| ObjectiveHuang Di An Xiao capsule is the First Affiliated Hospital of Anhui University of Chinese medicine in treatment of type 2 diabetes mellitus(T2DM)characteristics of hospital preparations.Prescription from Loquat leaf,Pueraria,Ophiopogon,Rehmannia glutinosa and other six herbs,clinical application for many years,the exact effect,but its impact on the hepatic insulin signaling pathway is unclear.The aim of this study was to investigate the regulatory effect of Huang Di An Xiao capsule on insulin IRS-1/PI3K/Akt/GSK-3? signaling pathway and its molecular mechanism of lowering blood glucose.Method 1 Effects of Huang Di An Xiao capsule on IRS-1/PI3K/Akt/GSK-3? signal pathway in spontaneous type 2 diabetic model of GK ratsBlood glucose was measured after tail vein disinfection in GK rats,with the random blood glucose more than 11.1mmol/L as a criterion for T2 DM model.GK rats were selected and randomly divided into model group,Shen Qi Jiang Tang cranule(1.08 g/kg)and Huang Di An Xiao capsule(12.0?6.0?3.0 g/kg),ten Wistar rats served as control group.After 6 weeks of administration,fasting blood glucose(FBG),random blood glucose(RBG),serum insulin(FINS),fasting serum glucose(FPG)and insulin resistance index(HOMA-IR),oral glucose tolerance test(OGTT)were performed.The levels of(TC,TG,LDL-C),glycosylated hemoglobin(HbA1c),glucagon like peptide-1(GLP-1)and insulin like growth factor(IGF-1)were measured.The pathological changes of pancreas and liver were observed by hematoxylin-eosin staining(HE).The synthesis of hepatic glycogen was observed by PAS staining.The mRNA expression of INSR,IRS-1,PI3 K,Akt,GSK-3? and GS were detected by RT-qPCR.The expressions of INSR,p-IRS-1,PI3 K,p-Akt,p-GSK-3? and GS were detected by Western blot.The expression of p-IRS-1,PI3 K,p-Akt and p-GSK-3? were detected by immunofluorescence staining.2 Effects of Huang Di An Xiao capsule containing serum on IRS-1/PI3K/Akt/GSK-3? signal pathway induced by high glucose and insulin in HepG2 cellsHepG2 cell culture medium at 5% CO2 with DMEM medium,37? in incubator,using high glucose and insulin induced HepG2 cells replication insulin resistance model,MTT method for screening Huang Di An Xiao capsule best serum concentration and action time.HepG2 cells were divided into normal control group,high glucose and insulin stimulation group,high glucose and insulin stimulation+drug-containing serum group.The expressions of INSR,IRS-1,PI3 K,Akt,GSK-3? and GS mRNA were detected by RT-qPCR and the expressions of INSR,p-IRS-1,PI3 K,p-Akt,p-GSK-3? and GS were detected by Western blot.The expression of INSR,p-IRS-1,PI3 K,p-Akt,p-GSK-3? and GS protein were detected by immunofluorescence technique.Results 1 Effects of Huang Di An Xiao capsule on IRS-1/PI3K/Akt/GSK-3? signal pathway in spontaneously type 2 diabetic model of GK ratsThe levels of FBG,FPG,RBG,HOMA-IR,TG,TC,LDL-C and AUC of GK rats in Huang Di An Xiao capsule high and medium dose group were significantly decreased;Pancreas and liver tissue structure changes significantly reduced.At the same time,up-regulation of INSR,IRS-1,PI3 K,Akt and GS mRNA and downregulation of GSK-3? mRNA,increased expression of INSR,p-IRS-1,PI3 K,p-Akt and GS proteins,decreased p-GSK-3? protein expression.2 Effects of Huang Di An Xiao capsule containing serum on IRS-1/PI3K/Akt/GSK-3? signaling pathway induced by high glucose and insulin in HepG2 cellsThe MTT assay screened 10% drug serum,the role of 48 h,higher cell activity.At this time,Huang Di An Xiao capsule could significantly up-regulate INSR,IRS-1,PI3 K,Akt and GS mRNA,down-regulate GSK-3? mRNA,increasing INSR,p-IRS-1,PI3 K,p-Akt and GS protein expression,reducing p-GSK-3? protein expression.Conclusion 1 The Effects of Huang Di An Xiao capsule in regulating blood glucose,insulin resistance and IRS-1/PI3K/Akt/GSK-3? signal pathway in diabetic GK rats 2 The Huang Di An Xiao capsule regulates IRS-1/PI3K/Akt/GSK-3? signal pathway protein activity,reduced the content of glucose and increased glycogen synthesis,which may be one of the mechanisms of improving insulin resistance... |
PDF Full Text Request