Mechanism of Polygoni Cuspidati Rhizoma in treating atherosclerosis based on network pharmacology and molecular docking

Background:The incidence and prevalence of atherosclerosis(AS)is increasing every year and has becoming a major health issue of global concern.Polygoni Cuspidati Rhizoma(PCR)is a Chinese herb that is widely used clinically for the treating of AS.However,its pertinent targets and probable mechanisms,still need to be completely explored.Methods:Active compounds and targets for PCR and AS targets were screened using public databases.A“drug-component-disease target”network map was created and analyzed after using the Venn online tool to identify common targets and Cytoscape software to screen drug-disease core targets.Critical targets pathway enrichment analyses are conducted using the Metascape database.Using AutoDock Vina and Pymol software,docking validation and visualization of active components and core targets were carried out.Results:PCR was obtained for ten compounds with 105 AS-related targets.Rhein,quercetin,beta-sitosterol,and luteolin may be drug candidates,and the genes for AKT1,TNF,IL-6,EGFR,TP53,IL-1,RELA,and VEGFA are potential therapeutic targets,according to network analysis.PCR might modulate the AGE/RAGE,PI3K/Akt,IL-17 and NF-ᴋB signaling pathways against the development of AS.Molecular docking indicated that quercetin has high affinity for AKT1 and TNF gene targets.Conclusion:This study provides rare information and scientific basis for further exploration of PC in the treatment of AS.

Two New Glucosides from Radix Polygoni multiflori Preparata

2,3,5,4'-tetrahydroxystilbene-2-O-(2'-O-feruloyl)-beta -D-glucopyranoside, 2,3,5,4'-tetra hydroxystilbene-2-O-(2'-O-p-coumaroyl)-beta -D-glucopyranoside were isolated from Radix Polygoni multiflori Preparata. Structures were elucidated by chemical and spectral evidences.

Inhibitory Effects of Polygoni Multiflori Caulis Extracts on Matrix Metalloproteinase Activities

Matrix metalloproteinases（MMPs） play an essential role in development and tissue remodeling of living organisms. However, the overexpression of MMPs has lead to a series of diseases, such as cancer, arthritis, and atherosclerosis; and inhibition of MMPs may have therapeutic benefits. The discovery of MMP inhibitors from herbal has become a prospective event. We showed that the extract of Polygoni multiflori caulis from ethyl acetate or water（ethyl acetate extract and water extract） can inhibit the activities of MMPs 9, 14, and 16 in a dose-dependent manner and n-butyl alcohol extract of it can also inhibit these MMPs. Furthermore, we found that n-butyl alcohol extract and water extract of it influence the cell viability. These discoveries may contribute to the development of MMP in- hibitors for the therapy of a variety of pathological conditions.

Protective Effects and Action Mechanism of Total Flavonoids of POLYGONI PERFOLIATI HERBA on Rats with Nonalcoholic Steatohepatitis

[Objectives]To study the protective effects and mechanism of POLYGONI PERFOLIATI HERBA on rats with nonalcoholic steatohepatitis( NASH). [Methods] Sixty rats were randomly divided into normal group,model group,metformin group( 0. 5 mg/kg),and high dose,medium dose,and low dose groups of total flavonoids of POLYGONI PERFOLIATI HERBA( 600,300,150 mg/kg). The standard feed was given to the normal group,and the model group and the total flavonoids groups were fed with high-fat diet for 13 weeks to establish the NASH rat model. At the 8 th week,the metformin group and the POLYGONI PERFOLIATI HERBA total flavonoids groups were given with the corresponding drug treatment for 6 weeks,blood was taken from the eyeball to collect liver tissue. Biochemical method was used to determine ALT,AST,TC,TG,HDL-c,LDL-c,SOD,MDA,GSH-Px activity or content in serum and HOMA-IR,and ELISA was used to measure the TNF-α,IL-1β and IL-6 content in liver tissue; Western blotting was used to detect the expression levels of AMPK,p-AMPK and ACC in liver tissue. [Results] Total flavonoids of POLYGONI PERFOLIATI HERBA could significantly decrease the activity or content of ALT,AST and MDA in serum of NASH rats( P < 0. 05,P < 0. 01),and enhance the activity of SOD and GSH-Px( P < 0. 05,P < 0. 01),reduce serum TC,TG,LDL-c levels and insulin resistance index,increase HDL-c levels( P < 0. 05,P < 0. 01); down-regulate liver IL-1β,IL-6,TNF-α,ACC levels and up-regulate p-AMPK expression( P < 0. 05,P < 0. 01). [Conclusions]The total flavonoids of POLYGONI PERFOLIATI HERBA have a good protective effect on NASH rats,and its mechanism may be related to the functions of regulating the lipid metabolism,alleviating insulin resistance,inhibiting oxidative stress,inhibiting inflammatory reaction and regulating AMPK and ACC protein expression.

Radix Polygoni Multiflori Praeparata and Dioscorea Bulbifera Rhizome Decoctions Display Combined Effects Detected by a Three-Probe Drug Cocktail with Substrates of Rat Hepatic Cytochrome P450 Enzymes

Objectives: Radix Polygoni Multiflori Praeparata (RPMP) and Dioscorea Bulbifera Rhizomes (DBR) are used in Chinese herbal medicine and have been frequently reported for adverse reactions on liver. In this research, we aimed to evaluate in vivo effects of RPMP and DBR on rat cytochrome P450 enzymes (CYP1A2, CYP2E1 and CYP3A2) with their respective substrates as probes. Methods: Rats were orally administered RPMP, DBR and RPMP/DBR combination at 12, 10 and (12 + 10) g/kg, respectively, or saline as a control, once daily for 7 days. Thereafter, a cocktail containing 10 mg/kg caffeine, 20 mg/kg chlorzoxazone and 10 mg/kg dapsone was tail vein injected to rats. At defined time points, plasma drug concentrations were simultaneously evaluated by HPLC. Pharmacokinetic parameters simulated by DAS software were used to assess RPMP and/or DBR effects on cytochrome P450 enzymes activity. ANOVA and Dunnett’s test were used for data analysis. Results: Caffeine metabolism was enhanced in RPMP animals and reduced after pretreatment with DBR, but no effect was observed in RPMP/DBR combination group. Chlorzoxazone and dapsone metabolism was enhanced in both RPMP and DBR groups and consequently in combination group. The data suggested that RPMP independently induces rat CYP1A2, CYP2E1 and CYP3A2 activity, while DBR independently inhibits activity of rat CYP1A2 and induces that of CYP2E1 and CYP3A2. RPMP/DBR combination showed no significant benefit compared with the two drugs alone and even showed a neutralized effect in CYP1A2 activity. Conclusions: Caution is needed when RPMP and/or DBR are co-administered with drugs metabolized by human CYP1A2, CYP2E1 and CYP3A4.

Difference Analysis of Secondary Metabolites of Herba Polygoni Chinensis from Guangxi

[Objectives]The differences in secondary metabolites of Herba Polygoni Chinensis from Guangxi were analyzed,in order to find flavonoids which can antagonize liver injury and provide a basis for the quality evaluation of Herba Polygoni Chinensis.[Methods]UPLC-MS was used to analyze the secondary metabolites of Herba Polygoni Chinensis,and emphasis was placed on flavonoids.[Results]Flavonoids were abundant in the secondary metabolites of Herba Polygoni Chinensis.Quercitrin,catechin gallate,epicatechin gallate,hyperoside and so on were screened as the components with high content and antagonistic effect on liver injury.[Conclusions]Quercitrin was the flavonoid compound with the highest content among the three origins.It is stable and has a good protective effect on liver,which is representative.Quercitrin can be considered as the target component to be used as an important index for quality evaluation of Herba Polygoni Chinensis.

Safety Limit Test of Heavy Metals and Pesticide Residues in the Zhuang Ethnic Medcine Herba Polygoni Chinensis from 13 Producing Areas

[Objectives]This study was conducted to determine the contents of such five heavy metals as lead(Pb),cadmium(Cd),arsenic(As),mercury(Hg)and copper(Cu)and the contents of nine kinds of organochlorine pesticide residues including total BHC(α-BHC,β-BHC,γ-BHC,δ-BHC)and total DDT(pp-DDE,pp-DDD,op-DDT,pp-DDT)and pentachloronitrobenzene in the Zhuang medicine Herba Polygoni Chinensis from 13 producing areas.[Methods]The five heavy metals were determined in accordance with the second method of general rule 2341,the inductively coupled plasma mass spectrometry method,in part 4 of the 2020 edition of Chinese Pharmacopoeia;and the nine organochlorines were determined in accordance with the first method of general rule 2341,organochlorine pesticide residue determination method(chromatography),in part 4 of the 2020 edition of Chinese Pharmacopoeia.[Results]In the Herba Polygoni Chinensis from 13 origins,the lead contents were in the range of 0.50-3.28 mg/kg;the cadmium contents were in the range of 0.06-4.21 mg/kg;the arsenic contents were in the range of 0.16-6.9 mg/kg;the copper contents were in the range of 10.5-45.1 mg/kg;the mercury contents were in the range of 0.00-0.003 mg/kg;and the nine organochlorines including the total benzene hexachloride(α-BHC,β-BHC,γ-BHC,δ-BHC),total DDT(pp-DDE,pp-DDD,op-DDT,pp-DDT)and pentachloronitrobenzene were all not detected.Referring to part 4 of the 2020 edition of Chinese Pharmacopoeia,the Cu and As of the Herba Polygoni Chinensis produced in Hezhou exceeded the standards,the Cu of the Herba Polygoni Chinensis produced in Wuzhou exceeded the standard,and the As of the Herba Polygoni Chinensis produced in Yulin City and Guilin City exceeded the standard.[Conclusions]This study provides safety limit test data for the development and utilization of Herba Polygoni Chinensis of the Zhuang medicine.

Expression of a Bacterial Chitinase (<i>ChiB</i>) Gene Enhances Resistance against <i>Erysiphae polygoni</i>Induced Powdery Mildew Disease in the Transgenic Black Gram (<i>Vigna mungo</i>L.) (cv. T9)

To enhance the antifungal response of blackgram (Vigna mungo L.), transgenic plants were generated by transferring bacterial chitinase gene with a CaMV 35S promoter. The chopped multiple shoot cells developed on the cotyledonary node were transformed by Particle gun method. Thecalli were raised on the Murashige and Skoog (MS) modified media supplemented with 50 m·gl-1 kanamycin. The transformation efficiency was 13% approximately. The resultant shoot buds were selected and the antibiotic resistant transgenic plantlets were regenerated. The development of the transgenic plants from the shoot buds took about four to six months. The integration of the transgene was confirmed by PCR, RT-PCR, Southern and western blot analyses. The transgenic plants exhibited higher chitinase activity than the non-transformed ones. The chitinase activity was examined by native polyacrylamide in-gel assay. The transgenic plants showed fungal tolerance as evidenced by the delayed onset of the disease and smaller lesions following an in vitro inoculation of the powdery mildew pathogen (Erysiphae polygoni DC). The transgenic plants adapted well to the greenhouse and did not show any phenotypic alterations.

Chronic toxicity of both raw and processed Polygoni Multiflori Radix on rats

Recently, adverse effects of Polygoni Multiflori Radix（PMR） and Polygoni Multiflori Radix Praeparata（PMRP） have attracted intensive attention worldwide. These adverse effects most occurred in cases with high dose of prolonged medication course. Liver is usually the target organ of these adverse effects. In the present research, we performed in vivo chronic toxicity study and aimed to evaluate relationships between major constituents of water extractions and total anthraquinone of PMR and PMRP and chronic toxicity. SD rats of both sexes were given water extractions as well as total anthraquinone of PMR and PMRP for 12 weeks. We evaluated basic biochemical indexes, conducted microscopic observations of main organs and assessed early indicators of liver and renal fibrosis. Simvastatin, with hypertriglyceridemia and hypercholesterolemia as its main therapeutic areas, was investigated in our study. Component-toxicity relationships were also discussed. Five male rats died in our study, while all female rats survived, suggesting that some gender differences might be involved. Body weight was significantly changed, and basic biochemical indexes were sporadically occurred during the research. Pathological examinations on liver and kidney showed slight alternations after 12 weeks without dose-dependent relationship. Increase in serum laminin（LN） was observed in almost all male rats, indicating that the risks of liver or kidney fibrosis still existed, especially for males, although no fibrosis was found in the pathological examination of liver and kidney. No major and severe adverse effects were observed after 12 weeks of administration of PMR and PMRP. Regular safety monitoring is still necessary during medication in order to prevent possible risks.

Enhanced absorption and inhibited metabolism of emodin by 2,3,5,4＇-tetrahydroxystilbene-2-O-β-D-glucopyranoside： Possible mechanisms for Polygoni Multiflori Radix-induced liver injury

Polygoni Multiflori Radix(PMR) has been commonly used as a tonic in China for centuries. However, PMR-associated hepatotoxicity is becoming a safety issue. In our previous in vivo study, an interaction between stilbenes and anthraquinones has been discovered and a hypothesis is proposed that the interaction between stilbene glucoside-enriching fraction and emodin may contribute to the side effects of PMR. To further support our previous in vivo results in rats, the present in vitro study was designed to evaluate the effects of 2, 3, 5, 4'-tetrahydroxystilbene-2-O-β-D-glucopyranoside(TSG) on the cellular absorption and human liver microsome metabolism of emodin. The obtained results indicated that the absorption of emodin in Caco-2 cells was enhanced and the metabolism of emodin in human liver microsomes was inhibited after TSG treatment. The effects of the transport inhibitors on the cellular emodin accumulation were also examined. Western blot assay suggested that the depressed metabolism of emodin could be attributed to the down-regulation of UDP-glucuronosyltransferases(UGTs) 1A8, 1A10, and 2B7. These findings definitively demonstrated the existence of interaction between TSG and emodin, which provide a basis for a better understanding of the underlying mechanism for PMR-induced liver injury.

Chemical profiles and metabolite study of raw and processed Polygoni Multiflori Radix in rats by UPLC-LTQ-Orbitrap MS^n spectrometry

The raw and processed roots of Plygonum multiflorum Thunb(PM) are used to treat different diseases in clinical practice. In order to clarify the influence of processing, a comparative study of chemical substance analysis was carried out. As the xenobiotics with a high enough exposure in target organs being considered as the potential effective or toxicity components, an in vivo study was also implemented to characterize the constitutes and metabolites, and meanwhile, the factor of compatibility with black bean were also considered. As a result, a total of 148 compounds were detected in PM extracts and more than 40 compounds were only detected in the processed products, which were probably new components produced during the steaming process. In in vivo study, 7 prototype components and 66 metabolites were detected or tentatively identified, 24 of which were reported for the first time. Our results indicated that processing greatly changed the chemical composition of PM and influenced the disposition of the compounds in vivo. To the best of our knowledge, this was the first global comparative study of raw and processed PM. These results expanded our knowledge about the influence of processing of PM and provided the essential data for further efficacy or toxicity studies.

A high expression EGFR/cell membrane chromatography and online high performance liquid chromatography/mass spectrometry method for screening EGFR antagonists from Rhizoma Polygoni Cuspidati

The epidermal growth factor receptors(EGFRs)in some tumor cells are significant targets for drug discovery.In this work,we have developed an EGFR cell membrane chromatography and online high performance liquid chromatography/mass spectrometry system for screening active component from Rhizoma Polygoni Cuspidati.As a result,resveratrol from Rhizoma Polygoni Cuspidati was found to be the active component acting on EGFR like gefitinib.There was a good relationship between their inhibiting effects on EGFR secretion and HEK293 EGFR cell growth in vitro.The EGFR/CMC-online-HPLC/MS system demonstrated fast and effective characteristics for screening leading compounds from traditional Chinese medicine.

Hematopoietic effect of small molecular fraction of Polygoni multiflori Radix Praeparata in cyclophosphamide-induced anemia mice

The aim of this study is to investigate the protective effects of a small molecular fraction(SMF) of Polygoni multiflori Radix Praeparata(PMRP) in a cyclophosphamide(CTX) induced anemia mouse model. Small molecular fraction of PMRP was prepared and identified by high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry(HPLC-Q-TOF-MS). In pharmacology, we examined the peripheral hemogram and thymus and spleen index. The content of granulocyte-macrophage colony-stimulating factor(GM-CSF) in serum was mensurated by enzyme-linked immunosorbent assay(ELISA);The level of superoxide dismutase(SOD), catalase(CAT), total antioxidant capacity(T-AOC), and malondialdehyde(MDA) in serum and spleen tissue homogenate were detected, and glutathione peroxidase(GSH-PX) was assayed in spleen. The results show that SMF can significantly accelerate the recovery of peripheral hemogram, increase the activity of antioxidant enzymes and GM-CSF in serum and spleen. SMF also increases the number of spleen cells, improves bone marrow pathology. In conclusion, the SMF of PMRP promoted the recovery of hematopoietic function in a CTX-induced anemia mouse, which can support SMF to be used as an adjunct to chemotherapy to counteract its side effects.

Effect of Polygoni Cuspidati Rhizoma et Radix and Its Ingredient Resveratrol on Experimental Autoimmune Myasthenia Gravis by Suppressing Immune Response

Objective To investigate the effects of Polygoni Cuspidati Rhizoma et Radix （PCRR） and its ingredient resveratrol （Res） on experimental autoimmune myasthenia gravis （EAMG）. Methods EAMG was induced in Lewis rats by the immunization of a synthetic peptide corresponding to region 97-116 of the rat acetylcholine receptor （AChR） α subunit （R97-116）. EAMG rats were randomly divided into PCRR group, Res group, and control （C） group, and were ig administered respectively with PCRR （2 g/kg）, Res （20 mg/kg）, and DMSO （0.4 mL/kg） every day from day 5 after immunization to day 42. Clinical evaluation, lymphocyte proliferation, cytokines, and anti-97-116 antibodies were performed for examination of their therapeutic effects. Results Treatments with PCRR and Res significantly ameliorated clinical symptoms, down-regulated TNF-α and up-regulated IL-10 in serum and culture supernatants of lymphocytes stimulated with R97-116, and decreased levels of anti-R97-116 IgG1 and IgG2a in serum compared with C group. Unexpectedly, PCRR but not Res inhibited lymphocyte proliferation compared with C group. Conclusion PCRR and Res ameliorating EAMG is associated with suppressing immune response, and indicates a therapeutic potential for EAMG and even human myasthenia gravis （MG）. Res may be the main effective ingredient from PCRR ameliorating EAMG, but further experiments are necessary.

Effect of Rhizoma Polygoni Cuspidati and Ramulus Cinnamomi Compatibility on Uric Acid Metabolism and Urinary Neutrophil Gelatinase-Associated Lipocalin and Kidney Injury Molecule-1 in Rats with Hyperuricemia

Objective：To explore the effects of Rhizoma Polygoni Cuspidati and Ramulus Cinnamomicompatibility（PR） on uric acid metabolism and the expression of urinary neutrophil gelatinase-associated lipocalin（NGAL） and kidney injury molecule-1（KIM-1） in rats with hyperuricemia. Methods：Seventy male Sprague Dawley（SD） rats were randomly divided into 7 groups with 10 rats per group, including the normal group, model group, allopurinol group, benzbromarone group and PR groups at 3 doses（3.5, 7, 14 g/kg）. Except the normal group, rats of the other groups were intragastrically administered 100 mg/kg hypoxanthine and 250 mg/kg ethambutol, and subcutaneously injected with 200 mg/kg potassium oxonate. All rats were continuously modeled for 17 days, and gavaged with corresponding drugs. The rats of the normal and model groups were gavaged with saline, once a day, for 2 weeks. The levels of serum uric acid（SUA）, blood urea nitrogen（BUN） and creatinine（Cr） were determined. In addition, the contents of NGAL and KIM-1 in urine and the m RNA and protein expressions of xanthine oxidase（XOD） in liver of hyperuricemia rats were measured by reverse transcription polymerase chain reaction（RT-PCR） and Western blot, respectively. Moreover, the pathological changes of kidney were analyzed by hematoxylin and eosin（HE） stain method. Results：Compared with the normal group, the levels of SUA, BUN, NGAL and KIM-1 and the expressions of hepatic XOD m RNA and protein in the hyperuricemia rats were increased significantly（P〈0.01）. PR significantly decreased the levels of SUA, BUN, NGAL and KIM-1 and down-regulated the m RNA and protein expressions of hepatic XOD（P〈0.05 or P〈0.01）. In addition, the pathological changes of kidney were significantly suppressed by oral administration of PR. Conclusions：PR ameliorated uric acid metabolism and protected renal function, the underlying mechanism was mediated by decreasing the levels of SUA, BUN, NGAL and KIM-1, inhibiting the expression of hepatic XOD and ameliorating the pathological change of kidney.

Meridian Tropism of Polygoni Avicularis Herba by Tissue Distribution of Avicularin in Rats

Objective To observe the meridian tropism of Polygoni Avicularis Herba by tissue distribution of avicularin in rats. Methods Tissue distribution of avicularin in rat following a single iv administration was appointed and observed. HPLC method was established and validated for the determination of avicularin in rat tissues. Results Kidney and bladder were the most important target tissue of avicularin. Conclusion HPLC method is successfully applied to tissues distribution study of avicularin after iv administration to rats, and the results explain Polygoni Avicularis Herba on bladder tropism reasonably.

Efficacy of Heshouwu(Radix Polygoni Multiflori)on gut mircobiota in mice with autoimmune encephalomyelitis

OBJECTIVE:To learn the mechanisms between gut microbiome and the autoimmunity benefits on Traditional Chinese Medicine(TCM)in central nervous system(CNS),we investigated the neuro-protection effects and gut mircobiota changes of Heshouwu(Radix Polygoni Multiflori)on experimental autoimmune encephalomyelitis(EAE),an animal model of multiple sclerosis(MS).METHODS:Mice were randomly divided into four groups:EAE mice(control phosphate-buffered saline group),50 mg·kg^(-1)·d^(-1)Heshouwu(Radix Polygoni Multiflori)-treated EAE mice,100 mg·kg^(-1)·d^(-1)Heshouwu(Radix Polygoni Multiflori)-treated EAE mice,and 200 mg·kg^(-1)·d^(-1)Heshouwu(Radix Polygoni Multiflori)-treated EAE mice.The spinal cords were stained with hematoxylin and eosin(HE)and luxol fast blue for evaluating inflammatory infiltration and demyelination.The percentages of granulocyte macrophage-colony stimulating factor(GM-CSF)+CD4+,interleukin 17(IL-17)+CD4+,Foxp3 CD4+,and interferon-γ(IFN-γ)+CD4+T cells in the inguinal lymph nodes(LNs)and brain were determined by flow cytometry analysis.16S rRNA gene sequencing was employed to analyze the changes in gut microbiota.RESULTS:We found that Heshouwu(Radix Polygoni Multiflori)alleviated the disease severity and neuropathology of EAE as evaluated by clinical and histopathologyical scores.Heshouwu(Radix Polygoni Multiflori)increased the diversity and abundance of the gut microbiota,and decreased Firmicutes/Bacteroidetes ratio(F/B ratio).Heshouwu(Radix Polygoni Multiflori)also decreased the concentrations of IL-10,and IL-21 and increase the levels of GM-CSF,IL-17A,IL-17F and IL-22 in serum of EAE mice.Moreover,Heshouwu(Radix Polygoni Multiflori)modulated the T cell responses by inhibiting Th17 cells and restoring Treg cells in the small intestine lymphoid tissues and inguinal lymph nodes.Microbiota-depleted mice receiving Heshouwu(Radix Polygoni Multiflori)-treated fecal microbiota transplantation had lower disease severity,neuropathology scores and alleviation of Th17/Treg imbalance compared to ad libitum group.CONCLUSIONS:Our findings suggested that the vital neuro-protection role of Heshouwu(Radix Polygoni Multiflori)(TCM)in immunomodulation effects partly by regulations of gut microbiome.

Spectrum-Toxicity Correlation Study Revealed the Influence of the Nine-Time Steaming and Sun Drying Method on Hepatotoxic Components of Polygoni Multiflori Radix

Objective:Polygoni Multiflori Radix(PM)is a traditional herbal medicine with repeated reports of liver injury events in recent years.We wondered whether the classical processing method,namely,nine-time steaming and sun drying(NSSD),had toxicity-attenuating effects on PM and the relationships between toxicity and times of processing,as well as with the alteration trends of its compounds.Materials and Methods:The chemical fingerprints of different PM extracts were developed using ultra-high-performance liquid chromatography.The spectrum-toxicity correlation between the chemical fingerprints and hepatocellular toxicity was analyzed with multiple correlation analysis.Results:The results suggested that the hepatotoxicity of NSSD processing products markedly decreased with the repeated steaming and sun drying,which was obviously superior to the product processed by the modern method.Comprehensive analysis revealed that the contents of cis-stilbene glycoside and emodin-8-O-β-D-glucoside related to liver injury susceptibility were reduced with the times of NSSD processing,which was consistent with the decreased trend of hepatocellular toxicity.After the five times of NSSD,the contents of them as well as the hepatotoxicity of PM were steady.Moreover,we found that the contents of catechin and physcion declined rapidly after the one time of NSSD and then remained stable until the nine times of NSSD.Based on the fact,they could be utilized to indicate whether PM products were processed by steaming and sun drying.Conclusions:This paper confirmed that the NSSD had a good influence on the toxicity attenuating to PM and found four compounds which could apply for the quality control of PM.

Screening and Identifying Hepatotoxic Components in Polygoni multiflori Radix and Polygoni multiflori Radix Praeparata

Objective: In this study, the hepatotoxic components of Polygoni multiflori Radix and Polygoni multiflori Radix Praeparata(known as Heshouwu [HSW] and Zhiheshouwu [ZHSW] in China, respectively) were screened, isolated, and identified. Materials and Methods: The ethanol extracts of HSW and ZHSW were separated into 80 fractions according to their polarity in the preparation liquid phase. Chang liver cell line was used to screen the toxic components of HSW and ZHSW in vitro. The obtained toxic mixture was further collected, isolated, and identified to confirm the hepatotoxic compounds of HSW and ZHSW. Results: The identifid hepatotoxic compounds include 2,3,5,4’-tetrah ydroxystilbene-2-O-β-D-glucoside, emodin, physcion-8-O-β-d-glucoside, physcion, and citreorosein, the fist two among them were the main components of HSW and ZHSW. After processing of HSW, the contents of 2,3,5,4’-tetrahydroxystilbene-2-O-β-D-glucoside and emodin in ZHSW were signifiantly decreased. Conclusions: The traditional processing with herb has signifiant effects on the components, especially the toxic components, in the extract of HSW and is an effective method to reduce its toxicity.

Simultaneous Determination of Four Anthraquinones in Polygoni Multiflori Radix with Single Reference Standard by High Performance Liquid Chromatography

Objective: To establish a rapid, accurate and reliable analytical method for the simultaneous determination of four major anthraquinones in Polygoni Multiflori Radix(PMR) using single reference standard.Methods: The four components including emodin-8-O-β-D-(EMG), physcion-8-O-β-D-glucoside, emodin and physcion were separated on an ODS C18 column within 13 min and detected at 280 nm. Emodin was selected as the reference standard, and the response factor for each analyte with respect to emodin were calculated. Robustness were also tested including different columns, equipments, temperatures, detection wavelengths, and other chromatographic conditions which might influence stability of response factors.Results: The method was validated in terms of linearity(r^2> 0.9995), LOQs(0.820–3.05 ng/m L), LODs(0.180–0.920 ng/m L), precision,accuracy(95.8–103.6%, RSD < 2.80%) and stability. A total of 40 batches of PMR were analyzed and the results were found to have no statistically significant differences compared with those obtained using the external standard method.Conclusion: This work provided a single standard to determine multi-components method for quantitation of four anthraquinones in PMR,which could be applied in the quality control of this herbal drug.