| Diabetes mellitus type 2 is a common endocrine metabolic disorder disease,often accompanied by hyperlipidemia,and its incidence is increasing year by year and showing a trend of younger age.The long-term use of western medicine to treat diabetes mellitus type 2 combined with hyperlipidemia in clinic can cause adverse reactions such as hypoglycemia,liver function injury and skin rash.Traditional Chinese medicine(TCM)has become a potential drug to treat type 2 diabetes because of its multiple targets and low side effects.Compatibility of Panax notoginseng,Salvia miltiorrhiza and Crataegus pinnatifida Bunge has been reported to have hypolipidemic and hypoglycemic effects.However,its mechanism of action is not yet clear.In this paper,HepG2 cell model was used to screen the optimal compatibility ratio of Sanqidansenshanzha complex prescription for reducing lipid.The lipid accumulation model and insulin resistance model of HepG2 cells were used to evaluate the lipidreducing and hypoglycemic activities of Sanqidansenshanzha complex prescription in vitro.Meanwhile,the reducing blood lipid and hypoglycemic activities in vivo of the Sanqidansenshanzha complex prescription were investigated in a mouse hyperlipidemia model induced by high-fat diet and a mouse model of diabetes mellitus type 2 induced by high-glucose and high-fat diet combined with streptozotocin.Through network pharmacology analysis,the potential targets and related pathways of Sanqidansenshanzha complex prescription for the treatment of diabetes mellitus type 2combined with hyperlipidemia were screened out.Based on metabonomics technology,the effect of Sanqidansenshanzha complex prescription on the changes of fecal metabolites in hyperlipidemic mice was investigated.Based on metabonomics technology,the effects of Sanqidansenshanzha complex prescription on the changes of fecal metabolites in hyperlipidemic mice were investigated,and the mechanism of Sanqidansenshanzha complex prescription on reducing blood lipid and hypoglycemic activities was further elucidated from the perspective of bile acid metabolism,providing theoretical basis for the clinical application of Sanqidansenshanzha complex prescription.The main research content and results are as follows:1.Sanqidansenshanzha complex prescription has in vitro reducing lipid and hypoglycemic effects.HepG2 cells were used to establish a lipid accumulation model to screen the optimal ratio of Sanqidansenshanzha complex prescription.When the compatibility ratio of Sanqidansenshanzha complex prescription was 3:2:1,the complex prescription had the best effect on reducing blood lipid.The optimal compatibility of Sanqidansenshanzha complex prescription reduced lipid accumulation and decreased intracellular triglyceride(TG)content in HepG2 cells.To establish the model of insulin resistance in HepG2 cells,the optimal compatibility of Sanqidansenshanzha complex prescription reduced the content of glucose in the cell supernatant and increased the utilization and uptake of glucose by cells.2.Sanqidansenshanzha complex prescription has reducing blood lipid and hypolipidemic effects in vivo.The high-fat diet induced hyperlipidemia model in mice was established to evaluate the in vivo hypolipidemic activity of Sanqidansenshanzha complex prescription.The results showed that Sanqidansenshanzha complex prescription reduced the liver index in mice,reduced the contents of total cholesterol(TC),triglyceride(TG)and low-density lipoprotein(LDL-C),increased the high-density lipoprotein(HDL-C)levels in mouse plasma,and decreased the contents of tumor necrosis factor-α(TNF-α)and interleukin 6(IL-6).According to the results of HE staining and oil red O staining of mouse liver pathological sections,it indicated that Sanqidansenshanzha complex prescription could reduce hepatic fat vacuolation and inflammatory infiltration,and decrease the accumulation of fat in the liver tissues of hyperlipidemic mice.The high-glucose and high-fat diet combined with streptozotocin induced diabetes mellitus type 2 mouse model was established to evaluate the in vivo hypoglycemic activity of Sanqidansenshanzha complex prescription.The results showed that the Sanqidansenshanzha complex prescription reduced the liver index in mice,decreased fasting blood glucose(FBG)and insulin(INS)levels in mice,reduced the contents of TC,TG and LDL-C,and increased that of HDL-C in mouse plasma.Further HE staining of mouse liver tissues showed that the Sanqidansenshanzha complex prescription reduced hepatic fat vacuolation,deformation of hepatocytes and restore the morphology of hepatocytes in diabetes mellitus type 2 mice.3.To screen the potential targets and pathways involved in the hypoglycemic and hypolipidemic effects of the Sanqidansenshanzha complex prescription by network pharmacology analysis.20 active components of Sanqidansenshanzha complex prescription were screened through databases and literature.155 common targets of active ingredients and combined hyperlipidemia in type 2 diabetes.Through database and literature screening,20 active components of Sanqidansenshanzha complex prescription were selected.155 common targets of active ingredients and type 2 diabetes with hyperlipidemia were obtained.Through PPI network topology screening,33 core targets were obtained,including MMP1,MMP9,FGF2 and IL10.Go function and KEGG pathway enrichment analysis found that the treatment of diabetes mellitus type 2 combined with hyperlipidemia by Sanqidansenshanzha complex prescription mainly involves lipids and atherosclerosis,PI3 K Akt signaling pathway,NF-κB signaling pathway,insulin signaling pathway,c AMP signaling pathway,and AMPK signaling pathway.4.To obtain the differential metabolites and pathways responsible of the hypolipidemic effects of Sanqidansenshanzha complex prescription based on metabolomics.Through fecal metabolome analysis in mice,a total of 8 differential metabolites were identified in the Sanqidansenshanzha complex prescription group,which were zolpidem,galacto-N-biose,hydroxyisocaproic acid,androstenedione,succinate,7-hydroxy-4-cholesten-3-one,amiloride and prophobilinogen.These metabolites were mainly involved in pathways including primary bile acid biosynthesis,steroid hormone biosynthesis,the citric acid cycle and porphyrin metabolism,in which disorders of bile acid metabolism lead to hyperlipidemia.Through metabolome analysis,it was indicated that Sanqidansenshanzha complex prescription treating hyperlipidemia was associated with bile acid metabolism.5.Sanqidansenshanzha complex prescription exerts its hypoglycemic and hypolipidemic effects by regulating the FXR / FGFR4 / FGF15 pathwayFXR acts as a bile acid receptor,and its FXR / FGF15 / FGFR4 pathway is a key pathway in controlling bile acid synthesis.In this paper,we investigated the mechanism of hypoglycemic and hypolipidemic effects of Sanqidansenshanzha complex prescription based on the FXR/FGFR4/FGF15 pathway.In hyperlipidemia mouse models,after administration of the Sanqidansenshanzha complex prescription,FXR and CYP7A1 m RNA and protein expressions were up-regulated,and FGFR4 m RNA and protein expressions were down-regulated in mouse liver tissues.FXR m RNA and protein were up-regulated,and FGF15 m RNA and protein were down regulated in ileal tissues of mice.In a mouse model of diabetes mellitus type 2,the expression of FXR and CYP7A1 m RNA and protein was up-regulated,and the expression of FGFR4 m RNA and protein was down-regulated in the liver tissues of mice after administration of Sanqidansenshanzha complex prescription.FXR and FGF15 m RNA and protein expression were down-regulated in mouse ileal tissues.The above results indicated that the Sanqidansenshanzha complex prescription exerted its hypolipidemic and hypoglycemic effects by regulating the FXR/FGFR4/FGF15 pathway,up-regulating CYP7A1 expression,promoting bile acid synthesis,and affecting bile acid metabolism. |
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