The Protective Effect And Mechanism Of Wanjinwenwu Decoction On Diabetic Cardiomyopathy In Rats

Objective1.To observe the effect of WanJinWenWu decoction on serum index and morphological structure of diabetic rats with cardiomyopathy.2.To elucidate the protective mechanism of WanJinWenWu decoction on diabetic rats against cardiomyopathy;3.To discuss the mechanism of action of WanJinWenWu decoction on diabetic rats against myocardial ischemia/reperfusion injury.Methods1.Totally 60 healthy male SD rats:including Control group with 10 normal rats,and the other group of remaining 50 rats with intraperitoneal injection of 65 mg/kg streptozotocin(STZ).72 h later tested the fasting blood glucose ?11.1 mmol/L and determined the diabetic models were successful made.After 1 month's normal feeding,provide dintragastric administration.2.Diabetic rats were randomly divided into 5 groups:(1)diabetes mellitus group(DM group)with administration of 1 mL/kg/d normal saline;(2)WJL decoction group(WJL group)with administration of 300 mg/kg/d;(3)WJH decoction group(WJH group)with administration of 600 mg/kg/d;(4)metformin group(Met group)with administration of 140mg/kg/d;(5)ALT group(ALT group)with administration of200 mg/kg/d.For 8 weeks,once a day.3.After 8 weeks' intragastric administration,5 rats from each group received anesthesia,and left anterior descending coronary artery for construction of myocardial ischemia/reperfusion model(ischemia 30min/reperfusion 60 min).While the normal Sham Control group performed without ligation.Continuously observed the changes of electrocardiogram.4.Test the changes of serum TC,TG,LDL-C,HDL-C,SOD,MDA,CK-MB and GHb in blood were detected by Elisa kit.Adopted Western Blot to test the protein expression of myocardial cells HIF-1?,HO-1,VEGF,NF-?B,GSK3p,p-GSK3?,Bax,Bcl-2,Caspase-3,and Cleaved-Caspase-3.Observed through electron microscope ultrastructure of myocardial tissue.Detected changes of HIF-1? and VEGF by immunohistochemistry test.Observed changes of myocardial cell structure upon HE staining.Result1.After 8 weeks' oral administration,compared WJH group with DW group,,the blood glucose and GHb decreased significantly(P<0.05),the serum TC,TG,LDL-C,MDA and CK-MB decreased significantly(P<0.01),and the value of serum SOD and HDL-C increased significantly(P<0.01).2.Western Blot method showed that with regard of diabetic cardiomyopathy,compared with DM group,the level of HIF-1? and HO-1 decreased significantly(P<0.05),the level of inflammatory factor(NF-?B)decreased significantly(P<0.05),the level of p-GSK3?/GSK3? increased significantly(P<0.05),and the level of Bcl-2/Bax increased significantly(P<0.05),the level of Cleaved-caspase-3/caspase-3 increased significantly(P<0.05).With regard of myocardial ischemia/reperfusion injury,compared with DM+ I/R group,the level of HIF-1? and HO-1,VEGF of WJH+I/R group increased significantly(P<0.05),the level of inflammatory factor NF-?B decreased significantly(P<0.05),the level of p-GSK3?/GSK3? increased significantly(P<0.05),the level of Bcl-2/Bax increased significantly(P<0.05),and the level of Cleaved-caspase-3/caspase-3 increased significantly(P<0.05).3.The results of electron microscope showed that,compared with the DM groups,there were different levels of improvement in-both WJH group and WJL group.4.The results of immunohistochemistry test,compared with the DM groups,the expression of HIF-la and VEGF in WJH group decreased.significantly(P<0.05).5.HE staining showed there were improvement and morphologic changes of myocardial alignment caused by diabetic cardiomyopathy in both WJH group and WJL group.Conclusion1.WanJinWenWu decoction played a protective role in diabetic cardiomyopathy injury in rats.The mechanism might be related to regulating HIF-1?,further regulating the expression of HO-1 and VEGF at the backward position,inhibiting inflammatory factors such as NF-?B,and lowering the activity of phosphorylated GSK-3?,decreasing Bcl-2/Bax ratio and the expression of apoptosis-related proteins such as caspase-3.2.WanJinWenWu decoction could improve myocardial ischemia/reperfusion injury in diabetic cardiomyopathy rats.The mechanism migth be related to regulating and controlling HIF-1?/HO-1 and HIF-1?/VEGF signaling pathway,reducing the expression of NF-?B and other inflammatory factors,and inhibiting the activity of phosphorylated GSK-3?,the down-regulation of Bcl-2/Bax ratio,and the expression of apoptosis-related proteins such as caspase-3.