Discussion On Therapeutic Mechanism Of Geniposide For Cholestasis

BackgroundGardenia jasminoides Eills is the dried and mature fruit of Rubiaceae gardenia,which has the functions of purging fire and eliminating troubles,clearing heat and diuresis,cooling blood and detoxification.Geniposide(GE)is the main active ingredient isolated from the fruit of gardenia and plays an important role in the prevention and treatment of liver diseases.Intrahepatic cholestasis(IC)is one of the liver’s common diseases,and more and more attention has been paid to the research and development of drugs for this disease.Cholestatic liver disease is caused by a variety of causes of bile production,secretion,excretion of obstacles,because the initiative can not be discharged through the bile duct to the intestinal cavity,in the liver accumulation,back into the blood,thus caused a series of organic damage,functional disorders,metabolic disorders of the liver system.Traditional Chinese medicine is becoming more and more critical in treating liver injury,but it is challenging to elucidate its mechanism and target by traditional methods because of its complex components.ObjectiveTo evaluate the efficacy of Geniposide in anti-cholestasis by establishing a rat cholestatic liver injury model model,using metabonomics and transcriptomics technology(RNA-seq)for analysis experiments,from the perspective of metabolomics and transcriptomics Explain the mechanism of Geniposide in the treatment of cholestatic liver injury.Methods(1)Pharmacodynamic analysis methodThe protective effects of Geniposide on rats with intrahepatic cholestasis induced by ANIT were investigated by investigating the changes of multiple biochemical indexes in serum and liver tissue of cholestasis model rats and liver tissue pathological slices with different doses of Geniposide.Transcriptomics analysis method(2)Metabolomics analysis methodThe rat cholestasis liver injury model was established with ANIT(α-naphthalene isothiocyanate)olive oil solution,and the level of inflammatory factors in the biochemical indicators after administration was measured to illustrate the effect of Geniposide on cholestasis liver injury.Protective effects.Results(1)Geniposide obtained by pharmacodynamic methods can reduce the serum aspartate aminotransferase(AST),alanine aminotransferase(ALT),alkaline phosphatase(AKP),and direct bilirubin(DBIL),total bile acid(TBA)and total bilirubin(TBIL)activity,increase superoxide dismutase(SOD),glutathione activity(GSH),reduce malondialdehyde(MDA)concentration,interleukin 6(IL-6),interleukin 1β(IL-1β),tumor necrosis factor α(TNF-α)levels.In addition,hematoxylin-eosin(HE)staining showed that low,medium and high doses of Geniposide can improve inflammatory cell infiltration and liver cell damage to varying degrees,and there is a dose-effect relationship.This study proved that Geniposide has a certain protective effect in IC-type injury,and its mechanism of action may be related to improving the antioxidant capacity of liver tissue and inhibiting the excessive secretion of cytoinflammatory factors.(2)Through the study of transcriptomics method,compared with the blank group,the gene comparison analysis after the ANIT modeling has screened a total of 2253 differential genes,including 1283 up-regulated genes and 970 down-regulated genes.Compared with the model group,739 differential genes were screened after Geniposide treatment,including 338 up-regulated genes and 401 down-regulated genes.GO function enrichment analysis showed that the gene function after ANIT modeling is mainly related to cholesterol-like process,lipid oxidation and neutrophil chemotaxis,and Geniposide is mainly related to organic anion transport,steroid metabolism process and cells after administration of Geniposide.Lipid catabolism is related to the process.KEGG is mainly concentrated in retinol metabolism,steroid hormone synthesis,linoleic acid metabolism,PPAR signaling pathway and bile secretion pathway.(3)A total of 98 differential metabolites were screened through the study of metabolomics methods,through the use of T-test combined with multivariate analysis of OPLS-DA.After the intervention of Geniposide,57 metabolites were up-regulated and 41 metabolites levels were up-regulated.Down.The cluster analysis of differential metabolites showed that the model group and the drug group were divided into two categories without supervision,which was consistent with the results of OPLS-DA analysis.Through enrichment analysis of the KEGG pathway,9 important metabolic pathways were screened,showing that the intervention of Geniposide in the rat intrahepatic cholestasis model may be related to fatty acid degradation,bile secretion,glutathione metabolism,neomycin,kanamycin The biosynthesis of gentamicin and gentamicin,histidine metabolism,ABC transporter,galactose metabolism,primary bile acid biosynthesis and pentose phosphate pathway are closely related.After combined analysis of metabolomics and transcriptomics,the common pathways obtained are the bile secretion pathway and the glutathione metabolism pathway.After the GE group was administered,the bile secretion pathway was enriched with 10 differential genes and 4 differences.Metabolites;9differential genes and 2 differential metabolites are enriched in the glutathione pathway.The Western blot method was used to detect and analyze the protein expression levels of the top ten targets of molecular docking.The results showed that:in this study,compared with the blank solvent group,CEH1,OAT3,ABCG5,GST,KCNN2 after modeling Protein expression levels were down-regulated,and NROB2 abnormally increased,and Geniposide was significantly adjusted after the intervention of Geniposide,suggesting that the anti-cholestosis effect of Geniposide is inseparable from its regulation of CEH1,OAT3,ABCG5,KCNN2 and NROB2 proteins in the bile secretion pathway and GST protein in the glutathione pathway.Conclusion:Geniposide has a certain protective effect on IC liver injury caused by ANIT.Its protective mechanism may be related to its ability to regulate the protein expression levels of OAT3,ABCG5,GST,CEH1,KCNN2 and NROB2,thereby regulating the bile secretion pathway and Glutathione pathway,strengthening the body’s ability to resist oxidative stress is related to inhibiting the secretion of inflammatory factors.