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

To standardize the processing technology of Polygoni Multiflori Radix Praeparata and stabilize its quality, the similar change laws of Polygoni Multiflori Radix Praeparata with different processing methods and time were analyzed. The processing time of Polygoni Multiflori Radix Praeparata was studied at 24, 32, 40, 48 h, and 4 different processing methods were studied, namely stewed with black bean sauce, steamed, steamed with black bean sauce, and steamed with black bean sauce and rice wine. The content of stilbene glycosides and anthraquinones were determined by HPLC-DAD method. UV method was used to determine the content of polysaccharides, and HPLC-ELSD method was used to determine the monosaccharides and oligosaccharides. The comparative chart of content determination, cluster analysis and entropy weight TOPSIS model was used to find the similar change laws and time interval of different processing methods of Polygoni Multiflori Radix Praeparata. The results demonstrated that around 32 h, the content of nine components in Polygoni Multiflori Radix Praeparata with different processing methods had similar change laws, and the decoction pieces had a high quality, indicating that the four processing methods of Polygoni Multiflori Radix Praeparata are likely to be used as one type of decoction piece with the same name.
Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; Glycosides ; Plant Roots ; Polygonum

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

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

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

In this study, we aimed to establish a rat liver micro-tissue evaluation system to evaluate the hepatotoxicity of the main monomers in Polygonum multiflorum. Rat primary hepatocytes were isolated and purified by two-step in situ perfusion method to prepare hepatic parenchymal cells. The ultra-low adsorption plate and the inverted model were used to establish an in vitro hepatotoxicity evaluation system. After the system was established, the main monomer components(monanthone with emodin type, rhein, emodin, emodin-8-O-β-D-glucopyranoside, physcion) of P. multiflorum were selected for in vitro hepatotoxicity evaluation. This study showed that the primary cells of the liver can form liver micro-tissues in the low adsorption plate method and the mold perfusion method, with good liver structure and function, which can be used to evaluate the hepatotoxicity of the drug to be tested after long-term administration. The five monomers to be tested in P. multiflorum can significantly affect the proliferation of primary liver micro-tissues in rats in a dose-and time-dependent manner. The hepatotoxic effects were as follows: monanthone with emodin type > rhein > emodin > emodin-8-O-β-D-glucopyranoside > physcion. The results suggested that the emodin-type monoterpene and rhein might be the potential hepatotoxic components, while the metabolites of emodin-8-O-β-D-glucoside and emodin methyl ether showed more toxic risks. The rat primary hepatocyte micro-tissue model system established in this experiment could be used to achieve long-term drug administration in vitro, which was consistent with the clinical features of liver injury caused by long-term use of P. multiflorum. The experimental results provided important information and reference on the clinical application and toxic component of P. multiflorum.
Animals ; Chemical and Drug Induced Liver Injury ; Emodin ; Fallopia multiflora ; Glucosides ; Plant Extracts ; Polygonum ; Rats

Polygoni Mulitiflori Radix, or dried root tuber of Polygonum multiflorum(PM), is a traditional Chinese tonic medicine, with the effect in nourishing liver and kidney, and benefiting blood essence and hair. It is widely used in clinical and healthcare products. In recent years, more and more reports about adverse reactions of root tuber of P.multiflorum and its preparations have been reported. Fortunately, there is also substantial progress in the experimental study on liver injury induced by PM. According to the literature review, the possible causes of liver injury were found to be the mixture of raw and processed PM and long-term high-dose administration. In addition, the liver injury induced by PM is idiosyncratic liver injury, and individual factors are also the important cause. At the same time, according to the literature reports, the effects of chemical components in different pathological animal models were summarized, finding that 2,3,5,4'-tetrahydroxystilbene-2-O-β-D-glucoside was the main component for liver injury; based on the clinical manifestations of liver injury induced by PM, the effects of some chemical components on bilirubin and bile acid metabolism were analyzed. This paper reviews the study progress of liver injury induced by PM.
Animals ; Chemical and Drug Induced Liver Injury ; Drugs, Chinese Herbal ; Plant Roots ; Polygonum

Ultra performance liquid chromatography coupled with time-of-flight mass spectrometry( UPLC-Q-TOF-MS/MS) method was applied to analyze the prototypes and metabolites of the effective components of Polygonum orientale in SD rat serum and urine. The separation was performed on Agilent Eclipse Plus C_(18) column( 2. 1 mm×100 mm,1. 8 μm),with 0. 1% formic acid solution( A)-acetonitrile( B) as the mobile phase for gradient elution. Mass spectrometry data of biological samples were obtained under positive and negative electrospray ion mode. By comparing chromatogram differences between blank samples and drug treatment samples,prototype components and metabolites of the effective components of P. orientale extract were identified. The results showed that 12 metabolites were detected in serum and 26 metabolites in urine( including cross-components) of rats. The main metabolic pathways included hydrogenation,hydroxylation,glucuronidation,sulfation reaction,and methylation-glucuronidation,etc. The method established in this study was reliable and effective for studying the metabolic characteristics of the effective components of P. orientale in rats,and it can provide a reference for further studies on therapeutic material basis of this herb.
Animals ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; pharmacokinetics ; Flowers ; chemistry ; Phytochemicals ; blood ; urine ; Polygonum ; chemistry ; Rats ; Rats, Sprague-Dawley ; Tandem Mass Spectrometry

To explore the immune status of patients with drug-induced liver injury caused by Polygonum multiflorum preparations,and analyze their immune characteristics. Case-control design was used to collect the cases of drug-induced liver injury caused by P. multiflorum preparations through key specialized surveillance. Five matching factors,namely type of P. multiflorum preparations,gender,age,basic diseases and concomitant medication were controlled. According to the ratio of 1 ∶ 1,cases of patients who took P. multiflorum preparations but with no liver injury were monitored at prospective hospitals. The demographic information,disease information,medication information and laboratory examination information of the two groups were recorded,and venous blood was collected. The gene sequence was detected by high-throughput sequencing technology,and the characteristics of TCR immune repertoire of the two groups were analyzed. A total of 46 pairs of patients were enrolled in the study. The results showed significant differences in the number of CDR3 and clone species,the length of amino acid sequence in CDR3 region,the abundance of V gene and J gene,the cross-linking of V-J gene and the diversity of immune repertoire between patients with drug-induced liver injury and patients without liver injury. The immunohistochemical diversity and high-frequency V-J cross-linking characteristics of patients with liver injury caused by P. multiflorum preparations were found,which provided a reference for screening out drug users to reduce the occurrence of liver injury caused by P. multiflorum preparations.
Chemical and Drug Induced Liver Injury ; Drugs, Chinese Herbal/adverse effects* ; Fallopia multiflora ; Humans ; Polygonum ; Prospective Studies ; Receptors, Antigen, T-Cell

Based on the ancient method of nine-steaming and nine-sun-curing,the chemical composition changes and quality profiles in different processes of Polygoni Multiflori Radix were studied. Their contents of stilbene glycoside,anthraquinones and polysaccharides were determined by nine-steaming and nine-sun-curing with black bean juice and pharmacopoeia method. HPLC chemical fingerprints were established,and orthogonal partial least squares-discriminant analysis( OPLS-DA) was performed on different processed products using SIMCA 14. 1 software to evaluate the quality difference between samples. The results of content determination show that,with the increase of the number of processing and steaming times,the stilbene glycoside and the combined anthraquinone showed a decreasing trend,and the free anthraquinone,total anthraquinone and polysaccharide showed an upward trend in the different preparations of Polygoni Multiflori Radix and Pharmacopoeia. Six-steamed and six-sun-cured products can be used as the finishing point for the classic steaming. Fingerprint results showed that there were significant differences in chemical composition in Polygoni Multiflori Radix at different processing processes. It can be identified stilbene glycoside( peak 13),emodin( peak 21),and physcion( peak 24). By comparing the relative peak areas of the 26 chromatographic peaks in the sample after normalization( the reference is peak 7),it was found that the relative peak areas of 12 peaks in the processed products were higher than the raw products,13 peaks were reduced; according to statistical analysis of OPLS-DA,Polygoni Multiflori Radix at different processing degrees was further divided into three categories,sample S1 was class I,S2-S5 were class Ⅱ,and S6-S11 were class Ⅲ． And 8 peaks with the VIP value higher than 1. 0 were peak 13,21,4,3,11,14,5,and 24 in order. The eight chemical components were the main components to distinguish the difference between Polygoni Multiflori Radix in the process of nine-steaming and nine-sun-curing,suggesting that it was rational to use stilbene glycoside,emodin and emodin methyl ether as quality control indicators of Polygoni Multiflori Radix. The method established in this experiment conformed to the methodological verification requirements,established a method of multi-component content determination combined with fingerprint,and clarified that six-steaming and six-sun-curing was used as an improved classical processing technology,and more clearly defined the whole dynamic change of chemical composition in Polygoni Multiflori Radix by nine-steaming and ninesun-curing process. It provides a basis for the chemical quality evaluation model about different processed products of Polygoni Multiflori Radix.
Anthraquinones/analysis* ; Chromatography, High Pressure Liquid ; Discriminant Analysis ; Drugs, Chinese Herbal/chemistry* ; Glycosides/analysis* ; Least-Squares Analysis ; Phytochemicals/analysis* ; Plant Roots/chemistry* ; Polygonum/chemistry* ; Polysaccharides/analysis* ; Steam ; Stilbenes/analysis* ; Technology, Pharmaceutical/methods*