The Effect And Mechanism Of Tangshen Recipe On Inflammation In Rats With Diabetic Nephropathy Were Studied From The Sirt1/NF-?B Pathway

Objective The study intends to observe the influence of Sirt1?acetylation NF-KBp65 as well as inflammatory cell infiltration on diabetic nephropathy rats based on verify the Tangshen prescription.We try to investigate the anti-inflammatory effects of Tangshen prescription on kidney tissue and the mechanism from the Sirt1-NF-?Bp65 signaling pathway.Methods The earlier model of diabetic nephropathy in SPF male SD rats(180±20g),was established by fed High fat and high sugar diet and intravenously injection of STZ(30mg/kg).They were divided randomly into normal group,model group,low dose Tangshen Prescription group,moderate dose Tangshen Prescription group,high dose Tangshen Prescription group,and valsartan group.And intervene them for 12 weeks.Rat's condition all the time were observed;Fasting blood glucose,blood rheology,blood and urine creatinine were detected by automatic analyzer.?2-MG in urine were detected by ELISA.Pathological changes in renal tissue were observed by PAS dying.By Western-Blot,to monitor Sirt1,NF-?Bp65,IL-1?MIF and MCP-1 as well as find the relationship between them.By immunohistochemistry to monitor macrophagesResults(1)The diabetic nephropathy rats were in a poor condition and lost weight.Compared with the normal group,model group lost weight P<0.01.Compared with the model group,the high dose of Tangshen prescription group got fatter,P<0.05.Compared with the normal group,the model group rats in a high blood sugar condition in limosis,P<0.01.Compared with the model group,low dose Tangshen Prescription group,moderate dose Tangshen Prescription group decrease in blood sugar,P<0.05;in blood sugar of high dose of Tangshen prescription group decrease apparently.P<0.01.Compared with the normal group,the model rats increase in?2-MG(p<0.01.Compared with the model rats,the cure group rats decrease in?2-MG(p<0.01).Compared with the normal group,the cure group rats decrease in creatinine clearance(p<0.05),compared with the model rats,high dose of Tangshen prescription group and valsartan rats increase in creatinine clearance(p<0.05).(2)Hemorheology:Compared with normal group,model group blood viscosity,high shear whole blood viscosity and low shear whole blood viscosity increased,P<0.01.Moderate shear Whole blood reduced viscosity increased,P<0.05.Compared with the model group,each treatment group blood viscosity,high shear whole blood viscosity and low shear whole blood viscosity decreased,P<0.01.Moderate shear Whole blood reduced viscosity decreased,P<0.05.(3)Pathology:the results of PAS staining show the normal group rats have a clear structure of glomerular,tubular,and glomerular capillary dilation.the model rats kidney show increased glomerular,mesangial cell proliferation,mesangial expansion,basement membrane thickening,Dilated tubules,epithelial cell vacuoles change.The cure group rats show mesangial cells and mesangial matrix mitigate,reduce the number of tubular dilation,vacuolation reduction,the immunity group rats show that glomeruli,renal interstitial macrophage infiltration and the cure group rats significantly reduced macrophage infiltration.(4)Western-Blot:Compared with the normal group,the rats kidney of model group Sirtl decrease NF-?Bp65?IL-1?MIF and MCP-1 increase,and P<0.01.the cure group increase in Sirtl,NF-?Bp65?IL-1?MIF and MCP-1 decrease,compared with the model group,P<0.01.Using multiple linear regression analysis,there is a closely relationship between the Sirtl and MCP-1 as well as NF-?Bp65,the NF-?Bp65 have a relationship with MIF?IL-1 and MCP-1.Conclusion First,Tangshen Prescription can decrease the rats Kidney damage,improve renal function,and protect the diabetes rats kidney.Second,diabetic nephropathy rats hemorheological change which indicate diabetes rats were in a circulatory disorders,the same as the medicine pathogenesis Tangshen Prescription can ameliorate the index of hemorheology,and further revealed pathogenesis of diabetic nephropathy.Third,the kidney tissue inflammatory cytokines of the diabetic nephropathy rats increased and macrophage infiltration,while Tangshen Prescription can suppress them,which prompt inflammatory play an important role in the rats kidney damage,so Tangshen Prescription can protect the diabetic kidney by anti-inflammatory.Last,the Kidney tissue of diabetic nephropathy rats increased in Sirtl protein,decreased in NF-?Bp65,while Tangshen Prescription can increase both of them and have a pretty relationship,which indicate Sirt1 can adjust NF-?B p65 and then regulate the produce of the inflammatory.