Treatment Of Type 2 Diabetes Mellitus With Jinqi Jiangtang Tablets Through Multiple Target Organs And Multiple Organs

Background: Jinqi Jiangtang Tablet(JQJTT)is a classic prescription for treating diabetes mellitus,which is derived from the classic Chinese herbal medicine "Qianjin prescription" in the Tang Dynasty.It has a variety of components and targets,but the synergistic mechanism needs to be further clarified.Objective: The mechanism of Jinqi Jiangtang Tablets(JQJTT)in treating type 2 diabetes mellitus(T2DM)was studied through multiple targets coordination with multiple organs.Methods:1)Network enrichment: the components of Astragalus membranaceus,Coptis chinensis and honeysuckle were searched through TCMSP,TCMID database and literature;TCMSP,Swiss Target Prediction,SEA and Pharmmapper were used to screen targets related to components,and the biological functions of related targets were classified through Matescape;The genes related to T2 DM were screened by Genecards and OMIM database,and the intersection genes were obtained by intersecting these genes with prediction targets of JQJTT;Using the Clusterprofiler R package in R,the tissue and organ enrichment of intersection genes was analyzed(p < 0.05).According to the tissue and organ classification of cells in the enrichment results,the organs and tissues closely related to T2 DM were found and analyzed respectively;Using KEGG database,the targets enriched in various organs were enriched,and the relevant pathways and targets were screened.Finally,the hub genes were screened by PPI analysis combined with literature retrieval.2)Cell experiment verification: Hep G2 and C2C12 cells were stimulated by high concentration of glucose to become the model of insulin resistance caused by glucose toxicity;The glucose intake of cells stimulated by insulin was detected to evaluate insulin sensitivity;Real time quantitative polymerase chain reaction(RT q PCR)and Western blotting(WB)were used to analyze the expression changes of related targets affecting insulin signaling pathway in hepatocytes and myotube cells,verify the mechanism of JQJTT treating T2 DM through liver and skeletal muscle,and verify the synergy analyzed by system pharmacology.3)Animal experiment verification: Male Kunming mice were fed with high-fat diet(HFD)and injected with streptozotocin(STZ).After T2 DM model was established,they were treated with JQJTT.The changes of body weight and fasting blood glucose were observed and recorded.After treatment,oral glucose tolerance test(OGTT)and insulin tolerance test(ITT)were carried out;After the mice were killed under anesthesia,the serum of mice was collected to detect four items of blood lipid and two items of liver function including aspartate aminotransferase and alanine aminotransferase;The adipose tissue sections of mouse liver,pancreas and epididymis were collected for staining,the pathological phenomena were observed,and the corresponding staining results were counted and analyzed.Results:1)Through various databases and literature retrieval,441 chemical components were collected,with 2655 prediction targets and 9352 genes related to T2 DM.After the intersection of the two,1346 genes were obtained;After organ enrichment analysis,the intersection genes were enriched in liver,skeletal muscle,pancreas and adipose tissue;The KEGG pathway enriches the predicted targets in the liver and skeletal muscle.JQJTT in the liver can treat T2 DM by regulating bile acid secretion,apoptosis and circadian rhythm,respectively.In skeletal muscle,JQJTT cooperatively treats T2 DM by regulating PI3K/AKT signal pathway,MAPK signal pathway,Rap1 signal pathway,RAS signal pathway,EGFR tyrosine kinase inhibitor resistance and apoptosis.2)Through Hep G2 cell experiment,JQJTT can alleviate insulin resistance caused by high glucose in hepatocytes.At the gene level,JQJTT up-regulated the expression of ABCC2 and AKT and inhibited the expression of GSK3-β、 FOXO1 and TNF-α;At the protein level,JQJTT can activate the PI3K/AKT pathway in the liver by increasing the secretion of bile acids,and then inhibit GSK3-β,and promote glycogen synthesis by PI3K/AKT/GSK3-β pathway;JQJTT can also inhibit the expression of FOXO1,thus inhibiting the expression of gluconeogenic enzyme,and inhibit gluconeogenesis through PI3K/AKT/FOXO1 pathway.Therefore,JQJTT can reduce hepatic glucose production to improve T2 DM,promote glycogen synthesis and inhibit gluconeogenesis,so as to reduce blood glucose and improve T2 DM.3)Through C2C12 cell experiment,JQJTT can also alleviate insulin resistance caused by high glucose in myotube cells.At the gene level,the results show that JQJTT reduces CASP3 and IKKB and inhibits apoptosis and inflammation induced by MAPK pathway;JQJTT also up-regulated the expression of ITGB1,PI3 K and GLUT4,which proved that JQJTT can promote insulin signal,increase the expression and translocation of GLUT4 in skeletal muscle.Therefore,JQJTT can promote glucose transport,reduce peripheral blood glucose,chronic inflammation and apoptosis,and improve hepatic insulin resistance.4)Through animal experiments,JQJTT can prevent the continuous rise of blood glucose caused by HFD and STZ,and even reduce blood glucose;Through OGTT and ITT observation,JQJTT can significantly improve systemic blood glucose homeostasis and insulin sensitivity;The results of four blood lipid tests showed that JQJTT could effectively reduce blood lipid and increase the content of high density lipoprotein(HDL-C);In the results of tissue staining sections,the results of HE staining of liver showed that JQJTT could improve cytoplasmic looseness,steatosis and inflammation of liver.The results of HE staining of adipose tissue and pancreas showed that JQJTT could alleviate inflammation and erythrocyte sedimentation,coordinate glucose and lipid metabolism and treat T2 DM.Conclusion:1)The network enriched and sorted out the components and corresponding prediction targets of three traditional Chinese medicines of JQJTT,then screened out the common targets of prediction targets in T2 DM related genes,and screened out four tissues and organs closely related to T2 DM,namely liver,pancreatic islet,adipose tissue and skeletal muscle.This enrichment result preliminarily proves that the traditional Chinese medicine compound plays a synergistic role in the treatment of diseases through multiple targets and multiple tissues and organs.At the same time,it also makes basic preparations for the next research.2)Cell experiments found that JQJTT can pass through the bile acid secretion pathway in the liver,activate the insulin signal pathway through bile acid,promote glycogen synthesis and inhibit gluconeogenesis,so as to reduce blood glucose and improve T2 DM.It can inhibit skeletal muscle inflammation and apoptosis,promote insulin signal and increase peripheral glucose intake and consumption.3)JQJTT can improve systemic blood glucose homeostasis and insulin sensitivity,hepatic steatosis and inflammation,as well as adipose tissue and pancreatic inflammation.Therefore,JQJTT can improve the disorder of glucose and lipid metabolism through multiple organs.