Effect And Mechanism Of Huang Di An Xiao Capsules Regulate PI3K/Akt Signaling Pathway On Improving Vascular Disease In Type 2 Diabetes

Objective This subject is based on previous research,GK rat and human umbilical vein endothelial cells are the model,the PI3K/Akt signaling pathway are a starting point,were identified the protective effect and mechanism of Huang Di An Xiao capsules on GK rats with vascular disease in type 2 diabetes to explore.Method1.Random blood glucose >11.1mmol/L was used as the criterion for T2 DM,and the GK rats were randomly divided into 6 groups: model group,metformin group(0.72g/kg),Shen Qi Jiang Tang granule group(1.08 g/kg),Huang Di An Xiao capsules high dose group(12.0 g/kg),Huang Di An Xiao capsules(6.0 g/kg)and Huang Di An Xiao capsules low dose group(3.0 g/kg),the Wistar rats were set up as the normal group,with 10 rats in each group.A model of type 2 diabetic angiopathy was established by adding 0.1mg/mL N?-Nitro-L-argininemethylesterhydrochloride(L-NAME)to drinking water of GK rats combined with high fat diet.The model was given at the same time,1 time /d for 6 weeks.After the experiment,the fasting blood glucose(FBG)of the rats was measured.The systolic and diastolic function of abdominal aorta in GK rats was observed by ultrasonic backscatter.The tail artery blood pressure measuring instrument was used to monitor the systolic pressure(SBP),diastolic pressure(DBP),mean arterial pressure(MAP)and heart rate(HR)in GK rats.The serum biochemical indexes of abdominal aorta(TC,TG,HLDL,LDL,ox-LDL)were detected by automatic biochemical analyzer.Enzyme linked immunosorbent assay was used to detect the contents of ET-1,HIF-1? and VEGF in abdominal aorta.HE,Masson and Verhoeff staining were used to observe histopathological damage degree of abdominal aorta in rats.Immunohistochemical staining was used to observe the distribution of type I collagen(Collagen I)and type III collagen(Collagen III)in abdominal aorta of rats.The expression of PI3 K,PTEN,Akt,HIF-1? and VEGF mRNA in abdominal aorta of GK rats was detected by RT-qPCR.Western blot technique was used to detect the protein expression level of PTEN,p-PI3 K,p-Akt,HIF-1? and VEGF in the abdominal aorta of rats.2.20 Wistar rats were randomly divided into normal group,Huang Di An Xiao capsules group(15.0g/kg),7 rats in the normal group,the Huang Di An Xiao capsules group was13.Huang Di An Xiao capsules group were lavaged with the corresponding dose of drugs,the normal group were given the same amount of solvent,2 times a day for 3consecutive days.The rats were killed after the last administration,the blood was taken from abdominal aorta,the serum was separated,and the serum of same group was combined.30 min was inactivated by water bath at 56 ?,and bacteria removal was filtered by 0.22?m filter,and stored at-70 ?.The effects of different concentrations of Huang Di An Xiao capsules containing serum on proliferation of HUVEC by MTT method,to find out the best concentration of Huang Di An Xiao capsules containing serum.The expression of PI3 K,Akt,HIF-1? and VEGF mRNA in HUVEC was detected by RT-qPCR technique.Western blot technique was used to detect the protein expression level of PTEN,p-PI3 K,p-Akt,HIF-1? and VEGF in the abdominal aorta of rats.Results1.Huang Di An Xiao capsules not only can decrease the blood glucose and lipid metabolism,improve the injury degree of abdominal aorta in GK rats,but also can significantly reduce ET-1,HIF-1?,VEGF,Collagen I and Collagen III expression and PI3K?Akt ?HIF-1??VEGF mRNA and p-PI3K?p-Akt?HIF-1??VEGF protein expression.The expression level of PTEN mRNA and protein was significantly increased(P<0.01 or P<0.05).2.MTT was screened Huang Di An Xiao capsules containing serum optimal dose is25%,the best time is 48 h.Huang Di An Xiao capsules containing serum cansignificantly reduce the expression of HUVECs in PI3 K,Akt,HIF-1?,VEGF mRNA and p-PI3 K,p-Akt,HIF-1?,VEGF protein,significantly increased PTEN mRNA and protein expression level(P<0.01 or P<0.05).ConclusionT2 DM capsule can improve the vascular pathological damage of Huang Di An Xiao capsules,the mechanism may be related to the regulation of the PI3K/Akt signaling pathway and the inhibition of endothelial injury in the abdominal aort.