Compared with model group ，nuclear pyknosis and the number of apoptotic bodies were reduced in different concentrations groups of P. orientale flower extract. ROS level ，Ca2+ level（except for low-concentration group ），MMP，ratio of Cyto c in cytoplasm to Cyto c in mitochondria ，protein expression of HIF- 1α were decreased significantly（P＜0.05 or P＜0.01）； the activity of ATP enzyme （except for the low-concentration group ），Akt protein and ERK 1/2 protein phosphorylation level were significantly increased （P＜0.01）. After treated with PI 3K inhibitor LY 294002 and MEK inhibitor PD 98059，Akt protein and ERK 1/2 protein phosphorylation level in cadiomyocyte were decreased significantly （P＜0.05 or P＜0.01）. CONCLUSIONS ：P. orientale flower extract can improve hypoxia-reoxygenation injury of H 9c2 cardiomyocytes，the mechanism of which may be associated with inhibiting cardiomyocyte apoptosis ，improving ATPase activity ，protecting mitochondria ，regulating RISK signaling pathway related proteins and HIF- 1α protein expression.

The compound cell model-based evaluation for idiosyncratic liver injury of Cis-SG and Trans-SG / 药学学报

In this study, a composite cell model for evaluation of idiosyncratic drug-induced liver injury (IDILI) was established in vitro from the perspective of immune inflammation. And this model was used to evaluate the risk of IDILI for 2,3,5,4'-tetrahydroxy-cis-stilbene-2-O-β-glucoside (Cis-SG) and 2,3,5,4'-tetrahydroxy-trans-stilbene-2-O-β-glucoside (Trans-SG). To determine the low, medium, and high dosage of Cis-SG and Trans-SG, CellTiter-Glo® 3D Cell Viability Assay was used to detect the effects of Cis-SG and Trans-SG on cell viability of HepG2 cells in three dimensional (3D) culture, and MTT assay was used to detect the effects of Cis-SG and Trans-SG on cell viability of THP-1 derived macrophages. THP-1 derived macrophages were incubated by Cis-SG and Trans-SG directly or supernatants from HepG2 cells incubated with them. Enzyme linked immunosorbent assay (ELISA) was used to detect the levels of interleukin-1β (IL-1β) in the supernatants of the THP-1 derived macrophages. Western blot and reverse transcription-polymerase chain reaction (RT-PCR) were used to determine the expression of apoptosis-associated speck-like protein (ASC), Nod-like receptor protein 3 (NLRP3), cysteinyl aspartate specific proteinase-1 (caspase-1), and IL-1β in THP-1 derived macrophages. The results showed that there was no effect on the secretion of IL-1β in THP-1 derived macrophages incubated by Cis-SG and Trans-SG directly. However, the secretion of IL-1β, the protein and mRNA expression of ASC, NLRP3, caspase-1, and IL-1β significantly increased in THP-1 derived macrophages incubated by supernatants from HepG2 cells incubated with 1, 5, and 25 μmol·L-1 Cis-SG or 25 μmol·L-1 Trans-SG. In summary, the composite cell model for evaluation of IDILI established in vitro has been successfully applied in testing Cis-SG and Trans-SG. This composite cell model is helpful to evaluate and screen drugs with IDILI risk in vitro preliminarily, which provides methods for predicting and solving the idiosyncratic liver toxicity of drugs.

Two new sucrose cinnamates from Polygonum lapathifolium var. salicifolium / 中国中药杂志

Two new sucrose cinnamates(1 and 2) along with nine known compounds(3-11) were isolated from ethanol extract of Polygonum lapathifolium var. salicifolium by silica gel column chromatography, ODS column chromatography and semi-preparative HPLC. Their structures were elucidated by extensive spectroscopic methods including 1 D-and 2 D-NMR experiments, as well as HR-ESI-MS analysis. Eleven compounds(7 sucrose cinnamates, 3 phenylpropanoids and 1 lactone) were obtained and their structures were identified as(1,3-O-di-p-coumaroyl)-β-D-fructofuranosyl-(2→1)-α-D-glucopyranoside(1),(1,3-O-di-p-coumaroyl)-β-D-fructofuranosyl-(2→1)-(6-O-acetyl)-α-D-glucopyranoside(2),(3-O-feruloyl)-β-D-fructofuranosyl-(2→1)-(6-O-p-coumaroyl)-α-D-glucopyranoside(3), hydropiperoside(4), vanicoside C(5),(1,3-O-di-p-coumaroyl)-β-D-fructofuranosyl-(2→1)-(6-O-feruloyl)-α-D-glucopyranoside(6), vanicoside B(7),trans-p-hydroxycinnamic acid methyl ester(8), trans-p-hydroxycinnamic acid ethyl ester(9), methyl ferulate(10) and dimethoxydimethylphthalide(11), respectively. Compounds 1 and 2 were two new sucrose cinnamates, and compounds 1-11 were isolated from this plant for the first time. The antioxidant activities of the isolated compounds 1-9 were investigated by an oxygen radical absorbance capacity(ORAC) assay, and all nine compounds were found to show strong antioxidant activities. Among them, compound 6(10 μmol·L~(-1)) was the supreme one in antioxidant activities, with its ORAC value equivalent to(1.60±0.05) times of 50 μmol·L~(-1) Trolox.

Intestinal absorption characteristics of Polygonum orientale extract in normal and isoproterenol-induced myocardial ischemia model rats via everted intestinal sac models / 中国中药杂志

The present study is to investigate the absorption characteristics of the main components in Polygonum orientale extract in normal and isoproterenol-induced myocardial ischemia model rats with everted intestinal sac models. Intestinal sac fluid samples were collected in different part of intestine(duodenum, jejunum, ileum, colon) at different time after administration of different concentration of P. orientale extract(5.0,10.0, 20.0 mg·mL~(-1)). An UPLC-TQD method was employed for the determination of six components including orientin, isoorientin, vitexin, protocatechuic acid, kaempferol-3-O-β-D-glucoside and quercitrin in the intestinal sac samples. The absorption rate and cumulative absorption were calculated to analyze the intestinal absorption characteristics of six components in normal and myocardial ischemia model rats. The P-glycoprotein(P-gp) inhibitor was applied to investigate influence of intestinal absorption of six components in P. orientale extract. The results showed that the main absorption sites were concentrated on the duodenum at low concentration, while they were the colon at the medium concentration and the ileum at high concentration in control groups. In the condition of myocardial ischemia model, the main absorption sites focus on the ileum and jejunum at low concentration; the main absorption sites were in the ileum at the medium concentration and main absorption sites were the duodenum and ileum at high concentration. Compared with the normal group, the absorption rate and cumulative absorption of the six components significantly decreased in the model group. P-gp inhibitor markedly increased the absorption rate and cumulative absorption of six components in the model group, inferring that the 6 components may be the substrates of P-gp, and the mechanism needs further study. In this study, it is revealed that the six components of P. orientale extract can be absorbed into the intestinal sac, and it is an effective method to assess the intestinal absorption characteristics of P. orientale extract through everted intestinal sac model, providing data support for the clinical application and further development of P. orientale.

Reference gene screening for Real-time quantitative PCR in Polygonum multiflorum / 中国中药杂志

To select suitable references gene of Polygonum multiflorum for gene expression analysis in different tissues, five candidate reference genes like Actin,GAPDH,SAND,PP2A,TIP41 were selected from the transcriptome data of P. multiflorum, then the specific primers were designed. The expression stability of the five reference genes in different tissues of P. multiflorum was analyzed by Real-time quantitative PCR through avilable analysis methods such as geNorm, NormFinder, BestKeeper, Delta CT and RefFinder, to ensure the reliability of the analysis results. The results showed that there were significant differences in the expression levels and stability of candidate genes in different tissues of P. multiflorum. Ct distribution analysis of the expression levels of candidate genes showed that the expression levels of Actin and GAPDH genes were relatively high in different tissues, while the expression levels of SAND, PP2A and TIP41 were lower. The stability of each candidate gene was analyzed by different methods. The results of geNorm analysis showed that the expression of PP2A and GAPDH was the most stable, the expression stability of SAND was the worst, the stability of PP2A was the highest in both NormFinder and Delta CT, the stability of SAND was the lowest, and the stability of Actin was the most stable in BestKeeper analysis. Through the comprehensive evaluation and analysis of the stability of candidate genes by RefFinder, it is concluded that the stability of PP2A gene is the highest, followed by GAPDH, Actin, TIP41, SAND, and SAND gene is the worst. Therefore, the PP2A gene is an ideal reference gene for the analysis of gene expression in different tissues of P. multiflorum.

Quality Evaluation of Polygonum cuspidatum by UPLC Fingerprint and QAMS / 中国药房

OBJECTIVE：To establish the UPLC fingerprint of Polygonum cuspidatum ，and to determine the contents of four active ingredients and to provide reference for the quality evaluation of P. cuspidatum . METHODS ：The determination was performed on Waters BEH C 18 column（100 mm×2.1 mm，1.7 μm）with mobile phase consisted of acetonitrile- 0.2％ formic acid （gradient elution ）at flow rate of 0.4 mL/min. The column temperature was 40 ℃，and detection wavelength was 290 nm. The sample size was 1 μL. The fingerprints were evaluated by similarity calculation，cluster analysis and orthogonal partial least square discriminant analysis （OPLS-DA）. Using polydatin as internal standard ，relative calibration factors of resveratrol ，emodin-8-O- β-D-glucoside and emodin were determined to develop a method of QAMS. The contents of 4 above components in 15 batches of P. cuspidatum were calculated by relative calibration factors. The results of QAMS were compared with those of external standard. RESULTS：UPLC fingerprints of 15 batches of P. cuspidatum were established ，and 12 common peaks were confirmed. Five components were identified ，i.e. polydatin ，resveratrol，emodin-8-O-β-D-glucoside，emodin，emodin methyl ether. The fingerprint similarity of 15 batches of P. cuspidatum was in the range of 0.865-0.976. According to cluster analysis ，15 batches of P. cuspidatum were classified into 4 categories，showing certain regularity of origin. Seven markers were identified by OPLS-DA method. The order of difference significance was peak 7＞emodin-8-O-β-D-glucoside＞resveratrol＞peak 8＞polydatin＞peak 1＞ peak 10. The relative deviation among the contents of resveratrol ，emodin-8-O-β-D-glucoside and emodin in 15 batches of P. cuspidatum determined by QAMS and external standard method was less than 5.0％，indicating that there was no significant difference between the two methods. CONCLUSIONS ：UPLC fingerprint combined with QAMS method is convenient and reliable for the quality evaluation of P. cuspidatum ；the quality of P. cuspidatum produced in Chongqing and Anhui province is better.

Pharmacokinetic study of Polygonum orientale extract in H9c2 cells by UPLC-MS/MS / 中国中药杂志

A detection method of ultra-performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS) was established to detect concentrations of isoorientin, orientin, quercetin, vitexin and kaempferol-3-O-β-D-glucoside in H9 c2 cells and applied to the pharmacokinetic study of Polygonum orientale extract in the cells. H9 c2 cells were treated with 100 μg·mL~(-1) P. orientale extract and then they and the corresponding nuclei, mitochondria and Golgi bodies were collected at the set time. After protein precipitation, UPLC-MS/MS was used to determine concentrations of isoorientin, orientin, quercetin, vitexin and kaempferol-3-O-β-D-glucoside in the whole cells and subcellular structures. Also, related pharmacokinetic parameters were calculated. The results showed that the peak time was 8 h for all these components. Orientin, vitexin, quercetin and isoorientin have high affinities to nuclei and mitochondria, while the affinity of kaempferol-3-O-β-D-glucoside is higher with mitochondria compared to nuclei. It is suggested that these chemical components of P. orientale may mainly act on nuclei or mitochondria to exert pharmacological effects of protecting cardiomyocytes.

Screening of plant growth-promoting rhizobacteria and its effect on seed germination of Polygonum multiflorum / 中国中药杂志

In this study, the rhizobacteria and actinomycetes of Polygonum multiflorum were screened for the strains with indole acetic acid(IAA)-producing capacity by Salkowski method, the siderophore-producing strains by Chrome Azurol S(CAS) assay, and the strains with inorganic phosphorus-solubilizing capacity by PKO inorganic phosphorus medium. The strains were identified by morphological identification, physiological and biochemical characteristics, and 16 S rDNA sequences. Furthermore, the effect of growth-promoting strains on the seed germination and development of P. multiflorum was tested. The results showed that among 196 strains, two strains F17 and F42 were found to be capable of producing IAA and siderophore and solubilizing inorganic phosphorus simulta-neously. For F17 and F42, the results are listed below: 38.65 and 33.64 mg·L~(-1) for IAA production, 0.85 and 0.49 for siderophore-producing capacities(A_s/A_r), and 1.35 and 1.70 for inorganic phosphorus-solubilizing capacities(D/d), respectively. Comprehensive analysis revealed that strains F17 and F42 were identified as Pseudochrobactrum asacharolyticum and Bacillus aryabhattai, respectively, and both could significantly promote the seed germination of P. multiflorum.

Hepatotoxicity of Main Components of Polygonum multiflorum and Its Effects on Drug Metabolism Enzyme / 中国药房

OBJECTIVE：To study the hepatotoxicity of main components of Polygonum multiflorum ，and investigate its toxic mechanism based on metabolic enzymes. METHODS ：ADMETlab 2.0 platform was used to forecast the toxic or carcinogenic effects of emodin ，physcion，rhein，stilbene glycoside and gallic acid on liver ，skin and heart. The effects of those components on cytochrome P 450 enzyme system （CYP1A2，CYP2C9，CYP2C19，CYP2D6，CYP3A4）were evaluated. The effects of different concentrations of emodin ，rhein，stilbene glycoside and gallic acid （10，20，40，80 μmol/L）on the survival rate of normal hepatocyte L 02 were detected. The effects of major components of P. multiflorum on the activity of UGT 1A1 enzyme were studied by in vitro reaction system ，using bilirubin as substrate. RESULTS ：Main components of P. multiflorum ，ie. emodin ，physcion， rhein and gallic acid ，showed strong toxic effects on the liver ，while stilbene glycosides possessed weak toxic effects on the liver. Emodin and physcion had strong inhibitory effects on CYP 1A2 and medium inhibitory effects on CYP 2C9，CYP2D6 and CYP3A4；rhein showed medium inhibitory effects on CYP 1A2 and CYP 2C9，while stilbene glycoside and gallic acid possessed weak inhibitory effects on the above enzymes. Emodin （40，80 μmol/L）and gallic acid （40，80 μmol/L）could significantly reduce the survival rate of L 02 cells（P＜0.01）. The inhibition rate of 5，10，20，40，80 μmol/L emodin and gallic acid（except for 5 μmol/L emodin）on UGT 1A1 enzyme increased significantly （P＜0.01），and the inhibition effect of emodin on UGT 1A1 enzyme was reversible competitive inhibition. CONCLUSIONS ：The main components of P. multiflorum ，ie. emodin ，rhein and physcion ， are hepatotoxic ；the mechanism of it may be associated with inhibiting the activity of CYP 1A2 and CYP 2C9 and competitively blocking rate-limiting enzyme UGT 1A1 in the process of bilirubin metabolism.

Polygonum hydropiper extract attenuates ethanol-induced gastric damage through antioxidant and anti-inflammatory pathways

The present study was conducted to investigate the underlying mechanisms and effective components of Polygonum hydropiper in ethanol-induced acute gastric mucosal lesions. The ethanol extract was purified on an AB-8 macroporous resin column and eluted with 60% ethanol and was then injected into the HPLC system for quantitative analysis. Sprague-Dawley rats were orally pretreated with P. hydropiper extract (PHLE; 50, 100, and 200 mg/kg) for 5 days and then absolute ethanol was administered to induce gastric mucosal damage. One hour after ethanol ingestion, the rats were euthanized and stomach samples were collected for biochemical analysis. Antioxidant enzymes and anti-inflammatory cytokines were quantified. Western blotting was used to detect the expression levels of proteins. Cell proliferation was assayed by CCK-8 assays. The proportion of total flavonoids in the final extract of P. hydropiper was 50.05%, which contained three major bioactive flavonoid constituents, including rutin, quercitrin, and quercetin. PHLE significantly increased cell viability and effectively protected human gastric epithelial cells-1 against alcohol-induced damage in vitro. PHLE pretreatment attenuated gastric mucosal injuries in a dose-dependent manner in rats, and increased the activity of superoxide dismutase, glutathione peroxidase, and glutathione, and decreased the levels of malondialdehyde in gastric tissue. Pretreatment with PHLE also reduced the generation of the pro-inflammatory cytokines tumor necrosis factor-α and interleukin-1β in gastric tissue by downregulating the expression of nuclear factor-kappa B. PHLE exerted protective effects against gastric injury through antioxidant and anti-inflammatory pathways. Flavonoids might be the main effective components of P. hydropiper against gastric mucosal injury.

Different Polarity Extracts of Polygonum minus towards Cytotoxic Activities against Colon Cancer Cell Lines (HT-29, HCT-116, CT-26)

Aims: To investigate the cytotoxic properties of different polarity solvents of Polygonum minusextracts towards colon cancer cell lines, HT-29, HCT-116 and CT-26.Study Design:Experimental study. Place andDuration of Study:Central Laboratory, Tissue Culture Laboratory, Universiti Sultan Zainal Abidin, Terengganu between September 2019 until December 2019.Methodology:The different polarity solvents of P. minusextracts had been led to tetrazolium salt reduction (MTT) assay and an inhibition concentration of 50 (IC50) value for their cytotoxic potential against colon cancer cells. Then, cell morphology observation and fluorescence double staining of treatment cells were determined using a light inverted microscope and acridine orange/propidium iodide staining.Results:The results indicated that an extraction yields aligned from 0.01% for acetone and ethyl acetate to 0.45% for aqueous solution with decreasing order of aqueous solution > 70% aqueous ethanol > 50% aqueous ethanol > methanol > ethanol > acetone and ethyl acetate. Meanwhile, the ethyl acetate extract showed a higher cytotoxic effect at IC50values of 7.00 ± 0.06 μg/mL and 7.00 ± 0.30 μg/mL towards the HCT-116 and CT-26 cells; and 50% aqueous ethanol towards HT-29 cells (24.00 ± 0.01 μg/mL). The different solvent extracts of P. minusinduced cytotoxic effects on the treated cell lines by altering their normal cell morphology and cell membrane integrity (except for acetone extract). Conclusion:Therefore, the use of different polarity solvent extracts of P. minusas an anti-cancer agent is promising more on ethyl acetate and warrants further investigation

Chemical Constituents from Polygonum cuspidatum / 中国药学杂志

OBJECTIVE: To study the chemical constituents from Polygonum cuspidatum. METHODS: The petroleum ether-ethyl acetate solvents at different ratios of 9:1, 7:1, 5:1, 3:1 and 1:1 were used as gradient eluents for silica gel column chromatography, and samples collected were further isolated and purified by preparative high performance liquid chromatography. Their structures were elucidated by physico-chemical constants and spectroscopic methods. RESULTS: Eight compounds were obtained and identified as 22(Z)-ergosterol-4,6,8,22-tetraene-3-one(1), (22E,24R)-stigma-1,4-diene-3-one(2), (E)-ethyl octadecanoic-16-enoic acid ethyl ester(3), oleanolic acid(4), emodin(5), resveratrol(6), trans-resveratrol glycoside(7) and 6'-gallic acid-4-O-D-resveratrol ester(8).CONCLUSION: Compound 1 is a new natural compound (patent No. ZL 2013 10205435.5), compound 2 and 3 are isolated from this plant for the first time.