Study On Hepatotoxicity Mechanism Of Chinese Herbal Medicine Gynura Segetum Based On Metabolomics

The traditional Chinese medicine Gynura segetum(Gynura segetum(Lour.)Merr.)is an herb that has the functions of promoting blood circulation,removing blood stasis,and stopping bleeding.It contains pyrrolidine alkaloids(PAs).High-dose and long-term use of Gynura segetum may cause severe liver toxicity such as ascites,hepatomegaly,and jaundice.Since the hepatotoxicity mechanism of Gynura segetum is not yet fully understood,it is difficult to diagnose and identify clinically,especially the early diagnosis biomarkers of hepatotoxicity.Therefore,this study analyzed the changes of the endogenous metabolites in the body caused by Gynura segetum induced liver poisoning,and clarified the overall effect of Gynura segetum decoction on the body and revealed its liver from the perspective of metabonomics.The potential markers of hepatotoxicity were excavated in the course of mechanism study to provide basis for rapid diagnosis,prognosis assessment and safe drug use for clinical Gynura segetum liver poisoning.It also provides ideas and evidence for the development of new drugs that cause liver damage after Gynura segetum causes liver poisoning.1.In this study,we observed the hepatic pathological changes of rats fed with Gynura segetum decoction at low-,middle-,and high-concentration groups,and established an animal model for simulating the hepatotoxicity-induced HSOS induced by Gynura segetum in clinical patients.Observe and record changes in the signs of rats during administration.After the end of administration,perform blood biochemical tests and pathological examinations of liver specimens.Determine the lesions and severity according to the Deleve score criteria under light microscope.The results showed that the pathological features of HSOS were observed after continuous administration of Gynura segetum decoction at low,middle and high concentration groups,and the model was successful.The hepatotoxicity symptoms were more obvious with the increase of rats' dose.2.The UPLC/MS technique was used to obtain the profile of metabolites in plasma,urine,and liver tissues of rat liver poisoning induced by Gynura segetum at different time points and different doses.Multivariate statistical analysis methods such as principal component analysis and partial least squares discrimination were used.Analytical methods and orthogonal partial least squares methods were used to establish a metabolomics hepatic toxicity model of plasma,urine,and liver tissue samples from normal group and different time points and different doses of hepatotoxicity group.The results showed that plasma and urine of rats in the administration group The cluster distribution of the phenotypes of fluid and liver metabolites deviated from the control group,whereas the phenomenon that the metabolites in the administration group"deviated" from the normal group was exactly the expression of liver toxicity.With the increase of the administration time,the administration was performed.The "degree" of the group"deviating" from the normal group was even more pronounced,indicating that long-term administration of Gynura segetum decoction can cause significant hepatotoxicity damage in rats,and the degree of damage is positively correlated with the content of Gynura segetum decoction.The hepatotoxicity caused by decoction of Gynura segetum in different concentration groups was evaluated from the overall level.3.Multivariate statistical analysis of endogenous metabolites in the blank group and the model group yielded differential metabolites between the normal group and the model group,and further identified and screened the liver toxicity caused by Gynura segetum decoction.The 16 potential biomarkers were L-arginine,Valine,Glutamic acid,L-proline,Linoleic acid,arachidonic acid,Pyrroline hydroxycarboxylic acid,Phytosphingosine,L-tryptophan,Sphingosine,Sphingosine 1-phosphate,L-Phenylalanine,Creatine,Sphingosine,Citric acid and LPC(4 species),as well as the associated 12 metabolic pathways:D-glutamine and D-glutamate metabolism,Alanine,aspartate and glutamate metabolism,Arginine and proline metabolism,Glyoxylate and dicarboxylate metabolism,Phenylalanine,tyrosine and tryptophan biosynthesis,Phenylalanine metabolism,Valine,leucine and isoleucine biosynthesis,Arachidonic acid metabolism,Tryptophan metabolism,Linoleic acid metabolism,Glycerophospholipid metabolism and sphingolipid metabolism.The analysis of metabolic pathways and metabolic network attribution of Gynura segetum biomarkers explored the toxicological targets and toxic mechanisms of Gynura segetum and revealed that Gynura segetum can cause disorders of metabolic pathways such as amino acid metabolism,fat metabolism,and energy metabolism,from the metabolome.From an academic point of view to further elucidate the hepatotoxicity mechanism of Gynura segetum.