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A study on the phytochemical composition of the roots of Polygonum multiflorum collecting in Vietnam and an evaluation of its capacity to inhibit pancreatic lipase enzyme was conducted. In total, 12 metabolites (1 – 12) were isolated from anti-lipase activity-guided fractionation of P. multiflorum. The isolated compounds (1 – 12) were identified as trans -resveratrol (1), methyl protocatechuate (2), methyl gallate (3), catechin (4), epicatechin (5), myrciaphenone A (6), quercetin (7), kaempferol (8), apigenin (9), luteolin (10), tricin (11), and afzelin (12). This is the first time Compound 12 has been reported from P. multiflorum. However, 12 second metabolites (1 – 12) were tested in vitro to assess the anti-lipase potential. The results revealed Compound 7 exhibited the most significant lipase inhibition, with an IC50 value of 0.30 ± 0.04 μM. Following closely, Compounds 6, 8, 10, and 12 displayed IC50 values of 5.38 ± 0.65, 0.72 ± 0.10, 0.79 ± 0.19, and 1.22 ± 0.18 μM, respectively. For comparison, orlistat (positive control), the most common anti-obesity drug available on the market, has an IC50 value of 0.91 ± 0.10 μM. Molecular docking was applied to explore the affinity and interactions between active compounds 6 – 8, 10, and 12 and proteins associated with obesity, namely pancreatic lipase–colipase complex (PLCC) and pancreatic lipase-related protein 1 (PLRP1). In addition, absorption, distribution, metabolism, excretion, and toxicity (ADMET) were used in predictions. The results indicate that the active compounds isolated from P. multiflorum could be further in-depth research on lipase inhibition. [ABSTRACT FROM AUTHOR]

This review aims to gain insight into the major causes of hair graying (canities) and how plant-derived extracts and phytochemicals could alleviate this symptom. Research articles on human hair graying were searched and selected using the PubMed, Web of Science, and Google Scholar databases. We first examined the intrinsic and extrinsic factors associated with hair graying, such as the reduced capacity of melanin synthesis and transfer, exhaustion of melanocyte stem cells (MSCs) and melanocytes, genetics and epigenetics, race, gender, family history, aging, oxidative stress, stress hormones, systematic disorders, nutrition, smoking, alcohol consumption, lifestyle, medications, and environmental factors. We also examined various plants and phytochemicals that have shown a potential to interfere with the onset or progression of human hair graying at different levels from in vitro studies to clinical studies: the extract of Polygonum multiflorum and its major components, 2,3,5,4′-tetrahydroxystilbene-2-O-β-D-glucoside and emodin; the extract of Eriodictyon angustifolium and its major flavonoid compounds, hydroxygenkwanin, sterubin, and luteolin; the extracts of Adzuki beans (Vigna angularis), Fuzhuan brick tea (Camellia sinensis), and Gynostemma pentaphyllum; bixin, a carotenoid compound found in Bixa orellana; and rhynchophylline, an alkaloid compound found in certain Uncaria species. Experimental evidence supports the notion that certain plant extracts and phytochemicals could alleviate hair graying by enhancing MSC maintenance or melanocyte function, reducing oxidative stress due to physiological and environmental influences, and managing the secretion and action of stress hormones to an appropriate level. It is suggested that hair graying may be reversible through the following tactical approaches: selective targeting of the p38 mitogen-activated protein kinase (MAPK)–microphthalmia-associated transcription factor (MITF) axis, nuclear factor erythroid 2-related factor 2 (NRF2), or the norepinephrine–β2 adrenergic receptor (β2AR)–protein kinase A (PKA) signaling pathway. [ABSTRACT FROM AUTHOR]

The aim of the experiment was to study the effects of the alcohol extract of Polygonum multiflo rum Thunb. fermented by yeast on the liver and immune cells of mice. Sixty male Kunming mice were randomly divided into four groups. The low-dose group, medium-dose group and high-dose group were respectively injected with a fermented Heshouwu alcohol solution equivalent to the raw drug dose of 11, 21, 42 g/kg, and the control group was treated with double distilled water for 30 days. The results showed that there was no obvious toxicity in the naked eye of the mice during the administration period, the body weight increased steadily, and there was no statistically significant difference in liver coefficient, spleen coefficient, and kidney coefficient. Serum biochemical indexes showed that the levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) in the high-dose group were significantly higher than those in the other groups (P<0.05). ELISA results showed that the level of tumor necrosis factor-α (TNF-α) in the high-dose group was significantly higher than that in the other groups (P<0.05). IL-17, Bax and Bcl-2 were not statistically different (P>0.05). H.E. staining showed a small amount of inflammatory infiltration in the high-dose group. The results of flow cytometry showed that there was no statistical difference in the percentage of immune cells T cells (CD3+, CD4+, CD8+ ) and B cells (CD19+ ) between the spleen of each group of mice (P>0.05). These results indicated that the alcohol extract of yeast-fermented polygonum multiflorum showed no obvious toxicity to mice at low and medium doses. The yeastfermented Polygonum multiflorum had no effect on the immune cells in the spleen of mice. [ABSTRACT FROM AUTHOR]

Objectives: Cognitive impairments, ranging from mild to severe, adversely affect daily functioning, quality of life, and work capacity. Despite significant efforts in the past decade, more than 200 promising drug candidates have failed in clinical trials. Herbal remedies are gaining interest as potential treatments for dementia due to their long history and safety, making them valuable for drug development. This review aimed to examine the mechanisms behind the effect of Polygonum multiflorum on cognitive function. Methods: This study focused primarily on the effects of Polygonum multiflorum and its chemical constituents on cognitive behavioral outcomes including the Morris water maze, the passive avoidance test, and the Y maze, as well as pathogenic targets of cognitive impairment and Alzheimer’s disease (AD) like amyloid deposition, amyloid precursor protein, tau hyperphosphorylation, and cognitive decline. Additionally, a thorough evaluation of the mechanisms behind Polygonum multiflorum’s impact on cognitive function was conducted. We reviewed the most recent data from preclinical research done on experimental models, particularly looking at Polygonum multiflorum’s effects on cognitive decline and AD. Results: According to recent research, Poligonum multiflorum and its bioactive components, stilbene, and emodin, influence cognitive behavioral results and regulate the pathological target of cognitive impairment and AD. Their mechanisms of action include reducing oxidative and mitochondrial damage, regulating neuroinflammation, halting apoptosis, and promoting increased neurogenesis and synaptogenesis. Conclusion: This review serves as a comprehensive compilation of current experiments on AD and other cognitive impairment models related to the therapeutic effects of Polygonum multiflorum. We believe that these findings can serve as a basis for future clinical trials and have potential applications in the treatment of human neurological disorders.

Synucleinopathies, typified by Parkinson’s disease (PD), entail the accumulation of α-synuclein (αSyn) aggregates in nerve cells. Various αSyn mutants, including the αSyn A53T variant linked to early-onset PD, increase the propensity for αSyn aggregate formation. In addition to disrupting protein homeostasis and inducing proteostatic stress, the aggregation of αSyn in PD is associated with an imbalance in iron metabolism, which increases the generation of reactive oxygen species and causes oxidative stress. This study explored the impact of αSyn A53T expression in transgenic hairy roots of four medicinal plants (Lobelia cardinalis, Artemisia annua, Salvia miltiorrhiza, and Polygonum multiflorum). In all tested plants, αSyn A53T expression triggered proteotoxic stress and perturbed iron homeostasis, mirroring the molecular profile observed in human and animal nerve cells. In addition to the common eukaryotic defense mechanisms against proteostatic and oxidative stresses, a plant stress response generally includes the biosynthesis of a diverse set of protective secondary metabolites. Therefore, the hairy root cultures expressing αSyn A53T offer a platform for identifying secondary metabolites that can ameliorate the effects of αSyn, thereby aiding in the development of possible PD treatments and/or treatments of synucleinopathies.

Synucleinopathies, typified by Parkinson's disease (PD), entail the accumulation of α-synuclein (αSyn) aggregates in nerve cells. Various αSyn mutants, including the αSyn A53T variant linked to early-onset PD, increase the propensity for αSyn aggregate formation. In addition to disrupting protein homeostasis and inducing proteostatic stress, the aggregation of αSyn in PD is associated with an imbalance in iron metabolism, which increases the generation of reactive oxygen species and causes oxidative stress. This study explored the impact of αSyn A53T expression in transgenic hairy roots of four medicinal plants (Lobelia cardinalis, Artemisia annua, Salvia miltiorrhiza, and Polygonum multiflorum). In all tested plants, αSyn A53T expression triggered proteotoxic stress and perturbed iron homeostasis, mirroring the molecular profile observed in human and animal nerve cells. In addition to the common eukaryotic defense mechanisms against proteostatic and oxidative stresses, a plant stress response generally includes the biosynthesis of a diverse set of protective secondary metabolites. Therefore, the hairy root cultures expressing αSyn A53T offer a platform for identifying secondary metabolites that can ameliorate the effects of αSyn, thereby aiding in the development of possible PD treatments and/or treatments of synucleinopathies. [ABSTRACT FROM AUTHOR]

Abstract Background Polygonum multiflorum (PM), a widely used traditional Chinese medicine herb, is divided into two forms, namely raw polygonum multiflorum (RPM) and polygonum multiflorum praeparata (PMP), according to the processing procedure. Emerging data has revealed the differential hepatotoxicity of RPM and PMP, however, its potential mechanism is still unclear. Methods In our study, we investigated the differential hepatotoxicity of RPM and PMP exerted in C57BL/6 mice. First, sera were collected for biochemical analysis and HE staining was applied to examine the morphological alternation of the liver. Then we treated L02 cells with 5 mg / mL of RPM or PMP. The CCK8 and EdU assays were utilized to observe the viability and proliferation of L02 cells. RNA sequencing was performed to explore the expression profile of L02 cells. Western blotting was performed to detect the expression level of ferroptosis-related protein. Flow cytometry was used to evaluate ROS accumulation. Results In our study, a significant elevation in serum ALT, AST and TBIL levels was investigated in the RMP group, while no significant differences were observed in the PMP group, compared to that of the CON group. HE staining showed punctate necrosis, inflammatory cell infiltration and structural destruction can be observed in the RPM group, which can be significantly attenuated after processing. In addition, we also found RPM could decrease the viability and proliferation capacity of L02 cells, which can be reversed by ferroptosis inhibitor. RNA sequencing data revealed the adverse effect of PM exerted on the liver is closely associated with ferroptosis. Western blotting assay uncovered the protein level of GPX4, HO-1 and FTL was sharply decreased, while the ROS content was dramatically elevated in L02 cells treated with RPM, which can be partially restored after processing. Conclusions The hepatotoxicity induced by RPM was significantly lower than the PMP, and its potential mechanism is associated with ferroptosis.

Background: Polygonum multiflorum (PM), a widely used traditional Chinese medicine herb, is divided into two forms, namely raw polygonum multiflorum (RPM) and polygonum multiflorum praeparata (PMP), according to the processing procedure. Emerging data has revealed the differential hepatotoxicity of RPM and PMP, however, its potential mechanism is still unclear. Methods: In our study, we investigated the differential hepatotoxicity of RPM and PMP exerted in C57BL/6 mice. First, sera were collected for biochemical analysis and HE staining was applied to examine the morphological alternation of the liver. Then we treated L02 cells with 5 mg / mL of RPM or PMP. The CCK8 and EdU assays were utilized to observe the viability and proliferation of L02 cells. RNA sequencing was performed to explore the expression profile of L02 cells. Western blotting was performed to detect the expression level of ferroptosis-related protein. Flow cytometry was used to evaluate ROS accumulation. Results: In our study, a significant elevation in serum ALT, AST and TBIL levels was investigated in the RMP group, while no significant differences were observed in the PMP group, compared to that of the CON group. HE staining showed punctate necrosis, inflammatory cell infiltration and structural destruction can be observed in the RPM group, which can be significantly attenuated after processing. In addition, we also found RPM could decrease the viability and proliferation capacity of L02 cells, which can be reversed by ferroptosis inhibitor. RNA sequencing data revealed the adverse effect of PM exerted on the liver is closely associated with ferroptosis. Western blotting assay uncovered the protein level of GPX4, HO-1 and FTL was sharply decreased, while the ROS content was dramatically elevated in L02 cells treated with RPM, which can be partially restored after processing. Conclusions: The hepatotoxicity induced by RPM was significantly lower than the PMP, and its potential mechanism is associated with ferroptosis. [ABSTRACT FROM AUTHOR]

The processing of traditional Chinese medicine (TCM) plays an important role in the clinical application, which usually has the function of "increasing efficiency and reducing toxicity". Polygonum multiflorum (PM) has been reported to induce hepatotoxicity, while it is believed that the toxicity is reduced after processing. Studies have shown that the hepatotoxicity of PM is closely related to the changes in chemical components before and after processing. However, there is no comprehensive investigation on the chemical changes of PM during the processing progress. In this research, we established a comprehensive method to profile both small molecule compounds and polysaccharides from raw and different processed PM samples. In detail, an online two-dimensional liquid chromatography coupled with quadrupole-orbitrap mass spectrometry (2D-LC/Q-Orbitrap MS) was utilized to investigate the small molecules, and a total of 150 compounds were characterized successfully. After multivariate statistical analysis, 49 differential compounds between raw and processed products were screened out. Furthermore, an accurate and comprehensive method for quantification of differential compounds in PM samples was established based on ultra-high performance liquid chromatography/Q-Orbitrap-MS (UHPLC/Q-Orbitrap-MS) within 16 min. In addition, the changes of polysaccharides in different PM samples were analyzed, and it was found that the addition of black beans and steaming times would affect the content and composition of polysaccharides in PM significantly. Our work provided a reference basis for revealing the scientific connotation of the processing technology and increasing the quality control and safety of PM. [ABSTRACT FROM AUTHOR]

This review aims to gain insight into the major causes of hair graying (canities) and how plant-derived extracts and phytochemicals could alleviate this symptom. Research articles on human hair graying were searched and selected using the PubMed, Web of Science, and Google Scholar databases. We first examined the intrinsic and extrinsic factors associated with hair graying, such as the reduced capacity of melanin synthesis and transfer, exhaustion of melanocyte stem cells (MSCs) and melanocytes, genetics and epigenetics, race, gender, family history, aging, oxidative stress, stress hormones, systematic disorders, nutrition, smoking, alcohol consumption, lifestyle, medications, and environmental factors. We also examined various plants and phytochemicals that have shown a potential to interfere with the onset or progression of human hair graying at different levels from in vitro studies to clinical studies: the extract of Polygonum multiflorum and its major components, 2,3,5,4′-tetrahydroxystilbene-2-O-β-D-glucoside and emodin; the extract of Eriodictyon angustifolium and its major flavonoid compounds, hydroxygenkwanin, sterubin, and luteolin; the extracts of Adzuki beans (Vigna angularis), Fuzhuan brick tea (Camellia sinensis), and Gynostemma pentaphyllum; bixin, a carotenoid compound found in Bixa orellana; and rhynchophylline, an alkaloid compound found in certain Uncaria species. Experimental evidence supports the notion that certain plant extracts and phytochemicals could alleviate hair graying by enhancing MSC maintenance or melanocyte function, reducing oxidative stress due to physiological and environmental influences, and managing the secretion and action of stress hormones to an appropriate level. It is suggested that hair graying may be reversible through the following tactical approaches: selective targeting of the p38 mitogen-activated protein kinase (MAPK)–microphthalmia-associated transcription factor (MITF) axis, nuclear factor erythroid 2-related factor 2 (NRF2), or the norepinephrine–β2 adrenergic receptor (β2AR)–protein kinase A (PKA) signaling pathway.

ABSTRACT: 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.

Introduction: A growing body of evidence has suggested that Polygonum multiflorum Thunb. (PM) may cause PM-herb-induced liver injury (PM-HILI). However, the compounds triggering PM-HILI remain controversial. Herein, we set out to investigate the relationships among natural products of PM (NPPM), drug-induced liver injury (DILI) positive (POS), and negative (NEG) compounds by using cheminformatics methods. Methods: A total of 197 NPPM and 2384 annotated DILI dataset were collected from the literature. Chemical space, physicochemical properties, drug-likeness, intra-set similarity, and scaffold diversity were compared to gain insights into the multiple features of NPPM. An ensemble machine learning (ML) model was constructed to predict the DILI potential of NPPM. Twelve NPPM were selected and tested on HepaRG cells to validate the prediction results. Results: Results of the principal component analysis suggest that NPPM bears more similarity to NEG in terms of chemical space when compared with POS. Besides, NPPM share a moderate overall scaffold diversity and one-third ingredients in NPPM compiled the drug-like rules. The predictive results of ML model show that 28.9% of the small molecules in NPPM bear DILI potential. Further in vitro study by detecting cytotoxicity of representative compounds on HepaRG cells showed that trans- and cis-emodin-physcion dianthrone exhibited the lowest IC50 values of 53.05 µM and 17.11 µM, respectively. Conclusion: ML methods and further validation recognize the liver toxicity of dianthonres and emodins. A proportion of NPPM components exhibit drug-likeness profiles and might offer complementary resources for drug discovery. The results demonstrated machine learning-powered DILI prediction as a useful tool to study potential DILI risk of compounds, providing a basis for further identification of toxins or leads in PM. The codes used and generated in this study are freely available at https://github.com/dreadlesss/Polygonum_database_analysis.

Objective To explore molecular mechanisms of polygonum multiflorum in the treatment of androgenic alopecia and to provide underlying drug targets. Methods Bioinformatics analysis tool for molecular mechanism of Traditional Chinese Medicine and five disease gene databases were used to establish effective ingredients-targets of polygonum multiflorum network and protein-protein interaction network. The core sub-network was extracted through network topology, and then GO and KEGG analyses were performed. Finally, these top 10 molecules with highest connectivity were performed to molecular docking. Results A total of 1 357 common target genes were generated based on the intersection of effective ingredients-targets of polygonum multiflorum and underlying disease targets. The core sub-network including 67 genes was extracted by topological analysis. The KEGG enrichment results showed that polygonum multiflorum produce biological functions mainly through regulating inflammatory response, apoptosis, oxidative stress and autophagy pathways. The molecular docking results demonstrated the binding energy of PIK3CA-emodin, PIK3CA-chrysophanol, PIK3CA-guaijaverin and PIK3CB-emodin were less -9 kcal/mol. Conclusions The mechanism of polygonum multiflorum to treat androgenic alopecia is that PI3K/Akt pathway was mediated to regulate the expression of PIK3CA and PIK3CB to cell apoptosis through emodin. In addition, it is involved in the process of the inhibition of inflammatory response and oxidative stress, and induction of melanin production.

Ha-Soo-Oh is a traditional Chinese medicine prepared from the roots of Polygonum multiflorum Thunb. The herb extract has been widely used in Asian countries as a tonic agent and nutritional supplement for centuries. To identify new bioactive compounds in Chinese herbs, the biotransformation-guided purification (BGP) process was applied to Ha-Soo-Oh with Bacillus megaterium tyrosinase (BmTYR) as a biocatalyst. The result showed that a major biotransformed compound could be purified using the BGP process with preparative high-performance liquid chromatography (HPLC), and it was confirmed as a new compound, 2,3,5,3′,4′-pentahydroxystilbene-2-O-β-glucoside (PSG) following mass and nucleic magnetic resonance (NMR) spectral analyses. PSG was further confirmed as a biotransformation product from 2,3,5,4′-tetrahydroxystilbene-2-O-β-glucoside (TSG) by BmTYR. The new PSG exhibited 4.7-fold higher 1,1-diphenyl-2-picrylhydrazine (DPPH) free radical scavenging activity than that of TSG. The present study highlights the potential usage of BGP in herbs to discover new bioactive compounds in the future. [ABSTRACT FROM AUTHOR]

Polygonum multiflorum (PM) Thunb., a typical Chinese herbal medicine with different therapeutic effect in raw and processed forms, has been used worldwide for thousands of years. However, hepatotoxicity caused by PM has raised considerable concern in recent decades. The exploration of toxic components in PM has been a great challenge for a long time. In this study, we developed a stepwise strategy integrating metabolomics and pseudotargeted spectrum-effect relationship to illuminate the potential hepatotoxic components in PM. First, 112 components were tentatively identified using ultraperformance liquid chromatography-quadrupole-time-of-flight-mass spectrometry (UPLC-Q-TOF-MS). Second, based on the theory of toxicity attenuation after processing, we combined the UPLC-Q-TOF-MS method and plant metabolomics to screen out the reduced differential components in PM between raw and processed PM. Third, the proposed pseudotargeted MS of 16 differential components was established and applied to 50 batches of PM for quantitative analysis. Fourth, the hepatocytotoxicity of 50 batches of PM was investigated on two hepatocytes, LO2 and HepG2. Last, three mathematical models, gray relational analysis, orthogonal partial least squares analysis, and back propagation artificial neural network, were established to further identify the key variables affecting hepatotoxicity in PM by combining quantitative spectral information with toxicity to hepatocytes of 50 batches of PM. The results suggested that 16 components may have different degrees of hepatotoxicity, which may lead to hepatotoxicity through synergistic effects. Three components (emodin dianthrones, emodin-8-O-ß-D-glucopyranoside, PM 14-17) were screened to have significant hepatotoxicity and could be used as toxicity markers in PM as well as for further studies on the mechanism of toxicity. Above all, the study established an effective strategy to explore the hepatotoxic material basis in PM but also provides reference information for indepth investigations on the hepatotoxicity of PM. [ABSTRACT FROM AUTHOR]

Zheng X, Yang S, Zhang R, Wang S, Li G, Zhou S. Drug Des Devel Ther. 2019;13:3171–3180. The authors have advised due to an error that occurred inadvertently at the time of figure assembly, Figure 5J is incorrect. The correct Figure 5 is shown in Download Article. The authors apologize for this error and advise since this image only displays the cell morphology and does not perform a semi-quantitative analysis, it does not affect the other results of Figure 5, nor does it affect the conclusion of the article. Read the original article

Traditional Chinese Medicines (TCM) are known for their curative effects on hypertension through a holistic approach. The molecular mechanisms of the formulation comprising Polygonum multiflorum, Rehmannia glutinosa, Senna obtusifolia and Crataegus, used by Chinese practitioners in ameliorating hypertension, however remain a mystery. This initial study is thus aimed at unveiling the molecular mechanisms of this TCM formulation in treating hypertension. The methanolic extract compounds of the decoction were identified through Liquid chromatography mass spectrometry-mass spectrometry (LC-MS/MS). Oral bioavailability and drug likeness were then measured to filter out identified compounds. Several databases, such as the SwissTargetPrediction, STRING, OMIM and KEGG, were used to retrieve information on the predicted targets for the purpose of developing a network using Cytoscape Version 3.8. Enrichment analysis was then performed to elucidate the mechanisms of the decoction in hypertension mitigation. A total of 11 compounds identified were revealed to possess bioavailable and drug like characteristics, based on the Veber and Quantitative Estimation of Drug-likeness (QED) parameters. Pathway analysis showed enrichment of pathways such as cardiac muscle contraction, fluid shear stress and atherosclerosis, dilated cardiomyopathy, renin-angiotensin system and hypertrophic cardiomyopathy (HCM), which are all strongly associated with hypertension. The network pharmacology analysis clearly shows that this TCM decoction ameliorates hypertension through several indirect pathways where most of the targets are involved in HCM, which is caused by hypertension.

As a traditional tonic Chinese medicine, Polygonum multiflorum is widely used in clinical practice. However, with the deepening of modern pharmacological research, its drug toxicity, especially hepatotoxicity, has become increasingly prominent. Based on a large number of clinical and experimental evidence, it has been confirmed that Polygonum multiflorum and its main active ingredients such as anthraquinones and diphenylethylene glucoside can cause different degrees of hepatotoxicity. Further studies have shown that the toxicological mechanisms involved in the hepatotoxicity of different extracts and components of Polygonum multiflorum may include oxidative phosphorylation, bile acid excretion, different metabolic pathways, genetic and metabolic factors, immune homeostasis, etc. By sorting out and summarizing the literature related to hepatotoxicity of Polygonum multiflorum in recent years, this paper discussed the hepatotoxicity mechanism of Polygonum multiflorum and its main components and some contradictions in related reports. [Display omitted] • This study discussed the hepatotoxicity mechanism of Polygonum multiflorum and some contradictions in related reports. • The hepatotoxicity mechanism of main components of Polygonum multiflorum was discussed. • Network toxicological study of hepatotoxicity induced by Polygonum Multiflorum was performed. • The hepatotoxicity mechanism may include oxidative phosphorylation, bile acid excretion, different metabolic pathways, etc. [ABSTRACT FROM AUTHOR]

The complexity of chemical components of herbal medicines often causes great barriers to toxicity research. In our previous study, we have found the critical divergent hepatotoxic potential of a pair of stilbene isomers in a famous traditional Chinese herb, Polygonum multiflorum (Heshouwu in Chinese). However, the high-throughput in vitro evaluation for such stereoisomerism-dependent hepatotoxicity is a critical challenge. In this study, we used a hepatic organoids–based in vitro hepatotoxic evaluation system in conjunction with using high content imaging to differentiate in vivo organ hepatotoxicity of the 2,3,5,4′-tetrahydroxy- trans- stilbene-2-O-β-glucoside (trans-SG) and its cis- isomer (cis-SG). By using such an organoid platform, we successfully differentiated the two stereoisomers' hepatotoxic potentials, which were in accordance with their differences in rodents and humans. The lesion mechanism of the toxic isomer (cis-SG) was further found as the mitochondrial injury by high-content imaging, and its hepatotoxicity could be dose-dependently inhibited by the mitochondrial protective agent. These results demonstrated the utility of the organoids-based high-content imaging approach in evaluating and predicting organ toxicity of natural products in a low-cost and high-throughput way. It also suggested the rationale to use long-term cultured organoids as an alternative toxicology platform to identify early and cautiously the hepatotoxic new drug candidates in the preclinical phase. [ABSTRACT FROM AUTHOR]

Can Tu,1,2 Ming Niu,2 Ai-Wu Wei,3 Jin-Fa Tang,3 Le Zhang,2 Jing Jing,2 Xiao-He Xiao,2 Jia-Bo Wang2,4 1Beijing Research Institute of Chinese Medicine, Beijing University of Chinese Medicine, Beijing, 100029, People’s Republic of China; 2China Military Institute of Chinese Medicine, Fifth Medical Center of Chinese PLA General Hospital, Beijing, 100039, People’s Republic of China; 3The First Affiliated Hospital of Henan University of Chinese Medicine, Zhengzhou, 450000, People’s Republic of China; 4School of Traditional Chinese Medicine, Capital Medical University, Beijing, 100069, People’s Republic of ChinaCorrespondence: Jia-Bo WangSchool of Traditional Chinese Medicine, Capital Medical University, Beijing, 100069, People’s Republic of ChinaEmail pharm_sci@126.comXiao-He XiaoChina Military Institute of Chinese Medicine, Fifth Medical Center of Chinese PLA General Hospital, Beijing, 100039, People’s Republic of ChinaEmail pharmacy302xxh@126.comBackground: Drug-induced liver injury is a common adverse effect in clinical practice, with severe cases resulting in liver failure and even death. Identification and prediction of individuals susceptible to idiosyncratic DILI continues to remain a challenge.Methods: In this study, we report that cytokines in human serum can be used to identify and predict individuals susceptible to Polygonum multiflorum-induced DILI (PM-DILI) in retrospective and prospective cohort studies.Findings: In the retrospective pilot study, we compared serum cytokine expression profiles of the PM-DILI group (n=10) and the PM-Tolerant group (n=12) and found 10 cytokines with significant differences. In the replication cohort study, differences in the 10 cytokines between PM-DILI (n =11) and PM-Tolerant (n=13) groups were verified. Among them, 6 cytokines showed no significant differences at two time points, including liver injury and recovery stage of PM-DILI, suggesting that these 6 cytokines have no correlation with PM-DILI, however, they may be related to susceptibility. Furthermore, all the retrospective cohorts were combined, and a PM-DILI susceptibility prediction model was built by screening the 6 cytokines. The combination of (TNF-α and CCL-2) or VEGF showed the highest sensitivity and specificity. Finally, the efficacy of the above 3 cytokine combination models in predicting PM-DILI-susceptible individuals was verified before PM exposure in another independent prospective cohort (n=24), with sensitivity and specificity of 66.7% and 83.3%, respectively.Conclusion: This proof-of-concept study demonstrates that the serum cytokine combination reflecting dysimmunity could be used as a new method to predict PM-DILI, thus providing a new perspective for improving the clinical management of IDILI.Keywords: drug-induced liver injury, Polygonum multiflorum, cytokine, prediction, susceptible individual

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.

The complexity of chemical components of herbal medicines often causes great barriers to toxicity research. In our previous study, we have found the critical divergent hepatotoxic potential of a pair of stilbene isomers in a famous traditional Chinese herb, Polygonum multiflorum (Heshouwu in Chinese). However, the high-throughput in vitro evaluation for such stereoisomerism-dependent hepatotoxicity is a critical challenge. In this study, we used a hepatic organoids–based in vitro hepatotoxic evaluation system in conjunction with using high content imaging to differentiate in vivo organ hepatotoxicity of the 2,3,5,4′-tetrahydroxy-trans-stilbene-2-O-β-glucoside (trans-SG) and its cis-isomer (cis-SG). By using such an organoid platform, we successfully differentiated the two stereoisomers’ hepatotoxic potentials, which were in accordance with their differences in rodents and humans. The lesion mechanism of the toxic isomer (cis-SG) was further found as the mitochondrial injury by high-content imaging, and its hepatotoxicity could be dose-dependently inhibited by the mitochondrial protective agent. These results demonstrated the utility of the organoids-based high-content imaging approach in evaluating and predicting organ toxicity of natural products in a low-cost and high-throughput way. It also suggested the rationale to use long-term cultured organoids as an alternative toxicology platform to identify early and cautiously the hepatotoxic new drug candidates in the preclinical phase.

In the present study, we discussed the drug-induced liver injury caused by Chinese patent medicines containing Polygonum multiflorum to provide a reference for clinical rational drug use of Polygonum multiflorum and its preparations. One case of long-term taking Jingwu capsule and Huolisu oral liquid, which led to the drug-induced liver injury, was reported. The other case took Runzaozhiyang capsule for a long time, which also led to drug-induced liver injury. Jingwu capsule, Huolisu oral liquid, and Runzaozhiyang capsule all contained Polygonum multiflorum, which could result in liver injury when used long-term. Moreover, we explored the clinical features and toxicity of liver damage induced by Polygonum multiflorum. Liver damage in serum transaminase was significantly increased, and the increasing rate of ALT was more than that of AST. Jaundice appeared obviously. The liver damage mechanisms included drug metabolism, immune response, physical fitness, and many other reasons. Corresponding suggestions on rational use of Polygonum multiflorum and its preparations were presented. The dosage and period should be regulated. The index of liver function should be monitored during the medication periods. Collectively, patients with a history of liver disease or a history of allergies should pay more attention when using the above-mentioned drugs. [ABSTRACT FROM AUTHOR]

Abstract Background Diabetes is a complex metabolic disease characterized by hyperglycemia, plaguing the whole world. However, the action mode of multi-component and multi-target for traditional Chinese medicine (TCM) could be a promising treatment of diabetes mellitus. According to the previous research, the TCM of Polygonum multiflorum (PM) showed noteworthy hypoglycemic effect. Up to now, its hypoglycemic active ingredients and mechanism of action are not yet clear. In this study, network pharmacology was employed to elucidate the potential bioactive compounds and hypoglycemic mechanism of PM. Methods First, the compounds with good pharmacokinetic properties were screened from the self-established library of PM, and the targets of these compounds were predicted and collected through database. Relevant targets of diabetes were summarized by searching database. The intersection targets of compound-targets and disease-targets were obtained soon. Secondly, the interaction net between the compounds and the filtered targets was established. These key targets were enriched and analyzed by protein–protein interactions (PPI) analysis, molecular docking verification. Thirdly, the key genes were used to find the biologic pathway and explain the therapeutic mechanism by genome ontology (GO) and kyoto encyclopedia of genes and genomes (KEGG) analysis. Lastly, the part of potential bioactive compounds were under enzyme activity inhibition tests. Results In this study, 29 hypoglycemic components and 63 hypoglycemic targets of PM were filtrated based on online network database. Then the component-target interaction network was constructed and five key components resveratrol, apigenin, kaempferol, quercetin and luteolin were further obtained. Sequential studies turned out, AKT1, EGFR, ESR1, PTGS2, MMP9, MAPK14, and KDR were the common key targets. Docking studies indicated that the bioactive compounds could stably bind the pockets of target proteins. There were 38 metabolic pathways, including regulation of lipolysis in adipocytes, prolactin signaling pathway, TNF signaling pathway, VEGF signaling pathway, FoxO signaling pathway, estrogen signaling pathway, linoleic acid metabolism, Rap1 signaling pathway, arachidonic acid metabolism, and osteoclast differentiation closely connected with the hypoglycemic mechanism of PM. And the enzyme activity inhibition tests showed the bioactive ingredients have great hypoglycemic activity. Conclusion In summary, the study used systems pharmacology to elucidate the main hypoglycemic components and mechanism of PM. The work provided a scientific basis for the further hypoglycemic effect research of PM and its monomer components, but also provided a reference for the secondary development of PM.

Polygonum multiflorum Thunb. is a traditional herb which was widely distributed in China. The leaves of P. multiflorum (PML), which contains lots of antioxidant constituents, are often used for tea. In this study, three flavonoid glycosides were successfully separated and characterized from the ethyl acetate fraction of 80% ethanol extracts of PML by high-speed counter-current chromatography (HSCCC). The biphasic solvent system which can isolate purified myricitrin (44.5 mg), quercitrin (23.86 mg) and afzelin (14.39 mg) from 110 mg of the ethyl acetate fraction of PML was composed of n-hexane–ethyl acetate–methanol–water (0.5:3:1:2, v/v/v/v). Moreover, the purities of each compound were 94.9, 97.0, and 95.7% as determined by high-performance liquid chromatography (HPLC), respectively. Three flavonoid glycosides were obtained using only one-step HSCCC separation for the first time and the chemical structures of these three compounds were identified using the techniques of NMR and ESI–MS. The antioxidant activities of these three compounds, ethyl acetate fraction, and crude extract were evaluated by three antioxidant assays. The results of antioxidation assays showed that myricitrin and quercitrin have strong antioxidant activity and the two components may be the material basis of the antioxidant potential of PML. This work confirmed the feasibility of HSCCC to separate compounds from complex samples and it suggests that PML have pharmaceutical potential as natural antioxidant agents. [ABSTRACT FROM AUTHOR]

Introduction: The roots of Polygonum multiflorum (PM) serve as a classical traditional Chinese medicine (TCM), which has multiple biological activities. However, many cases of hepatotoxicity in PM have been reported in recent years. Processing PM with black beans decoction is one of the typical processing methods to reduce the hepatotoxicity of PM since ancient times. Objectives: To find potential effective constituents, as well as the optimal variety and origin of black beans for the processing of PM. Methods: Based on ultrahigh‐performance liquid chromatography Q‐Orbitrap mass spectrometry (UHPLC‐Q‐Orbitrap‐MS) analysis, we measured the contents of the two potential toxic compounds (emodin‐8‐O‐glucoside and torachrysone‐O‐hexose) in raw PM (R‐PM), PM processed with big black beans (B‐PM) and PM processed with small black beans (S‐PM). The flow cytometry method analysed the effects of different processed products of PM on apoptosis of L02 cells in different drug concentration. Proton nuclear magnetic resonance (1H‐NMR) and UHPLC‐Q‐Orbitrap‐MS together with multivariate statistical analysis were used to systematically analyse the different components between small black beans (Small‐BB) and big black beans (Big‐BB) from 30 different habitats. Results: The toxicity was ranked from small to large: S‐PM < B‐PM < R‐PM. Processing PM with black beans could significantly decrease the apoptosis rate of L02 cells, especially when the drug concentration is 80 μg/mL. Besides, we find five differential compounds (α‐arabinose, α‐galactose, proline, isomer of daidzein and isomer of genistein) may be potential active ingredients. In terms of the black beans collected from 30 producing areas, we find that Small‐BB from Weifang in Shandong province was optimum to processing PM, followed by Shangqiu in Henan province, Jilin and Liaoning province. Conclusion: The ingredients that affect the processing of PM may be attributed to α‐arabinose, α‐galactose, proline, isomer of daidzein and isomer of genistein in black beans. When the drug concentration is higher, the effect of reducing the hepatotoxicity of PM is better. Besides, Small‐BB was more effective than Big‐BB for reducing the toxicity of PM, especially Small‐BB from Weifang in Shandong, Shangqiu in Henan province and northeast China. 1H‐NMR and UHPLCQ‐Orbitrap‐MS together with multivariate statistical analysis were used to systematically analyze the different components between Small‐black beans and Big‐black beans. The ingredients that affect the processing of Polygonum multiflorum and their hepatotoxicity based on L02 cell were further investigated. Our results showed Small‐black beans was much safer and effective than Big‐black beans when was used for processing of Polygonum multiflorum. [ABSTRACT FROM AUTHOR]

Objective To determine the effect of Polygonum multiflorum extract (PME) on DNA methylation and transcriptome levels in the brain tissue of D-galactose-induced senescence C57BL/6 mice, and explore its mechanism of delaying aging. Methods Fifty C57BL/6 mice were randomly divided into 5 groups, that is, normal control group (CC), aging group (MC), high-, medium- and low-dosed PME groups (MH, MM and ML). Except the CC group, the mice of the other 4 groups were induced to establish an aging model by D-galactose. Sixty days later, the mice of the MH, MM and ML groups were given intragastrical infusion with PME at 1, 0.6 and 0.3 g·mL-1·kg-1 for 60 d, and those of CC group and the MC group received same volume of normal saline. All the mice were anesthetized, and the eyeballs were removed to collect blood samples for malondialdehyde (MDA) content, and superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activities to determine the establishment of aging model and the effect of Polygonum multiflorum in delaying aging in mice. The DNA methylation level and transcription level in brain tissue of each group of mice were detected by reduced representation bisulfite sequencing (RRBS) and RNA-seq sequencing. And bioinformatics analysis was used to analyze the differentially methylated region (DMR) and differentially expressed genes in the brain samples of each group. The related genes in the promoter region of DMR were analyzed by GO clustering and KEGG pathway analysis. Real-time qPCR was used to verify the expression of some genes. Results Compared with the CC group, the activities of SOD and GSH-Px were decreased obviously (P < 0.05), and the content of MDA was increased significantly in the MC group (P < 0.05). After PME treatment, the activities of antioxidant enzymes and MDA content in aging mice were distinctly relieved, especially in the MM group (P < 0.05). In DNA methylation sequencing, there were 52 related genes in the promoter region of DMR with methylation changes between the CC group and the MC group, and between the MC group and the MM group. Among these genes, the methylation levels of 7 genes were negatively correlated with transcriptome results, including Synpo and Mapkapk5 (associated with senescence), while the methylation levels of 13 genes were positively correlated with the transcriptome, including Tcf7 and Rab15 (associated with senescence). The mRNA levels of these 4 genes were consistent with the transcriptome sequencing results. Conclusion PME exerts a delaying senescence effect probably by regulating the methylation of Synpo, Mapkapk5, Tcf7 and Rab15 genes, and thereby regulating their transcription levels in senescent mice.

Xiao-yuan Zheng1, Shi-ming Yang2, Rong Zhang1, Su-min Wang2, Guo-bing Li1, Shi-wen Zhou1 1Department of Pharmacy, Xinqiao Hospital, Army Medical University, Chongqing, People’s Republic of China; 2Department of Gastroenterology, Xinqiao Hospital, Army Medical University, Chongqing, People’s Republic of ChinaCorrespondence: Shi-wen ZhouDepartment of Pharmacy, Xinqiao Hospital, Army Medical University, Chongqing 400037, People’s Republic of ChinaEmail zhouswxinqiao@126.comBackground: Emodin, a major component of Polygonum multiflorum (PM), has been reported to exert both protective and toxic effects in several cell types. However, the effects and underlying mechanisms of action of emodin in hepatic cells are still obscure.Methods: The present study used the normal human liver cell line L02 to investigate the effects and mechanisms of emodin in hepatic cells. After treatment with emodin, L02 cells were examined for viability, apoptosis and autophagy with the Cell Counting Kit-8 (CCK-8), annexin V/PerCP staining and GFP-LC3 plasmid transfection. The expression of proteins including cleaved caspase-3, LC3B-I/II, p-PI3K, PI3K, p-AKT, AKT, p-mTOR, mTOR and actin was examined by using Western blot.Results: Emodin significantly inhibited the viability of and induced apoptosis in L02 cells in a dose- and time-dependent manner. In addition, emodin increased the number of GFP-LC3 puncta in L02 cells and upregulated the expression of LC3B-II compared to those in control cells. Furthermore, emodin significantly decreased the expression of p-PI3K, p-AKT and p-mTOR in a dose-dependent manner compared to that in control cells without altering the expression of PI3K, AKT and mTOR. Notably, cotreatment with emodin and 3-methyladenine (3-MA) or rapamycin significantly increased and decreased the apoptosis rate of L02 cells, respectively, compared to that of cells treated with emodin alone.Conclusion: In conclusion, emodin exhibited cytotoxicity in the L02 human hepatic cell line by promoting apoptosis, and it also induced autophagy through the suppression of the PI3K/AKT/mTOR signalling pathway. The autophagy could play a protective role following emodin treatment.Keywords: Polygonum multiflorum, emodin, hepatoprotective, apoptosis, hepatotoxic, autophagy, PI3K, AKT, mTOR

Ha-Soo-Oh is a traditional Chinese medicine prepared from the roots of Polygonum multiflorum Thunb. The herb extract has been widely used in Asian countries as a tonic agent and nutritional supplement for centuries. To identify new bioactive compounds in Chinese herbs, the biotransformation-guided purification (BGP) process was applied to Ha-Soo-Oh with Bacillus megaterium tyrosinase (BmTYR) as a biocatalyst. The result showed that a major biotransformed compound could be purified using the BGP process with preparative high-performance liquid chromatography (HPLC), and it was confirmed as a new compound, 2,3,5,3′,4′-pentahydroxystilbene-2-O-β-glucoside (PSG) following mass and nucleic magnetic resonance (NMR) spectral analyses. PSG was further confirmed as a biotransformation product from 2,3,5,4′-tetrahydroxystilbene-2-O-β-glucoside (TSG) by BmTYR. The new PSG exhibited 4.7-fold higher 1,1-diphenyl-2-picrylhydrazine (DPPH) free radical scavenging activity than that of TSG. The present study highlights the potential usage of BGP in herbs to discover new bioactive compounds in the future.

The root of Reynoutria multiflora (Thunb.) Moldenke (syn.: Polygonum multiflorum Thunb., HSW) is a distinguished herb that has been popularly used in traditional Chinese medicine (TCM). Evidence of its potential side effect on liver injury has accumulated and received much attention. The objective of this study was to profile the metabolic characteristics of lipids in injured liver of rats induced by HSW and to find out potential lipid biomarkers of toxic consequence. A lipopolysaccharide (LPS)-induced rat model of idiosyncratic drug-induced liver injury (IDILI) was constructed and evident liver injury caused by HSW was confirmed based on the combination of biochemical, morphological, and functional tests. A lipidomics method was developed for the first time to investigate the alteration of lipid metabolism in HSW-induced IDILI rat liver by using ultra-high-performance liquid chromatography/Q-exactive Orbitrap mass spectrometry coupled with multivariate analysis. A total of 202 characterized lipids, including phosphatidylcholine (PC), lysophosphatidylcholine (LPC), phosphatidylethanolamine (PE), lysophosphatidylethanolamine (LPE), sphingomyelin (SM), phosphatidylinositol (PI), lysophosphatidylinositol (LPI), phosphatidylserine (PS), phosphoglycerols (PG), and ceramide (Cer), were compared among groups of LPS and LPS + HSW. A total of 14 out 26 LPC, 22 out of 47 PC, 19 out of 29 LPE, 16 out of 36 PE, and 10 out of 15 PI species were increased in HSW-treated rat liver, which indicated that HSW may cause liver damage via interfering the phospholipid metabolism. The present work may assist lipid biomarker development of HSW-induced DILI and it also provide new insights into the relationships between phospholipid perturbation and herbal-induced idiosyncratic DILI.

The goal of this research is to evaluate the influence of the polyphenols extract of Polygonum multiflorum Thunb. root on the lipid oxidation, chemical properties and sensory characteristics of ground beef during frozen storage. Beef was ground in aqueous solutions of polyphenols extract at different concentrations: 830, 415, 277, 208 and 166 mg GAE/L, polyphenols solution/sample ratio is 1/20 (v/w). Then, the ground beef was stored for up to 100 days at -20±2oC. The best oxidation inhibitor for ground beef was at the highest polyphenols concentration of 830 mg GAE/L. All quality parameters (pH, PoV, MDA, color parameter and sensory evaluation) of the treated sample and of the control sample display significant differences (p<0.05) during storage period. For this reason, it was concluded that the polyphenols extract of Polygonum multiflorum Thunb. root could be used as an alternative source of natural antioxidant in beef processing. [ABSTRACT FROM AUTHOR]

Background: Dermal 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. Methods: The 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. Results: PM 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. Conclusion: Our 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. [ABSTRACT FROM AUTHOR]

[Display omitted] • Elasticnet NIRS quantitative model of the herb Polygonum multiflorum was constructed. • The absorbance coefficient weights for each wave number were determined by the Elasticnet NIRS model. • The final coefficients may reflect the compounds of spectral feature indirectly. • The Elasticnet NIRS model showed better performance than the other four NIRS models. This study constructed a near-infrared spectroscopy (NIRS) quantitative analysis model of 2,3,5,4′-tetrahydroxy stilbene-2-O-β-D-glucoside (TSG) and Polygonum multiflorum 's water content based on Elasticnet regression (ER). NIRS data, TSG and the moisture content data sets of 117 samples of P. multiflorum were obtained by using NIR scanning and high-performance liquid chromatography analysis (HPLC) methods. MSE was used to determine the hyperparameters α and λ of Elasticnet. The optimal α values of TSG and water content were 0.83 and 0.01, and the optimal λ values were 0.03 and 1.13, respectively. By preprocessing the original spectral data, the NIRS model was established based on the source data set using ER, compared with the model established by PLS and its optimization algorithm. The results show that the ER model has good performance compared with other methods, with R2 c value was 0.9879, RMSEC value was 0.09, R2 p value was 0.9784, RMSEP value was 0.13 and RPD value was 6.8. In addition, according to the properties of Elasticnet's variable selection, we speculate that the final coefficients of spectral features may indirectly reflect the spectral features of different components. The results showed better prediction accuracy and superior performance. This is a much more reliable method used to forecast the content of effective components in plants. [ABSTRACT FROM AUTHOR]

A rapid and effective ultra-high performance liquid chromatography (UHPLC) method was developed for the determination of five anthraquinones (emodin, physcion, aloe-emodin, rhein, and chrysophanol) in Polygonum multiflorum. The target compounds were ultrasonically extracted with 70% methanol, followed by dispersive solid-phase extraction (d-SPE) with HC-C18 and desorption with acetonitrile. The five anthraquinones were separated on an ACQUITY UPLC® HSS T3 column (2.1 × 100 mm, 1.8 μm) and detected by a photodiode array detector (PDA) at 254 nm. Under the optimized conditions, linear relationships were achieved in the range of 0.3~100 mg/L for emodin, 0.3~40 mg/L for physcion, 0.1~20 mg/L for aloe-emodin, and 0.05~20 mg/L for rhein and chrysophanol. The limits of detection of the five analytes ranged from 0.01 to 0.08 mg/L, and the recoveries were within the range of 82.8~118.4% with an RSD (n = 6) of 1.0~10.3%. The intra-day and inter-day precision (n = 5) of the five targets were in the range of 1.0~1.8% and 3.0~3.1%, respectively. Furthermore, this method was applied to analyses of Polygonum multiflorum samples collected from different regions in China with satisfactory results. All the results indicated that this method is suitable for the detection of five anthraquinones in Polygonum multiflorum.

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.

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.

Polygonum multiflorum [PM, synonym Reynoutria multiflora (Thunb.) Moldenke.], a well-known and commonly used Traditional Chinese Medicine and herbal dietary supplement for nourishing the kidney and liver, etc., has aroused wide concern for its reported potential hepatotoxicity. Previous clinical cases and experimental studies have suggested that mild immune stress (MIS) may be one of the susceptibility-related factors of idiosyncratic drug-induced liver injury (IDILI) caused by PM. In this paper, we found that the same dose of PM caused abnormal liver biochemical indicators and liver tissue damage in MIS model rats, while it did not result in liver injury in normal rats, further confirming that MIS is a susceptibility factor for PM-IDILI. Plasma chemokine/cytokine profiling indicated that the MIS model group was significantly different from the other groups, showing a significant upregulation of plasma chemokines, while the MIS/PM group showed upregulated expression of chemokines or pro-inflammatory cytokines. Liver histopathological examination indicated a small amount of inflammatory cytokine infiltration in the MIS group, but no hepatocyte injury, consistent with the plasma profiles of increased chemokines and unchanged inflammatory cytokines. Notably, metabolomics characterization showed that MIS caused reprogramming of these metabolic pathways (such as phenylalanine and glutamate pathways), which was associated with acute phase reactions and inflammatory responses. These results suggested that MIS may promote an immune response to the initial cellular injury induced by PM in the liver, and MIS-induced upregulation of chemokines and metabolic reprogramming may an important mechanism that mediates the susceptibility to PM-IDILI. Furthermore, via receiver operating characteristic (ROC) curves analysis, we identified 12 plasma cytokines (e.g., IP-10, MCP-1 and MIP-1α) and nine metabolomics biomarkers (e.g., L-Phenylalanine, Creatinine, and L-glutamine) with differential capabilities (all ROC AUC > 0.9) of identifying susceptibility model animals from normal ones, which might be of referable value for the clinical recognition of PM-IDILI susceptible individuals.

The dry roots of Polygonum multiflorum (PM), involving both the raw and processed materials, are widely used as the traditional Chinese medicine for treating various diseases in China. Hepatotoxicity has been occasionally reported in patients who consume PM. Unfortunately, no definite criteria are currently available regarding the processing technology of PM for reduction the toxicity. In this work, we aimed to investigate the variations of PM metabolite profiles induced by different processing technologies by UHPLC/Q-Orbitrap-MS and multivariate statistical analysis, and to discover the potential toxic compounds by correlating the cytotoxicity of L02 cell with the contents of metabolites in raw and processed PM samples. We could identify two potential toxic compounds, emodin-8-O-glucoside and torachrysone-O-hexose, which could be selected as the toxic markers to evaluate different processing methods. The results indicated all processed PM samples could decrease the cytotoxicity on L02 cell. The best processing technology for PM process was to steam PM in black soybean decoction (BD-PM) for 24 h.

Idiosyncratic drug-induced liver injury (IDILI) is a rare but potentially severe adverse drug reaction. To date, identifying individuals at risk for IDILI remains challenging. This is a prospective study, where a nested case–control (1:5) design was adopted. For six patients who had abnormalities in liver function test after Polygonum multiflorum Thunb. (PM) ingestion (susceptible group), 30 patients with normal liver function were matched (tolerant group). Based on liquid chromatography–mass spectrometry, metabolomics analysis was done on serum samples prior to PM ingestion, to screen the differential metabolites and characterize metabolomic profiles of patient serum in the two groups. Multivariate analysis showed that there were remarkable separations between susceptible and tolerant groups. A total of 25 major differential metabolites were screened out, involving glycerophospholipid metabolism, sphingolipid metabolism, fatty acid metabolism, histidine metabolism and aromatic amino acid metabolism. Wherein, the area under the curve of the receiver operating characteristic curves of metabolites PE 22:6, crotonoyl-CoA, 2E-tetradecenoyl-CoA, phenyllactic acid, indole-5,6-quinone, phosphoribosyl-ATP were all greater than 0.9. The overall serum metabolic profile comprising of 25 metabolites could clearly distinguish susceptible and tolerant groups. This proof-of-concept study used metabolomics to characterize the metabolic profile of IDILI risk individuals before drug ingestion for the first time. The metabolome characteristics in patient serum before PM ingestion may predict the risk of liver injury after PM ingestion. [ABSTRACT FROM AUTHOR]

The roots of Polygonum multiflorum Thunb. (Vietnamese name: Ha-thu-o-do, HTOD) are used in processed form or the raw state in traditional Vietnamese medicine for many diseases and in extract form in the food industry. Some studies pointed out that HTOD extract had toxicity in humans. However, the toxicity of this herb plant currently remains unclear. In addition, this material contained a large amount of bioactive compounds, especially phenolic compounds. They have a strong antioxidant capacity and they can also interact with many different substrates such as protein, enzyme, lipid and carbohydrate. In this study, the received extracts from HTOD had the polyphenols concentrations of 415, 277, 208 and 166 (mg GAE.L-1), respectively. Besides, we only evaluated the gelatin-polyphenols interaction and the toxicity of HTOD extract in Swiss mice. The results show a strong gelatin-polyphenols interaction and no acute or subacute toxicity in mice. The polyphenols extract of HTOD at the concentration tested in this study is safe to use in food. [ABSTRACT FROM AUTHOR]

The roots of Polygonum multiflorum Thunb. (Vietnamese name: Ha-thu-o-do, HTOD) are used in processed form or the raw state in traditional Vietnamese medicine for many diseases and in extract form in the food industry. Some studies pointed out that HTOD extract had toxicity in humans. However, the toxicity of this herb plant currently remains unclear. In addition, this material contained a large amount of bioactive compounds, especially phenolic compounds. They have a strong antioxidant capacity and they can also interact with many different substrates such as protein, enzyme, lipid and carbohydrate. In this study, the received extracts from HTOD had the polyphenols concentrations of 415, 277, 208 and 166 (mg GAE.L-1), respectively. Besides, we only evaluated the gelatin-polyphenols interaction and the toxicity of HTOD extract in Swiss mice. The results show a strong gelatin-polyphenols interaction and no acute or subacute toxicity in mice. The polyphenols extract of HTOD at the concentration tested in this study is safe to use in food. [ABSTRACT FROM AUTHOR]

Objective To observe the hepatotoxicity of different extraction sites ( chloroform，ethyl acetate and residu­al aqueous phase) of polygonum multiflorum on zebrafish larvae and to explore its mechanism. Methods We weighed 1000 g polygonum multiflorum，and prepared solid extracts of chloroform，ethyl acetate and residual aqueous phase of polyg­onum multiflorum, and measured the 10% lethal dose of different extracts of polygonum multiflorum on zebrafish larvae by 24 h acute toxicity test. The dosages of different extraction sites of polygonum multiflorum in the subsequent experiments were determined. The low dose, medium-dose and high-dose chloroform were 3,6, and 9 |jig/mL, respectively. The low- dose，medium-dose and high-dose ethyl acetate were 10，20，and 30 jjig/mL，respectively. The low, medium and high do- ses of the remaining aqueous phase were 20，40，and 60 jjig/mL, respectively. The zebrafish larvae were divided into groups a，b，c and d. The high-dose chloroform extract was added to the group a，ethyl acetate extract was added to the group b，residual water extract was added to the group c，and the same amount of culture water was added to the group d. After 24-hour incubation, the pathological changes of liver of each group were observed under the microscope, and the liver-body ratio and yolk sac ratio were calculated. The zebrafish larvae were divided into the groups Al，A2，A3，A4， B1，B2，B3, B4，Cl，C2, C3, and C4. The low dose，medium-dose and high-dose chloroform extracts were added to the groups Al，A2 and A3; the low-dose，medium-dose and high-dose ethyl acetate extracts were added to the groups B1，B2 and B3 ï¼› and the low-dose，medium-dose and high-dose residual water extracts were added to the groups Cl，C2 and C3, respectively, and the same amount of culture water were added to the groups A4，B4，and C4，respectively. The ALT and AST of zebrafish larvae in each group were detected by Reiss method, and T-SOD of zebrafish larvae in each group was detected by hydroxylamine method. Results The liver of zebrafish larvae in the groups a，b，c and d was transparent, clear and intact. The liver of zebrafish larvae in the groups a，b，c and d became dark grey, with de­creased transparency and enlarged liver. The liver-body ratios of zebrafish larvae in the groups a，b，c and d were 9.631% ±0.011% , 10.061% ±0.014% , 8.235% ±0.010% , and 7.785% ±0.013% , respectively. The yolk sac percentage of zebrafish larvae in the groups a, b, c and d were 3. 679% ±1.131%，3. 648% ± 1.388%，3. 862% 土1. 345% and 3. 893% ±0.601%，respectively, with no statistically significant difference ( all P > 0. 05 ) . Compared with the group d，the number of hepatocytes in the groups a and b decreased, and there were vesicular degeneration and irregular arrangement, indicating the liver injury. The ALT levels of the groups Al，A2，and A3 were significantly higher than those of the group A4 ( P < 0. 05 ) , the groups B1，B2，and B3 were significantly higher than group B4 ( P <0. 05 )，and the groups C2 and C3 were significantly higher than the group C4 group ( P < 0. 05 ) . The AST levels of the group A3 was significantly higher than that of the group A4 ( P < 0. 05 ) , and the group B3 was significantly higher than the group B4 ( P < 0. 05 ) . T-SOD level in the group A3 was significantly higher than that in the group A4 ( P < 0. 05 ), and those in the groups B1，B2 and B3 were significantly higher than that in the group B4 ( all P < 0. 05 ) . Conclusions The chloroform and ethyl acetate fractions of polygonum multiflorum have hepatotoxicity to zebrafish larvae. The mech­anism may be that the toxic components of polygonum multiflorum destroy the balance of oxidative stress in zebrafish, and then lead to hepatocyte damage. [ABSTRACT FROM AUTHOR]

A vast array of plant-based compounds has enriched red biotechnology to serve the human health and food. A peculiar medicinal plant which was an element of traditional Chinese medicine for centuries as a liver and kidney tonic, for life longevity and hair blackening, is Polygonum multiflorum Thunb. (PM) which is popularly known as "He shou wu" or "Fo-ti" and is rich in chemical components like stilbenes, quinones, and flavonoids which have been used as anti-aging, anti-alopecia, anti-cancer, anti-oxidative, anti-bacterial, anti-hyperlipidemia, anti-atherosclerosis, and immunomodulating and hepatoprotective agents in the modern medicine. The health benefits from PM are attained since long through commercial products such as PM root powder, extract, capsules, tincture, shampoo, and body sprays in the market. Currently, the production of these pharmaceuticals and functional foods possessing stilbenes, quinones, and flavonoids is through cell and organ cultures to meet the commercial demand. However, hepatotoxic effects of PM-based products are the stumbling blocks for its long-term usage. The current review encompasses a comprehensive account of bioactive compounds of PM roots, their biological activities as well as efficacy and toxicity issues of PM ingredients and future perspectives. [ABSTRACT FROM AUTHOR]

The dry roots of Polygonum multiflorum (PM), involving both the raw and processed materials, are widely used as the traditional Chinese medicine for treating various diseases in China. Hepatotoxicity has been occasionally reported in patients who consume PM. Unfortunately, no definite criteria are currently available regarding the processing technology of PM for reduction the toxicity. In this work, we aimed to investigate the variations of PM metabolite profiles induced by different processing technologies by UHPLC/Q-Orbitrap-MS and multivariate statistical analysis, and to discover the potential toxic compounds by correlating the cytotoxicity of L02 cell with the contents of metabolites in raw and processed PM samples. We could identify two potential toxic compounds, emodin-8- O -glucoside and torachrysone- O -hexose, which could be selected as the toxic markers to evaluate different processing methods. The results indicated all processed PM samples could decrease the cytotoxicity on L02 cell. The best processing technology for PM process was to steam PM in black soybean decoction (BD-PM) for 24 h. [ABSTRACT FROM AUTHOR]

Polygonum multiflorum [PM, synonym Reynoutria multiflora (Thunb.) Moldenke.], a well-known and commonly used Traditional Chinese Medicine and herbal dietary supplement for nourishing the kidney and liver, etc., has aroused wide concern for its reported potential hepatotoxicity. Previous clinical cases and experimental studies have suggested that mild immune stress (MIS) may be one of the susceptibility-related factors of idiosyncratic drug-induced liver injury (IDILI) caused by PM. In this paper, we found that the same dose of PM caused abnormal liver biochemical indicators and liver tissue damage in MIS model rats, while it did not result in liver injury in normal rats, further confirming that MIS is a susceptibility factor for PM-IDILI. Plasma chemokine/cytokine profiling indicated that the MIS model group was significantly different from the other groups, showing a significant upregulation of plasma chemokines, while the MIS/PM group showed upregulated expression of chemokines or pro-inflammatory cytokines. Liver histopathological examination indicated a small amount of inflammatory cytokine infiltration in the MIS group, but no hepatocyte injury, consistent with the plasma profiles of increased chemokines and unchanged inflammatory cytokines. Notably, metabolomics characterization showed that MIS caused reprogramming of these metabolic pathways (such as phenylalanine and glutamate pathways), which was associated with acute phase reactions and inflammatory responses. These results suggested that MIS may promote an immune response to the initial cellular injury induced by PM in the liver, and MIS-induced upregulation of chemokines and metabolic reprogramming may an important mechanism that mediates the susceptibility to PM-IDILI. Furthermore, via receiver operating characteristic (ROC) curves analysis, we identified 12 plasma cytokines (e.g., IP-10, MCP-1 and MIP-1α) and nine metabolomics biomarkers (e.g., L-Phenylalanine, Creatinine, and L-glutamine) with differential capabilities (all ROC AUC > 0.9) of identifying susceptibility model animals from normal ones, which might be of referable value for the clinical recognition of PM-IDILI susceptible individuals. [ABSTRACT FROM AUTHOR]

The correlation between growth and accumulation of bioactive compounds was investigated in six genotypes of Polygonum multiflorum, grown in vitro, soil-grown (ex vitro), and as adventitious root cultures. Adventitious roots were induced on leaf and root explants from plantlets of the six genotypes grown in vitro. Line PM-06 achieved the highest biomass in vitro (0.91 g plant−1 FW; 0.1 g plant−1 (DW)), whereas line PM-05 produced the highest biomass ex vitro (54.67 g plant−1 FW; 12.93 g plant−1 DW). A comparison of the six lines found that total phenolic content (TPC) and total flavonoid content (TFC) were highest in roots from line PM-01 grown in vitro, but line PM-06 produced the highest levels of bioactive compounds in tubers (99.06 mg g−1 DW TPC; 42.31 mg g−1 DW TFC), which were 2.5-fold, 5.0-fold, and 4.8-fold higher than the highest levels produced by in vitro roots, in vitro shoots, and ex vitro shoots, respectively. Although adventitious root line AR-06 produced the greatest overall biomass (60.12 g L−1 FW; 6.36 g L−1 DW), bioactive content was highest in line AR-01 (50.35 mg g−1 DW TPC; 22.51 mg g−1 TFC). There was a strong correlation between phenolic production in plant roots grown in vitro and adventitious root lines, and plant roots grown ex vitro and adventitious root lines. Adventitious root cultures of P. multiflorum line AR-06 showed great potential for producing phenolic compounds. Such cultures may therefore provide an alternative to naturally grown plants as a potential biomass source for the production of bioactive compounds. [ABSTRACT FROM AUTHOR]

Graphical abstract Highlights • The bioassay method could be used to evaluate the quality of Polygonum multiflorum associated with antiplatelet aggregation effects. • Chemical fingerprints revealed variations in the contents of seven main peaks contained in 10 batches of Polygonum multiflorum. • Trans -SG and catechin might be the active constituents of antiplatelet aggregation of Polygonum multiflorum. Abstract Currently, an increasing number of patients are seriously affected by acute thrombotic events. In China, Polygonum multiflorum (PM) is commonly used to treat diseases associated with thrombosis. Our previous work showed that PM could inhibit the platelet aggregation that plays a key role in the pathogenesis of thrombosis. However, the constituents of PM are complicated, and quality control methods cannot completely ensure the quality and clinical efficacy. In an attempts to explore this problem, we constructed a direct bioassay method to evaluate the antiplatelet aggregation effects of PM. To ensure the precision and reliability of this bioassay, we optimized and standardized the experimental conditions and then tested the standardized bioassay by analyzing 10 PM samples. Additionally, we combined chemical and biological evaluation methods to identify antiplatelet aggregation markers. The evaluation indicated that 10 samples of PM could inhibit platelet aggregation and there was a notable difference in biopotency between the different PM groups. Chemical fingerprints revealed variations in the contents of the 7 main peaks. Trans -2,3,5,4′-tetrahydroxy-stilbene-2-O-β- d -glucoside and catechin might be active constituents of antiplatelet aggregation as determined by spectrum-effect relationships. This work indicates that bioassay and spectrum-effect relationships are useful tools to associate sample quality with the potential chemical markers linked to the clinical effects of Traditional Chinese Medicines. [ABSTRACT FROM AUTHOR]

Graphical abstract Highlights • Characteristics to distinguish F. multiflora , F. multiflora var. angulata , and F. multiflora var. ciliinervis were shown. • There were significant differences in chemical compositions among the three taxa. • F. multiflora var. ciliinervis is an independent species. • F. multiflora var. angulata should be classified as a variety of F. multiflora. Abstract Relationships among Fallopia multiflora (Thunb.) Haraldson., F. multiflora var. angulata (S. Y. Liu) H. J. Yan, Z. J. Fang & Shi Xiao Yu., and F. multiflora var. ciliinervis (Nakai) Yonekura & H. Ohashi. were determined based on macroscopic and microscopic morphology, molecular phylogeny, and chemical analysis. The macroscopic and microscopic morphologies of root tubers or rhizomes, stems, and leaves were compared among the three taxa. The content of 11 chemical components (catechin, polydatin, stilbene glucoside, emodin, emodin-8- O -β-D-glucopyranoside, rhein, chrysophanol, aloe-emodin, quercetin, physcion, and resveratrol) in the three taxa was determined by HPLC, and the chemical diversity was further evaluated by principal component and hierarchical cluster analyses. Molecular phylogenies were mapped using two chloroplast markers (matK and the psbA-trnH intergenic region) and a nuclear ribosomal marker [internal transcribed spacer 2 (ITS2) region]. Analyses of macroscopic and microscopic morphological characteristics revealed that the subterranean organs of F. multiflora and F. multiflora var. angulata are root tubers, whereas those of F. multiflora var. ciliinervis are rhizomes. In the phylogenetic trees, F. multiflora and F. multiflora var. angulata were clustered into a clade based on the combine matK + psbA-trnH sequence, with neighbour-joining, maximum likelihood, and Bayesian inference bootstrap support values of 99, 85, and 0.99, respectively. In addition, there were obvious differences in the chemical compositions of F. multiflora , F. multiflora var. angulata and F. multiflora var. ciliinervis. The root tubers of F. multiflora contain higher levels of stilbene glucoside and catechin, but lower levels of polydatin and anthraquinone compounds. In contrast to F. multiflora , the rhizomes of F. multiflora var. ciliinervis contain higher levels of polydatin and anthraquinone compounds, but lack stilbene glucoside. The content of all 11 assessed components was lower in F. multiflora var. angulata than in F. multiflora and F. multiflora var. cillinervis. Principal component and hierarchical cluster analyses revealed that F. multiflora and F. multiflora var. angulata individuals were clustered into a single clade, whereas F. multiflora var. ciliinervis individuals were clustered into a single clade separate from that containing F. multiflora and F. multiflora var. angulata individuals. On the basis of the results of our morphological, molecular phylogeny, and chemical analyses, we tentatively conclude that F. multiflora var. ciliinervis is an independent species, whereas F. multiflora var. angulata should be considered as a variety of F. multiflora. [ABSTRACT FROM AUTHOR]

Emodin is an active ingredient in many herbal medicines and has a broad spectrum of pharmacological activities. The current data indicate that emodin exerts its beneficial effect on alpha-naphthylisothiocyanate (ANIT)-induced intrahepatic cholestasis through its anti-oxidant and anti-inflammatory activities. Emodin has little effect on the concentrations of bile acids (BAs) in livers of ANIT-treated mice. Instead, emodin shows a potential pro-cholestatic effect by interfering with the crosstalk between AMP-activated protein kinase (AMPK) and farnesoid X receptor (Fxr) in the liver, which leads to a suppression of bile salt export pump (Bsep). Two emodin-containing herbs, namely Polygonum multiflorum (PM) and Semen cassiae (SC), markedly aggravate the intrahepatic cholestasis in ANIT-treated mice. SC interferes with the AMPK-Fxr crosstalk and suppresses Bsep in livers of mice. ANIT markedly increases the hepatic retention of emodin in SC-treated mice. The major SC constituents, in particular three anthraquinones, are able to activate AMPK in HepG2 cells and inhibit Bsep in primary mouse hepatocytes, with emodin showing the strongest activities. Together, the present study identifies a potential pro-cholestatic role of emodin in the hepatotoxicity of herbs. [ABSTRACT FROM AUTHOR]