Integrated Metabolomics And Network Pharmacology To Explain The Mechanisms Of The Herbal Pair Of Salvia Miltiorrhiza And Pueraria Lobata For Treating Type 2 Diabetes

Objective: Type 2 diabetes mellitus(Type 2 diabetes mellitus,T2DM)is a metabolic disease characterized by hyperglycemia.It has a high incidence,a long course of disease and many complications,which brings great pain and economic burden to patients.The effect of Salvia Miltiorrhiza and Pueraria Lobata(DG)drug pair,which was obtained by Mr.Chenyu Zhu who had studied the compatibility of diabetes drugs for many years,has been verified in the early stage of the project,but the mechanism of its action in the treatment of T2 DM remains unclear.Therefore,this study mainly established the T2 DM rat model and treated it with DG.Detection of related biochemical indexes and pathological tissue to evaluate the efficacy of DG in the treatment of T2 DM.The therapeutic mechanism of DG on T2 DM was studied by the combination of network pharmacology and metabonomics,the metabolic markers and related pathways were searched and the related targets were verified to clarify the mechanism of DG in the treatment of T2 DM from the metabolic level and the protein level of computer simulation.Methods: 50 Sprague-Dawley(SD)rats were randomly divided into control group(CON group)and high-fat group.The rat model of type 2diabetes was established by intraperitoneal injection of high-fat diet combined with low-dose streptozotocin(Streptozotocin,STZ)in high-fat group.After the model was established successfully,the high-fat group was randomly divided into model group(MOD group),high and low dose DG group(DGH group,DGL group)and metformin group(MET group).After8 weeks of administration,the serum of rats in each group was collected to detect the biochemical indexes,and the pathological analysis of kidney and pancreas was carried out.The active compounds,key targets and related pathways of DG in T2 DM were screened by network pharmacology.The metabonomics of rat urine were analyzed by LC/MS to screen the differential metabolites and key targets of DG in the treatment of T2 DM,and to analyze the enrichment of metabolic pathway.The interaction between the key targets of network pharmacology and metabonomics was analyzed,and the interaction targets were obtained.The molecular verification was carried out by real-time fluorescence quantitative PCR(RT-q PCR),and the action mechanism of DG on T2 DM was further expounded.Results: Biochemical analysis showed that,compared to the CON group,the content of creatinine(CR),alanine aminotransferase(ALT),total cholesterol(TC),aspartate aminotransferase(AST),triglyceride(TG),low density lipoprotein(L-DLC)and alkaline phosphatase(ALP)in the MOD group increased(P < 0.05).The above indexes were significantly improved after treatment with DG(P < 0.05).Histopathological analysis showed that the pancreas and kidney of T2 DM rats had pathological changes in different degrees,and the pathological changes were improved to some extent after treatment with DG.Metabonomics results showed that compared with the CON group,a total of 147 differential metabolites were significantly changed in the urine of rats in the MOD group.After DG treatment,the changes of 48 metabolites were reversed in varying degrees,and 48 differential metabolites involved 260 targets,mainly involved in 74 metabolic pathways,including amino acid metabolism,c GMP-PKG signal pathway and other pathways.Through the method of network pharmacology,56 active components and 161 T2DM-related targets were screened from DG,which mainly involved 70 metabolic pathways,such as HIF signal pathway,c GMP-PKG signal pathway,insulin signal pathway,and other pathways.The key target proteins of network pharmacology and metabonomics were analyzed by STRING interaction analysis,and a total of 14 interaction target proteins were obtained.14 targets were verified by RT-q PCR,the results showed that T2 DM could cause significant changes in the expression of 11 targets,and the content of 7 targets could be reversed after intervention with DG.Conclusion: DG can improve the changes of biochemical indexes and histopathology in T2 DM rats,regulate the disorder of glucose and lipid metabolism,and protect the function of liver and kidney.The active compounds in DG can cooperate with T2DM-related targets and metabolic pathways to regulate the metabolic disorder in T2 DM rats.This study combined with the methods of network pharmacology and metabonomics to explain the action mechanism of DG in the treatment of T2 DM,and further proved the feasibility and scientificity of this method to study the mechanism of traditional Chinese medicine in the treatment of metabolic diseases.