Polygonum Multiflorum Extract Research Articles

Alleviation of Glucocorticoid-Induced Osteoporosis in Rats by Ethanolic Reynoutria multiflora (Thunb.) Moldenke Extract.

Secondary osteoporosis is frequently due to the use of high-dose glucocorticoids (GCs). The existing strategy for managing glucocorticoid-induced osteoporosis (GIOP) is considered insufficient and remains in a state of ongoing evolution. Therefore, it is crucial to develop more precise and effective agents for the treatment of GIOP. The constituents of Reynoutria multiflora (Thunb.) Moldenke, specifically Polygonum multiflorum (PM) Thunb, have previously shown promise in mitigating osteopenia. This study aimed to investigate the therapeutic effects of an ethanolic PM extract (PMR30) against GIOP in male rats. Prednisone (6 mg/kg/day, GC) was continuously administered to rats to induce GIOP, and they were subjected to treatment with or without ethanolic PMR30 for a duration of 120 days. Serum was collected for biochemical marker analysis. Bone histomorphometric, histological, and TUNEL analyses were performed on tibia samples. The protein expressions of LC3, Agt5, and Beclin 1 in the femur underwent examination through western blotting. Prolonged and excessive GC treatment significantly impeded bone formation, concomitant with reduced bone mass and body weight. It also suppressed OCN and OPG/RANKL in serum, and decreased Beclin 1 and LC3 in bone. Simultaneously, there was an elevation in bone resorption markers and apoptosis. Treatments with both high dose and low dose of PMR30 alleviated GIOP, stimulated bone formation, and upregulated OCN and OPG/RANKL, while suppressing TRACP-5b, CTX-I, and apoptosis. The impact of PMR30 possibly involves the enhancement of autophagy proteins (LC3, Agt5, and Beclin 1) and the inhibition of apoptosis within the bone. PMR30 holds promise as a prospective therapeutic agent for preventing and treating GIOP.

Induction of Nrf2-EpRE-mediated gene expression by hydroxyanthraquinones present in extracts from traditional Chinese medicine and herbs.

Hydroxyanthraquinones that can be present in traditional Chinese medicine (TCM) and herbal extracts have claimed beneficial intestinal effects. We examined the ability of a panel hydroxyanthraquinones, and methanolic extracts from selected TCM and herbal granules to activate Nrf2-EpRE mediated gene expression using a reporter-gene assay. The results indicate that purpurin, aloe-emodin, 2-hydroxy-3-methylanthraquinone and rhein induced Nrf2 mediated gene expressions with a high induction factor (IFs>10), with BMCL10 values (the lower confidence limit of the concentration giving 10% added response above background) of 16 μM, 1.1 μM, 23 μM and 2.3 μM, respectively, while aurantio-obtusin, obtusifolin, rubiadin 1-methyl ether and emodin were less potent (IFs<5), with BMCL10 values for added response above background level of 4.6 μM, 15 μM, 9.8 μM and 3.8 μM, respectively. All TCM extracts and the herbal extracts of Aloe Vera, Polygonum multiflorum, Rubia (cordifolia) and Rheum officinale activated the Nrf2-EpRE pathway. Of the TCM extracts, Chuan-Xin-Lian-Kang-Yan-Pian was the most potent Nrf2-inducer. LC-MS/MS analysis indicated the presence of selected hydroxyanthraquinones in the extracts and herbs, in part explaining their Nrf2-EpRE mediated activity. In conclusion, different hydroxyanthraquinones have different potencies of Nrf2 activation. The Nrf2 activation by extracts from TCM and herbs can be partially explained by the presence of selected hydroxyanthraquinones.

Composite double-layer microneedle loaded with traditional Chinese medicine for the treatment of androgenic alopecia

Androgenetic alopecia (AGA) is an androgen-mediated alopecia affected by both genes and hormones. Medication is a relatively common treatment. As a new drug delivery method, microneedles (MNs) can effectively break through the stratum corneum barrier, deliver drugs more efficiently, and achieve better therapeutic effects. In this study, we develop a composite double-layer MN through multi-step casting fabrication using a polydimethylsiloxane mold. The needle tip was fabricated by mixed solution of chitosan and polyvinylpyrrolidone which was loaded with Polygonum multiflorum extract, and the base layer was prepared by mixed solution of polyvinyl alcohol and polyvinylpyrrolidone. In vitro mechanical tests showed that the maximum load of a single tip of the drug-loaded MN was about 3.5 ​N, which met the mechanical requirements of skin puncture (>1 ​N). The drug release experiment showed that the MN could achieve gradual drug release. In the animal experiment, pigmentation and hair regrowth occurred earlier in the Polygonum multiflorum-MN (Pm-MN) group than in the other groups, and hair growth finally appeared in almost the entire area. Compared with the AGA model mice, mice in the Pm-MN group achieved an increase in the number and diameter of hair follicles. In conclusion, the Pm-MN is scientific and feasible for treating AGA.

A Network-Pharmacology-Combined Integrated Pharmacokinetic Strategy to Investigate the Mechanism of Potential Liver Injury due to Polygonum multiflorum

Polygonum multiflorum (PM) has been used as a tonic and anti-aging remedy for centuries in Asian countries. However, its application in the clinic has been hindered by its potential to cause liver injury and the lack of investigations into this mechanism. Here, we established a strategy using a network pharmacological technique combined with integrated pharmacokinetics to provide an applicable approach for addressing this issue. A fast and sensitive HPLC-QQQ-MS method was developed for the simultaneous quantification of five effective compounds (trans-2,3,5,4'-tetrahydroxystilbene-2-O-β-d-glucoside, emodin-8-O-β-d-glucoside, physcion-8-O-β-d-glucoside, aloe-emodin and emodin). The method was fully validated in terms of specificity, linearity, accuracy, precision, extraction recovery, matrix effects, and stability. The lower limits of quantification were 0.125-0.500 ng/mL. This well-validated method was successfully applied to an integrated pharmacokinetic study of PM extract in rats. The network pharmacological technique was used to evaluate the potential liver injury due to the five absorbed components. Through pathway enrichment analysis, it was found that potential liver injury is primarily associated with PI3K-Akt, MAPK, Rap1, and Ras signaling pathways. In brief, the combined strategy might be valuable in revealing the mechanism of potential liver injury due to PM.

Correlation between the redox activity of Polygonum multiflorum extract and its extraction technology with Chinese liquor (Baijiu): An electrochemistry-based study

Elucidating the pharmaceutical mechanisms behind traditional Chinese medicine (TCM) is the key to promote its modernization process. In China, soaking TCM in liquor has a history of thousands of years, and many TCMs have to be processed into liquor before they can be used to treat diseases. Chinese liquor (Baijiu) contains more than 2,000 trace components, the interaction mechanism between TCM and Baijiu still remains unclear, making TCM a "mystery". The TCM industry commonly employs chromatographic and spectrographic technology to investigate the redox activity of TCM substances. However, only investigating the redox differences in specific active substances cannot provide a complete understanding of the redox activity of TCM substances. Thus, we employed the electrochemical approach to study the overall redox activity of substances in TCM in situ. The key result is that the redox substances in Baijiu function as a mediator for the redox reaction of Polygonum multiflorum extract. The redox efficiency of the extract is enhanced because of the faster electron transferability of the redox mediator in Baijiu.

Hypoglycemic effects of naturally processed Polygonum multiflorum extract in KK CgAy/J mice and its mechanism of action

In 2021, there are approximately 537 million adults ageing 20 79 years affected by diabetes worldwide and the number is rising rapidly, hence it is important to manage and control diabetes mellitus and its associated complications. Food is one of the key factors in preventing and combating diseases such as diabetes. Both as a food and an herbal medicine, Polygonum multiflorum (PM) has been used as an anti-aging tonic, for hair darkening in traditional Chinese medicine for several centuries. The recent research effort of PM has been focused on antioxidant, anti-ageing and anti-tumor properties. In the present study, we utilized the traditional processing of harvested raw PM, and identified several stilbene components and then evaluated the potential anti-diabetic effects of the processed PM extract (PME). PME (0.075 %) was given to diabetic mice (KK CgAy/J) in drinking water and after 7 weeks, PME-treated mice had significantly lower glucose levels than mice in the diabetic control group ( P < 0.01). The mechanism was explored with ELISA and Western blotting and results suggested that the effect was through maintaining β-cell function.

Polygonum multiflorum extract support hair growth by elongating anagen phase and abrogating the effect of androgen in cultured human dermal papilla cells

BackgroundDermal papilla cells (DPCs) play a key role in hair growth among the various cell types in hair follicles. Especially, DPCs determine the fate of hair follicle such as anagen to telogen transition and play a pivotal role in androgenic alopecia (AGA). This study was performed to elucidate the hair growth promoting effects of Polygonum multiflorum extract (PM extract) in cultured human DPCs and its underlying mechanisms.MethodsThe effects of PM extract on cultured DPCs were investigated. Cell viability and mitochondrial activity were measured by CCK-8 and JC-1 analysis, respectively. Western blotting, dot blotting, ELISA analysis, immunocytochemistry and real-time PCR analysis were also performed to elucidate the changes in protein and mRNA levels induced by PM extract. 3D cultured DPC spheroids were constructed for mimicking the in vivo DPs. The hair growth stimulatory effect of PM extract was evaluated using human hair follicle organ culture model.ResultsPM extract increased the viability and mitochondrial activity in cultured human DPCs in a dose dependent manner. The expression of Bcl2, an anti-apoptotic protein expressed dominantly in anagen was significantly increased and that of BAD, a pro-apoptotic protein expressed in early catagen was decreased by PM extract in cultured DPCs and/or 3D DPC spheroid culture. PM extract also decreased the expression of catagen inducing protein, Dkk-1. Growth factors including IGFBP2, PDGF and VEGF were increased by PM extract, revealed by dot blot protein analysis. We also have found that PM extract could reverse the androgenic effects of dihydrotestosterone (DHT), the most potent androgen. Finally, PM extract prolonged the anagen of human hair follicles by inhibiting catagen entry in human hair follicle organ culture model.ConclusionOur data strongly suggest that PM extract could promote hair growth by elongating the anagen and/or delaying the catagen induction of hair follicles through activation of DPCs.

The Protective Effects of 2,3,5,4'-Tetrahydroxystilbene-2-O-β-d-Glucoside in the OVA-Induced Asthma Mice Model.

Asthma is an inflammatory disease caused by an imbalance of Th1 and Th2 cells. In general, asthma is characterized by a stronger Th2 response. Most conventional asthma treatment focuses on improving airway flow or suppression of airway inflammation. To reduce the side effects of currently used asthma medicines, we have conducted studies on natural products that have no side effects. 2,3,5,4′-tetrahydroxystilbene-2-O-β-d-glucoside (TSG), the main compound of Polygonum multiflorum (PM), has various biological activities, including anti-inflammation and anti-oxidation activities. However, the effect of TSG on asthma has not been studied yet. We examined the effects of TSG on Th2 immune responses using an OVA-induced asthma animal model. OVA-sensitized mice were treated with TSG. 24 h after the last intranasal challenge, airway hyperresponsiveness (AHR) was measured or serum and bronchoalveolar lavage fluid (BALF) were harvested. We measured typical Th1 and Th2 cytokines in serum and BALF. As a result, TSG suppressed Th2 responses, as shown by the lower levels of IL-4, IL-5, total IgE, OVA-specific IgE, and OVA-specific IgG1. On the other hand, TSG increased Th1 responses, as shown by the levels of IFN-gamma. Collectively, these results confirm the potential of TSG for asthma treatment through modulation of inflammatory responses. Considering that the cytotoxic effect of PM extract is due to the cis isomer of TSG, if the effect of TSG on asthma treatment is found to be non-toxic in clinical trials, it would be more effective to use it as a purified component than PM extract as an asthma treatment agent.

Polygonum multiflorum Extract Exerts Antioxidative Effects and Increases Life Span and Stress Resistance in the Model Organism Caenorhabditis elegans via DAF-16 and SIR-2.1

Extracts of the Chinese plant Polygonum multiflorum (PME) are used for medicinal purposes as well as food supplement due to anti-aging effects. Despite of the common use of these food supplements, experimental data on physiological effects of PME and its underlying molecular mechanisms in vivo are limited. We used the model organism Caenorhabditis elegans to analyze anti-aging-effects of PME in vivo (life span, lipofuscin accumulation, oxidative stress resistance, thermal stress resistance) as well as the molecular signaling pathways involved. The effects of PME were examined in wildtype animals and mutants defective in the sirtuin-homologue SIR-2.1 (VC199) and the FOXO-homologue DAF-16 (CF1038). PME possesses antioxidative effects in vivo and increases oxidative stress resistance of the nematodes. While the accumulation of lipofuscin is only slightly decreased, PME causes a significant elongation (18.6%) of mean life span. DAF-16 is essential for the reduction of thermally induced ROS accumulation, while the resistance against paraquat-induced oxidative stress is dependent on SIR-2.1. For the extension of the life span, both DAF-16 and SIR-2.1 are needed. We demonstrate that PME exerts protective effects in C. elegans via modulation of distinct intracellular pathways.

Studies on constituents of Polygonum multiflorum extract in vivo and in vitro based on UPLC-Q-TOF-MS

To explore the drug-induced constituents in vivo of Polygonum multiflorum extract (PM). This study was the first to study the drug-induced constituents in target organ liver. Agilent MassHunter qualitative analysis software and Metabolite ID software were applied for the analysis of retention time, exact relative molecular mass, primary and secondary mass spectrum information based on ultra performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) and targeted-MS/MS. By comparison with literature and standards, a total of 5 prototypes and 6 metabolites were identified or tentatively elucidated from the liver samples. In addition, the drug-induced constituents in plasma and PM were also analyzed in this study and 8 prototypes and 19 metabolites were detected from the plasma samples, while 30 compounds were detected from the extract of PM. Emodin oxidative acetylation metabolites, hydroxyl methylation metabolites, carboxylation glucuronidation metabolites and ketone glucuronidation metabolites in this study were first reported. Through the comparative analysis between the in vivo and in vitro constituents of PM, the study preliminarily revealed the drug-induced constituents (prototypes and metabolites) in liver and clarified the transfer process and transmutation rules of constituents in PM, blood and liver, which would further deepen our understanding on constituents of PM in vivo.

Interpretation the Hepatotoxicity Based on Pharmacokinetics Investigated Through Oral Administrated Different Extraction Parts of Polygonum multiflorum on Rats.

The liver injury induced by Polygonum multiflorum (PM) used for clinical treatment has recently received widespread attention. This study aimed to determine the hepatotoxicity of PM through pharmacokinetics studies. The extract of PM was separated to isolate the anthraquinone fraction, the tannin and polysaccharide fraction, the hydroxystilbene fraction, and the combined anthraquinone fraction. A rapid LC-MS/MS method was developed and validated to simultaneously analyze 2,3,5,4′-tetrahydroxystilbene-2-O-β-glucoside (TSG), emodin-8-O-β-D-glucopyranoside (EDG), and emodin in rat plasma, and was applied to the pharmacokinetics (PK) studies. The hepatotoxicity of different extracted parts of PM was evaluated through the levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), total bilirubin (TBil), direct bilirubin (DBil), and indirect bilirubin (IBil) in rat serum. The results showed that liver injury occurred in all the treated groups and that the hepatotoxicity performance of the total extract was different from other groups. The pharmacokinetic studies showed that the Cmax, Tmax, AUC, t1/2, and MRT of the major compounds of different extracted parts were significantly different in rat plasma at same dosage. Emodin-O-hex-sulfate, tetrahydroxystilbene-O-(galloyl)-hex, emodin (original and generated through EDG deglycosylation), and other free anthraquinones might be responsible for the hepatotoxicity of PM in vivo. PM extracts produced inhibitory effects on drug metabolic enzymes, include CYP3A4, CYP2C19, CYP2E1, UGT1A1, etc. And these effects may be related to its hepatotoxicity and pharmacokinetic behavior different. This information on hepatotoxicity and the pharmacokinetic comparison may be useful to understand the toxicological effects of PM.

Immunomodulating Properties of Polygonum multiflorum Extracts on Cyclophosphamide-induced Immunosuppression Model

The objective of the present study was to investigate the immunomodulating property of Polygonum multiflorum extracts using various in vitro and in vivo tests. In this study, Polygonum multiflorum extracts were shown to exhibit immunomodulating activities such as cytokine production in normal rat splenocyte and cyclophosphamide-induced immunosuppression model. The immunostimulant effects of Polygonum multiflorum extracts were further tested for various immunological markers. Polygonum multiflorum extracts significantly enhanced cytokines production activity in rat splenocyte. Consistently oral administration of Polygonum multiflorum extracts enhanced blood cytokine production and ameliorated spleen damage in cyclophosphamide immunosuppressed rat. In conclusion, the present investigation revealed that Polygonum multiflorum extracts possessed immunomodulating and immunostimulating properties.

Preventive effects of Polygonum multiflorum on glucocorticoid-induced osteoporosis in rats.

In Traditional Chinese Medicine, Polygonum multiflorum (PM) is known for its anti-aging properties. A previous study by our group showed that extracts of PM were able to prevent and treat bone loss in vivo, and the active components emodin and 2,3,5,4,-tetrahydroxystilbene-2-O-β-glucoside (TSG) promoted the osteogenic differentiation of mesenchymal stem cells in vitro. The aim of the present study was to investigate the preventive effects of PM on glucocorticoid-induced osteoporosis (GIO) in rats. A crude extract of PM was prepared with 75% ethanol, purified and enriched using a D-101 macroresin column and elution with 30% ethanol, and the material obtained was assessed by high-performance liquid chromatography. Male or female Sprague Dawley rats (n=180) were randomly divided into nine groups: Control, prednisone, prednisone plus calcitriol (CAL), prednisone plus 30% ethanolic eluate of PM [high (H), medium (M) and low (L) dose] and prednisone plus crude extract of PM (H, M and L dose). Prednisone was orally administered to the osteoporosis model rats for 21 weeks, alongside which they received PM extracts. The weight of the viscera, anterior tibial muscle and other tissues was recorded at the end of the experiment. The femur and lumbar vertebra were collected for the measurement of three-dimensional microarchitecture by micro-computed tomography scanning, assessment of biomechanical properties and determination of bone mineral density (BMD). In the 30% ethanolic eluate of the PM extract, the content of TSG and combined anthraquinone was 9.20 and 0.15%, respectively, and that in the crude extract of PM was 2.23 and 0.03%, respectively. Over 6 weeks, the weight of the rats the in prednisone group decreased (P<0.05), while the weight of rats treated with M and H doses of 30% ethanolic eluate was increased compared with that in the prednisone group (P<0.05). Rats exposed to prednisone exhibited a deteriorated bone microarchitecture, low BMD, decreased bone volume/total volume and poor biomechanical properties. Furthermore, the weight of the adrenal gland and the anterior tibial muscle was decreased. 30% ethanolic eluate of PM at M and L doses and crude extract of PM at the H dose counteracted the alterations of skeletal and other characteristics induced by prednisone in rats, as did CAL. In conclusion, extracts of PM exerted a protective effect on bone tissue in GIO rats.

Anti-inflammatory effects of trans-2,3,5,4′-tetrahydroxystilbene 2-O-β-glucopyranoside (THSG) from Polygonum multiflorum (PM) and hypoglycemic effect of cis-THSG enriched PM extract

Polygonum multiflorum (PM) has been used as a tonic edible plant and is a popular anti-ageing dietary supplement. Trans-2,3,5,4′-tetrahydroxystilbene 2-O-β-glucopyranoside (trans-THSG) is the major active compound in PM. It was found to have potent anti-inflammatory effects in RAW 264.7 macrophage cells and in TPA-induced mouse ear edema model. Research in the significance and rational of traditional process of post-harvested PM has been neglected. With the aid of modern analytical technology, we studied the major phytochemical transformation of PM solution from trans- to cis-THSG under UV light, reminiscent of traditional process, and further evaluated the hypoglycemic effects of the processed PM extract. In a high fat diet-induced type 2 diabetic mice model, processed PM extract containing cis- and trans-THSG (ratio=2:3) exhibited significant hypoglycemic capacity, while pure trans-THSG and unprocessed PM extract (cis-/trans-THSG ratio=1:20) did not, suggesting the importance of anti-diabetic effect of cis-THSG and PM natural process.

Application of iTRAQ-Based Quantitative Proteomics Approach to Identify Deregulated Proteins Associated with Liver Toxicity Induced by Polygonum Multiflorum in Rats

Background/Aims: Clinical reports on adverse reactions that result from Polygonum multiflorum (PM) and its preparations, especially regarding liver injury, have recently received widespread attention. This study aimed to investigate the mechanism of hepatotoxicity induced by different PM extracts through iTRAQ quantitative proteomics. Methods: The different PM extracts were orally administrated for 90 days to rats, and the hepatotoxicity effect was evaluated through measurement of biochemical indexes, oxidative damage indexes and hematoxylin-eosin (HE) staining. Then, the hepatotoxicity mechanism was investigated by iTRAQ quantitative proteomics. Results: The results of biochemical and histopathological analyses showed that liver injury occurred in all groups of rats given by various PM extracts, which proved all of the PM extracts could induce hepatotoxicity. The hepatotoxicity mechanism may differ between the total extract group and the other groups through the results of biochemical indicators. The iTRAQ proteomics study showed that hepatotoxicity resulting from PM was mainly related to the abnormal activity of mitochondrion function-related oxidative phosphorylation pathways. Conclusion: This iTRAQ proteomics study revealed that the hepatotoxicity induced by PM is primarily related to the oxidative phosphorylation pathways. NADH dehydrogenase family proteins and Slc16a2 could be potential biomarkers of hepatotoxicity resulting from PM.

Neuroprotective effects of herbal cocktail on cerebrovascular dysfunction in rats with induced hyperhomocysteinaemia

Introduction: Hyperhomocysteinaemia (HHcy) is an established risk factor for cardiovascular, cerebrovascular, peripheral vascular diseases and neurodegenerative disease. The effect of this HHcy on vascular diseases could potentially cause vascular pathology features. Experimental studies have demonstrated that Hcy can be neurotoxic to brain, hippocampus area. &#x0D; Methods: The present study was conducted to compare the possible neuroprotective effects of different herbal cocktail in HHcy-induced rats’ brain cerebrovascular dysfunction model. Rats were divided into nine groups: Group I - Controls received the same volume of saline solution (0.5 mL/100 g of body weight). Group II served as HHcy and received homocysteine 0.03 μmol/g of b.w. daily for 30 days. Group III served as HHcy and received homocysteine 0.03 μmol/g of b.w. + Artemisia Judaica extract (AJ) (50 mg/kg per oral by oral feeding needle with tuberculin syringe) daily for 30 days. Group IV served as HHcy and received homocysteine 0.03 μmol/g of b.w.+ Panax ginseng extract (PG) (50 mg/kg per oral by oral feeding needle with tuberculin syringe) daily for 30 days. Group V served as HHcy and received homocysteine 0.03 μmol/g of b.w. + Polygonum multiflorum extract (PM) (400 mg/kg per oral by oral feeding needle with tuberculin syringe) daily for 30 days. Group VI served as HHcy and received homocysteine 0.03 μmol/g of b.w. + AJ + PG with the same dose of previous group daily for 30 days. Group VII served as HHcy and received homocysteine 0.03 μmol/g of b.w. + AJ + PM with the same dose of the previous group daily for 30 days. Group VIII served as HHcy and received homocysteine 0.03 μmol/g of b.w. + PG + PM with the same dose of the previous group daily for 30 days. Group IX served as HHcy and received homocysteine 0.03 μmol/g of b.w. + AJ + PG + PM with the same dose of previous group daily for 30 days. The hippocampus of brain samples was collected at the end of the experiment and measuring oxidative stress markers (CAT, SOD, MDA and NO), inflammatory mediators (IL-6 and BDNF), histopathological examination and comet assay.&#x0D; Results: Revealed data showed that the homocysteine induces SOD, CAT depletion, and an increase in AChE, MDA, NO, IL-6, and BDNF. A mixture of PG and PM or the individual treatments showed an ameliorative response for all parameters. In general, oxidative stress parameters, inflammatory mediator, neurotrophic factor, pathological examination and comet were degenerate against HHcy but did not differ significantly compared to AJ group.&#x0D; Conclusion: Better physiological and histological characteristics were in PG and PM and their combination groups compared with HHcy and ameliorated nearly the control group.

Metabolomic Study on Idiosyncratic Liver Injury Induced by Different Extracts of Polygonum multiflorum in Rats Integrated with Pattern Recognition and Enriched Pathways Analysis.

Currently, numerous liver injury cases related to a famous Chinese herb- Polygonum Multiflorum (Heshouwu in Chinese) have attracted great attention in many countries. Our previous work showed that Heshouwu-induced hepatotoxicity belonged to idiosyncratic drug-induced liver injury (IDILI). Unfortunately, the components and mechanisms attributed to IDILI of Heshouwu are difficult to determine and thus remain unknown. Attempts to explore puzzles, we prepared the chloroform (CH)-, ethyl acetate (EA)-, and residue (RE) extracts of Heshouwu to investigate IDILI constituents and underlying mechanisms, using biochemistry, histopathology, and metabolomics examinations. The results showed that co-treatment with non-toxic dose of lipopolysaccharide (LPS) and EA extract could result in evident liver injury, indicated by the significant elevation of plasma alanine aminotransferase and aspartate aminotransferase activities, as well as obvious liver histologic damage; whereas other two separated fractions, CH and RE extracts, failed to induce observable liver injury. Furthermore, 21 potential metabolomic biomarkers that differentially expressed in LPS/EA group compared with other groups without liver injury were identified by untargeted metabolomics, mainly involved two pathways: tricarboxylic acid cycle and sphingolipid metabolism. This work illustrated EA extract had close association with the idiosyncratic hepatotoxicity of Heshouwu and provided a metabolomic insight into IDILI of different extracts from Heshouwu.

Novel compound from Polygonum multiflorum inhibits inflammatory response in LPS-stimulated microglia by upregulating AMPK/Nrf2 pathways

Polygonum multiflorum extracts are known to improve memory and learning ability, and have neuroprotective and anti-aging activity. However, its function and the underlying mechanisms in neuroinflammation-mediated neurodegenerative disease remain poorly understood. In the present study, we investigated the anti-neuroinflammatory effects of several compounds from P. multiflorum, and found a novel compound, CRPE55IB. The CRPE55IB-induced suppression of NO and PGE2 production correlated with inhibition of iNOS and COX-2 protein expression and promoter activity in lipopolysaccharide (LPS)-stimulated microglia. CRPE55IB also reduced the production of pro-inflammatory cytokines (TNF-α and IL-6) induced by LPS. Furthermore, investigation of the molecular mechanism indicated that CRPE55IB inhibited LPS-induced NF-κB activation by inactivating phosphorylation of IKKα/β, and phosphorylation and degradation of IκBα. We further found that CRPE55IB inhibited the phosphorylation of ERK and JNK at a lower concentration than that for p38 MAPK. Further experiments revealed that CRPE55IB treatment considerably increased the activation of Nrf2/ARE, and the expression of its target genes, including HO-1 and NQO1. Moreover, the Knockdown of Nrf2, HO-1, and NQO1 by siRNA abrogated the inhibitory effect of CRPE55IB on iNOS and COX-2 promoter activity. CRPE55IB also induced phosphorylation of AMPK/LKB/CaMKII in microglia. Analysis using a specific inhibitor of AMPK demonstrated that AMPK activation was involved in CRPE55IB-induced HO-1 and NQO1 expression. In addition, the CRPE55IB-induced anti-neuroinflammatory effect was abrogated by a specific inhibitor of AMPK, indicating the important role of AMPK in CRPE55IB-induced anti-neuroinflammation. Collectively, these results demonstrate that CRPE55IB exerts anti-neuroinflammatory effects against LPS via the Nrf2/AMPK signaling pathways.

Influence of specification on chemical composition of dissolution and hepatocytes toxicity of Polygonum multiflorum

According to different toxicities of various aqueous extracts of Polygonum multiflorum on hepatocyte, the impacts of chemical composition on the safety of P. multiforum was studied. In this study, 8 main chemical compositions in aqueous extracts of P. multiflorum were determined by the established HPLC method; at the same time, the inhibition ratios of different aqueous extracts of P. multiflorum on L02 cell were determined. Afterwards, the potential compounds related to the toxicity of P. multiforum were tentatively found through a multiple correlation analysis. The results showed that P. multiforum with different chemical compositions exhibited great differences in dissolution. The hepatocyte toxicity of P. multiflorum powder was much greater than P. multiflorum lumps. In addition, three constituents closely related to toxicity of P. multiflorum were found by multiple correlation analysis. The study revealed that chemical composition of P. multiflorum is closely related to the hepatotoxicity, and the hepatotoxicity of P. multiflorum powder is greater than that of other dosage forms. This study indicates that P. multiflorum with different chemical compositions show varying toxicity, which therefore shall be given high attention.

Polygonum multiflorum extracts protect the SH-SY5Y cells against the oxidative stress injury induced by MPP+

Objective: To investigate the protective effect of Polygonum multiflorum extracts (EPM) on SH-SY5Y cells treated with MPP+. Methods: SH-SY5Y cells were first exposed to various doses of EPM, and then treated with MPP+. The cell viability was detected with MTT assay, cell morphology was observed with microscope by Hoechst33258 staining, the level of glutathione (GSH), Malondialdehyde (MDA) and the activity of Lactate dehydrogenase (LDH) were measured with UV spectrophotometer. Reactive oxygen species (ROS) and the mitochondrial membrane potential were observed with fluorescence microscope. The expression of p-JNK and Caspase 3 was analyzed by Western Blot. Results: After 0.5 mmol/L MPP+ treatment for 48 h, the viability of SH-SY5Y cells was decreased to 44.7% (VS. control group), the shrunk cell body and nuclear condensation were observed. Compared with control group, the activity of lactate dehydrogenase (LDH), the level of malondialdehyde (MDA) and reactive oxygen species (ROS) and the expression of p-JNK and Caspase 3 were increased significantly in MPP+ treated SH-SY5Y cells, whereas the level of GSH and the mitochondrial membrane potential was reduced significantly in MPP+ treated group. However, pretreatment with EPM (at the concentration of 5, 25 and 100 mg/L) 4 h prior to being exposed to MPP+ rescued the cell viability of SH-SY5Y cells, and restored the cell morphological features in a dose-dependent manner. The increasement of lactate dehydrogenase (LDH), malondialdehyde (MDA) and reactive oxygen species (ROS) and the expression of p-JNK and Caspase 3 induced by MPP+ were also inhibited significantly by pretreatment with EPM. Exposure to EPM also blocked the reduction of the GSH level and the mitochondrial membrane potential induced by MPP+ in SH-SY5Y cells. Conclusion: Our results suggested that the EPM was able to protect the SH-SY5Y cells against the damage induced by MPP+, with the mechanism involved in resisting the oxidative stress.