Efficacy of Heshouwu () 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 () on experimental autoimmune encepha-lomyelitis (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·d Heshouwu ()-treated EAE mice, 100 mg·kg·d Heshouwu ()-treated EAE mice, and 200 mg·kg·d Heshouwu ()-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 () alleviated the disease severity and neuropathology of EAE as evaluated by clinical and histopathologyical scores. Heshouwu () increased the diversity and abundance of the gut microbiota, and decreased / ratio (F/B ratio). Heshouwu () 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 () 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 ()-treated fecal microbiota trans-plantation 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 () (TCM) in immunomodulation effects partly by regulations of gut microbiome.

Polygoni Multiflori Radix interferes with bile acid metabolism homeostasis by inhibiting Fxr transcription, leading to cholestasis.

Objective: To explore the possible mechanisms of cholestasis induced by Polygoni Multiflori Radix (PM). Methods: Low and high doses of water extract of PM were given to mice by gavage for 8 weeks. The serum biochemical indexes of aspartate aminotransferase (AST), alanine aminotransferase (ALT), glutamyltransferase (GGT) alkaline phosphatase (ALP) and so on were detected in the second, fourth, sixth, and eighth weeks after administration. At the end of the eighth week of administration, the bile acid metabolic profiles of liver and bile were screened by high-performance liquid chromatography tandem triple quadrupole mass spectrometry (HPLC-QQQ-MS/MS). Liver pathological changes were observed by hematoxylin and eosin staining. Real-time quantitative polymerase chain reaction (RT-qPCR) was used to detect the mRNA transcription of the target genes and Western blotting (WB) was used to the detect target protein expression. Results: Biochemical tests results showed the values of ALP and GGT were two and three times greater than the normal values respectively, and the value of R was less than 2. Histopathology also showed that PM caused lymphocyte infiltration, a small amount of hepatocyte necrosis and nuclear fragmentation in mouse liver. The proliferation of bile duct epithelial cells was observed in the high group. These results indicated that PM may lead to cholestatic liver injury. HPLC-QQQ-MS/MS analysis with the multivariate statistical analysis revealed significant alterations of individual bile acids in liver and gallbladder as compared to those of the control group. RT-qPCR showed that the transcription of Fxr, Shp, Bsep, Bacs, Mdr2, and Ugt1a1 were downregulated and that of Cyp7a1, Mrp3, and Cyp3a11 was significantly upregulated in the treatment group. WB demonstrated that PM also markedly downregulated the protein expression of FXR, BSEP, and MDR2, and upregulated CYP7A1. Conclusion: PM inhibited the expression of FXR, which reduced the expression of MDR2 and BSEP, leading to the obstruction of bile acids outflow, and increased the expression of CYP7A1, resulting in an increase of intrahepatic bile acid synthesis, which can lead to cholestasis.

Comparative Investigation of Raw and Processed Radix Polygoni Multiflori on the Treatment of Vascular Dementia by Liquid Chromatograph-Mass Spectrometry Based Metabolomic Approach.

Radix Polygoni Multiflori (PM) is a well-known nootropic used in traditional Chinese medicine (TCM). Considering the efficacy and application discrepancy between raw (RPM) and processed PM (PPM), the similarities and differences between them in the treatment of vascular dementia (VaD) is intriguing. In this study, a VaD rat model was constructed by 2-vessel occlusion (2-VO). During 28 days of treatment, plasma was collected on days 7, 14, 21, and 28 after the start of dosing and the metabolic profile was analyzed by HPLC-MS/MS-based metabolomics. The Morris Water Maze Test, hematoxylin-eosin and Nissl staining, and biochemical analysis were used to assess cognitive function, pathogenic alterations and oxidative stress, respectively. RPM and PPM effectivelyreducedthe 2VO-induced cognitive impairment and mitigated histological alterations in hippocampus tissue. The 2-VO model significantly elevated MDA level and decreased SOD activity and GSH level, indicating severe oxidative stress, which could also be attenuated by RPM and PPM treatment. RPM outperformed PPM in decreasing MDA levels while PPM outperformed RPM in increasing GSH levels. Differential metabolites were subjected to Metabolite Set Enrichment Analysis (MSEA) and genes corresponding to proteins having interactions with metabolites were further annotated with Gene Ontology (GO). Both RPM and PPM ameliorated VaD-relevant vitamin B6 metabolism, pentose phosphate pathways, and taurine and hypotaurine metabolism. In addition, the metabolism of cysteine and methionine was regulated only by RPM, and riboflavin metabolism was modulated only by PPM. The results suggested that raw and processed PM had comparable efficacy in the treatment of VaD but also with some mechanistic differenece.

Pharmacokinetics, Tissue Distribution, and Excretion Characteristics of a Radix Polygoni Multiflori Extract in Rats.

Although progress has been achieved in the pharmacological activity and toxicity of Radix Polygoni Multiflori (RPM), the chemical basis of its toxicity is still unclear. Here, we performed a multicompound pharmacokinetic analysis and investigated the tissue distribution and excretion characteristics of RPM components after oral administration in rats. The findings demonstrated that the active ingredients of the RPM extract were quickly absorbed after oral administration, with high exposure levels of emodin, 2,3,5,4'-teterahydroxystilbene-2-O-ß-D-glucoside (TSG), citreorosein, torachrysone-8-O-glucoside (TG), emodin-8-O-ß-D-glucoside (EG), and physcion-8-O-ß-D-glucoside (PG). The tissue distributions of emodin, TSG, TG, EG, and PG were high in the liver and kidney. These components were the key contributors to the effectiveness and toxicity of RPM on the liver and kidney. Most of the active ingredients were mainly excreted through feces and bile, while a few were converted into other products in the body and excreted through urine and feces.

Overview of Pharmacokinetics and Liver Toxicities of Radix Polygoni Multiflori.

Radix Polygoni Multiflori (RPM), a traditional Chinese medicine, has been used as a tonic and an anti-aging remedy for centuries. However, its safe and effective application in clinical practice could be hindered by its liver injury potential and lack of investigations on its hepatotoxicity mechanism. Our current review aims to provide a comprehensive overview and a critical assessment of the absorption, distribution, metabolism, excretion of RPM, and their relationships with its induced liver injury. Based on the well-reported intrinsic liver toxicity of emodin, one of the major components in RPM, it is concluded that its plasma and liver concentrations could attribute to RPM induced liver injury via metabolic enzymes alteration, hepatocyte apoptosis, bile acids homeostasis disruption, and inflammatory damage. Co-administered 2,3,5,4'-tetrahydroxystilbene-2-O-ß-D-glucopyranoside in RPM and other drugs/herbs could further aggravate the hepatotoxicity of emodin via enhancing its absorption and inhibiting its metabolism. To ensure the safe clinical use of RPM, a better understanding of the toxicokinetics and effect of its co-occurring components or other co-administered drugs/herbs on the pharmacokinetics of emodin is warranted.

A NMR-based drug screening strategy for discovering active substances from herbal medicines: Using Radix Polygoni Multiflori as example.

ETHNOPHARMACOLOGICAL RELEVANCE: Herbal medicines have always been important sources for new drugs. And developing new drugs from traditional herbal medicine is currently still an effective way. However, screening for active substances from herbal medicines extracts has ever been a challenging topic, due to their intrinsic complexity. The herb Radix Polygoni Multiflori has been used as a tonic and an antiaging herb in Traditional Chinese Medicine. In clinical studies, the extract of Radix Polygoni Multiflori can improve hypercholesterolemia, atherosclerotic, diabetes and other diseases commonly associated with glycolipid metabolism, however, the molecular mechanisms of these actions are unknown. AIM OF THE STUDY: We devised a NMR-based drug screening strategy for discovering active substances from herbal medicines, using Radix Polygoni Multiflori as example to address such challenging topic, meanwhile, to explore molecular target of Radix Polygoni Multiflori's glycolipid metabolism benefit. MATERIALS AND METHODS: Herbal medicines extracts were subjected to moderate separation to generate libraries of pre-purified subfractions, target protein was then added to each subfraction, and ligand-observed NMR experiments (line-broadening experiment, chemical shift perturbations measurements and saturation transfer difference spectrum) were performed, active substances identification and structural optimization were then accomplished using signals provided by ligand-observed NMR interaction detection and HPLC-SPE-NMR. The strategy was demonstrated by discovering an active component from extract of herb Radix Polygoni Multiflori, using human fatty acid binding protein 4 (FABP4) as target protein. RESULTS: 2,4-dihydroxy-6-[(1E)-2-(4-hydroxyphenyl)ethenyl]phenyl-ß-D-glucopyranoside(TSG), the hit from one subfraction, has obvious interaction with target protein FABP4, due to FABP4 is a potential therapeutic target for metabolic diseases such as diabetes and atherosclerosis, the screening result will give clue to the active component and molecular target of Radix Polygoni Multiflori's glycolipid metabolism benefit. Besides, interaction information at atom level offered by ligand-observed NMR experiment would be valuable in the further stage of lead optimization. CONCLUSIONS: The devised NMR-based drug screening strategy can discover active substances from herbal medicines efficiently and precisely, meanwhile, can shed light on molecular mechanism of traditional usage of the herb.

Quantification of eight active ingredients in crude and processed radix polygoni multiflori applying miniaturized matrix solid-phase dispersion microextraction followed by UHPLC.

A rapid, efficient, and green sample preparation method has been developed to extract eight active ingredients (gallic acid, catechins, epicatechin, polydatin, 2,3,5,4'-tetrahydroxystilbene-2-O-ß-d-glucoside, resveratrol, emodin, and physcion) in radix polygoni multiflori by miniaturized matrix solid-phase dispersion microextraction. Simple and sensitive ultra high performance liquid chromatography combined with ultraviolet detection has been applied to analyze the multiple compounds. The best results were obtained by adding 25 mg sample into 25 mg adsorbent and grinding for 2 min with disorganized silica as adsorbent and 1 mL 150 mM 1-dodecyl-3-methylimidazolium bromide as a green eluting solvent. Good linearity (r2  > 0.998) for each analyte was obtained by this method. The intra-day and inter-day precision (RSD) were both below 5.31%, and the recoveries of the analytes ranged from 93.3 to 100.0%. This simple miniaturized matrix solid-phase dispersion microextraction method for analyzing the compounds in radix polygoni multiflori needs a short time and requires little sample and reagent. Thus, this method is far more eco-friendly and efficient than traditional extraction methods (reflux and ultrasound-assisted extraction). The present investigation provided a promising method for the fast preparation and discrimination of chemical differences in crude and processed radix polygoni multiflori.

Discrimination of Radix Polygoni Multiflori from different geographical areas by UPLC-QTOF/MS combined with chemometrics.

BACKGROUND: Nowadays, Radix Polygoni Multiflori (RPM, Heshouwu in Chinese) from different geographical origins were used in clinic. In order to characterize the chemical profiles of different geographical origins of RPM samples, ultra-high performance liquid chromatography quadrupole time of flight mass spectrometry (UPLC-QTOF/MS) combined with chemometrics (partial least squared discriminant analysis, PLS­DA) method was applied in the present study. METHODS: The chromatography, chemical composition and MS information of RPM samples from 18 geographical origins were acquired and profiled by UPLC-QTOF/MS. The chemical markers contributing the differentiation of RPM samples were observed and characterized by supervised PLS­DA method of chemometrics. RESULTS: The chemical composition differences of RPM samples derived from 18 different geographical origins were observed. Nine chemical markers were tentatively identified which could be used as specific chemical markers for the differentiation of geographical RPM samples. CONCLUSIONS: UPLC-QTOF/MS method coupled with chemometrics analysis has potential to be used for discriminating different geographical TCMs. Results will help to develop strategies for conservation and utilization of RPM samples.

A Network-Based Pharmacology Study of the Herb-Induced Liver Injury Potential of Traditional Hepatoprotective Chinese Herbal Medicines.

Herbal medicines are widely used for treating liver diseases and generally regarded as safe due to their extensive use in Traditional Chinese Medicine practice for thousands of years. However, in recent years, there have been increased concerns regarding the long-term risk of Herb-Induced Liver Injury (HILI) in patients with liver dysfunction. Herein, two representative Chinese herbal medicines: one-Xiao-Chai-Hu-Tang (XCHT)-a composite formula, and the other-Radix Polygoni Multiflori (Heshouwu)-a single herb, were analyzed by network pharmacology study. Based on the network pharmacology framework, we exploited the potential HILI effects of XCHT and Heshouwu by predicting the molecular mechanisms of HILI and identified the potential hepatotoxic ingredients in XCHT and Heshouwu. According to our network results, kaempferol and thymol in XCHT and rhein in Heshouwu exhibit the largest number of liver injury target connections, whereby CASP3, PPARG and MCL1 may be potential liver injury targets for these herbal medicines. This network pharmacology assay might serve as a useful tool to explore the underlying molecular mechanism of HILI. Based on the theoretical predictions, further experimental verification should be performed to validate the accuracy of the predicted interactions between herbal ingredients and protein targets in the future.

Study on idiosyncratic liver injury and content of cis-2,3,5,4'-tetrahy­droxystilbene-2-O-β-D-glucoside in radix Polygoni multiflori Preparata / 药学学报

This study was designed to investigate the correlation between idiosyncratic liver injury and content of cis-2,3,5,4'-tetrahydroxystilbene-2-O-β-D-glucoside(cis-SG)in radix Polygoni multiflori Preparata(RPMP). In order to compare the effect of hepatotoxicity of different cis-SG contents in RPMP, rats were administered with 50% alcohol extracts of RPMP(7.56 g·kg-1, via intragastric administration)alone or co-treated with lipopolysaccharide(LPS, 2.8 mg·kg-1, via tail vein injection). The results showed that no significant alterations of plasma ALT and AST activities were observed in the normal rats. In the LPS treated rats, the group without light treatment and the group with 0.10% cis-SG after light treatment did not exhibit obvious injury in liver. The group with 0.35% cis-SG after light treatment and the group with 0.70% cis-SG after light treatment showed significant increases in ALT, AST, TNF-α, IL-6, NF-κB p65 and apoptosis rate(P < 0.05), causing pathological changes in the liver tissue. Through the content analysis of drug in patients with liver injury, we found that the content of cis-SG(> 0.40%)was generally higher than that of pieces collected from different origins(< 0.10%). The comparative analysis of experiments and clinical data showed that there was a relationship between the content of cis-SG and idiosyncratic liver injury. In order to reduce the risk of clinical medication, the content of cis-SG of 0.10% should be a limit of quality control in the production processing of Polygonum multiflorum.

Quality evaluation of Heshouwu, a Taoist medicine in Wudang, China.

Polygonum multiflorum Thunb., which is known as Heshouwu in China, is a Taoist medicine sourced from the Wudang mountain area. At present, the quality of the Heshouwu sourced from this region is unknown. The present study aimed to evaluate the quality of wild Heshouwu collected from the Wudang mountain area, particularly the 2,3,5,4'-tetrahydroxystilbene-2-O-ß-D-glucoside (TSG) and combined anthraquinone (CAQ) content, compared with that of commercially available Heshouwu. Furthermore, the potential quantities of organic pesticide residues were determined. High performance liquid chromatography with a diode array detector was used to quantify TSG and CAQ content, whereas gas chromatography (GC), performed using a temperature gradient, was used to detect the presence of organochlorine, pyrethroid and organophosphorus pesticides. The average TSG content present in the wild Heshouwu from the Wudang mountain area and in the commercially available Heshouwu was 2.39 and 1.10%, respectively. In addition, the average content of CAQ in these was 1.41 and 3.46%, respectively. GC did not detect residues of organic pesticides in the wild Heshouwu, thus this plant met the criterion of the Chinese Pharmacopeia (2010 edition). The results of the present study indicated that wild Heshouwu from the Wudang mountain area may be suitable for use as a Chinese medicine across China.

Novel functional polysaccharides from Radix Polygoni Multiflori water extracted residue: Preliminary characterization and immunomodulatory activity.

The alkali-extractable polysaccharides (APMPs) were isolated from the water extracted residues of Radix Polygoni Multiflori, and further purified by DEAE-52 cellulose and Sephadex G-100 column chromatography to obtain a homogeneous polysaccharide (APMP-2) with molecular weights of 7724.8 Da. HPLC chromatography analysis identified that APMP-2 was a heteropolysaccharides and mainly composed of Galactose and Xylose with a molar ratio of 4.31: 1.06. It was shown that both APMP and APMP-2 were of activation effects on splenocytes and peritoneal macrophages, and also significantly restore the proliferation rate, phagocytic index and cytokine (IL-2 and TNF-α) production level of 5-FU-treated splenocytes/peritoneal macrophages in a dosage-dependent manner. The results suggested that polysaccharides presented in Radix Polygoni Multiflori water-extracted residues possessed immunomodulatory activity and could be used as potential immunomodulators, and this finding could be a reference for the utilization of Radix Polygoni Multiflori water extracted residues.

Characterization of the constituents in rat plasma after oral administration of radix polygoni multiflori extracts by ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometry.

An ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometry method was established to detect as many constituents in rat plasma as possible after oral administration of Radix polygoni multiflori (RPM) extract. A C18 column (150 × 2.0 mm, 4 µm) was adopted to separate the samples, and mass spectra were acquired in negative modes. The fingerprints of RPM extract were established, resulting in 39 components being detected. Among these compounds, 29 were identified by comparing the retention times and mass spectral data with those of reference standards and relevant references, and eight compounds were separated and detected in RPM for the first time. In vivo, 23 compounds were observed in dosed rat plasma, 16 of 23 compounds were indicated as prototype components of RPM, and seven compounds were predicted to be metabolites of RPM. A high-speed and sensitive method was developed and was successfully utilized for screening and characterizing the ingredients and metabolites of RPM.

Effects of stilbene glucoside on lipid synthesis, decomposition and transport in steatotic L-02 hepatocytes / 中国药学杂志

OBJECTIVE: To elucidate the mechanism of the active chemical substance of Radix Polygoni Multiflori which is an important Yunyao with lipid-lowering activity, 2, 3, 5, 4'-tetrahydroxystilbene-2-O-β-D-glucoside (TSG), for regulating lipid metabolism, thus making a foundation for further studies of Radix Polygoni Multiflori.

Effective components, toxic effects and research advances of Radix Polygoni Multiflori / 国际药学研究杂志

With the wide use of Radix Polygoni Multiflori and it's preparations in the field of medicine, cosmetology, food and healthcare, the adverse reactions generated subsequently have also received considerable attention gradually. In this article, the main effective components of Radix Polygoni Multiflori and their pharmacological effects, toxicity and the possible mechanism, adverse reactions in clinical applications are reviewed.

The in vitro and in vitro antioxidant activities of the ethyl acetate extracted from Radix Polygoni multiflori

Radix Polygoni multiflori has been used Vietnamese traditional medicine for a long time. This paper presented the remarkable antioxidant effects of the AcOET extract from Radix Polygoni multiflori with evidences obtained from various antioxidant tests: inhibition of lipid peroxidation in cell hemogenates, of anion superoxide production, stimulation of SOD activity using Ransod kits. The extract protected the splenocytes from H20¬2- induced injuries and protected brain cells from oxidative stress caused by an ischemia/reperfusion course. These results suggested that application of Radix Polygoni multiflori as antioxidant agent will be beneficial in prevention and treatment of diseases of the elderly

Study on Quality Standard of Jingwu Oral Liquid / 中国药房

OBJECTIVE:To improve the quality standard for Jingwu oral liquid.METHODS:The qualitation of Radix Polygoni Multiflori preparata and solomonseal rhizome in Jingwu oral liquid were identified by TLC and the assaying of 2,3,5,4’—tetra—hydroxyl diphenyl ethylene—2—O—?—D—glucoside in the oral liquid were determined by HPLC.RESULTS:In the TLCs of Radix Polygoni Multiflori preparata and solomonseal rhizome,the color of speckles was the same as that of the control articles in the corresponding places.The linear range of 2,3,5,4’—tetra—hydroxyl diphenyl ethylene—2—O—?—D—glucoside was 0.0 562?g～0.6 744?g(r=0.9 996);The average recovery was 100.1%(RSD=1.89%).CONCLUSION:The method is simple,fast,and reproducible and of high recovery rate,which can be used as quality control for Jingwu oral liquid.

Effect of Extracts of Herba(RPMP) on the Growth and Proliferation of Cultured Melanocytes / 中国医师杂志

Objective To study of the effect of traditional Chinese medicine(TCM): Herba radix polygoni multiflori preparata (RPMP) on the cultured melanocytes(MC). Methods Extracts of TMC RPMP treated the cultured melanocytes, and tyrosinase activity, melanin level and proliferation of the cells were determined. Meanwhile, method of micropore filter was used to measure the ability of cell migration. Results TCM RPMP markedly promoted the proliferation and migration of MC in a dose-dependent manner at a range of 0～150?g/ml. Conclusions TCM RPMP can promote the proliferation and migration of MC in vitro. And it suggests that this drug could be used to treat vitiligo.

Effect of Protein and Anthraquinone Glycosides from Radix Polygoni Multiflori on Learning and Memory and Metabolites in D-galactose-induced Aging Mice / 广州中医药大学学报

Objective To observe the effects of protein and anthraquinones glycosides from Radix Polygoni Multiflori(RPM) on learning and memory as well as associated enzymes and metabolites in mice with D-galactose-induced aging.Methods Eighty Kunming mice were equally randomized into 8 groups: normal group,model group,vitamin E(90mg?kg-1?d-1) group,high-and low-dose RPM protein(20 and5 mg?kg-1?d-1 respectively) groups,high-and low-dose RPM anthraquinone glycoside(28 and 7mg?kg-1?d-1 respectively) groups,combination(low-dose RPM protein and low-dose RPM anthraquinone glycosides)group.Except the normal group,the mice in other groups received subcutaneous injection of D-galactose to indue the aging model.The learning rate of mice was detected with trisection radiation maze,memory was evaluated by one-time passive avoidance response,cerebral superoxide dismustase(SOD) and monoamine oxidase(MAO) activities as well as malondialdehyde(MDA)content were detected with ultraviolet spectrophotometer,and hepatic lipofuscin(LF) content was examined with fluorescence spectrophotometer.Results Memory of aged mice was much improved in high-dose RPM protein group(P

Studies on nootropic activity and mechanism of emodin-8-O-?-D-glucopyranoside / 中草药

Object To investigate the nootropic activity of emodin 8 O ? D glucopyranoside, the active principle of Radix Polygoni Multiflori Preparata(PMEG), and its mechanism of action Methods Nootropic activity was evaluated with step down test of mice, and its nootropic mechanism studied by its antiacetylcholinesterase (anti AchE) effect on mice brain in vivo and in vitro as determined by pectrophotometry Results PMEG (ig) significantly improved the learning and memory of both normal and intelectual deficit mice induced by scopolamine PMEG showed dose dependent inhibition of AchE, and the antiacetylcholinesterase effect is reversible both in vivo and in vitro In acute experiment, the half recovery time of AchE is 115 min, and 165 min in chronic experiment Conclusion PMEG showed good nootropic activity, and it is a reversible AchE inhibitor