Inquiry Into The Material Bases Of Early Diabetic Nephropathy With The Pathogenesis As Deficiency Of Both Qi And Yin And Obstruction Of Collaterals Based On The Inflammatory Mechanism By The Sirt1/NF-Kappa B Pathway And Lipid Metabolism Disorder

Objective Diabetic nephropathy(Diabetic Nephropathy,DN)is one of the most main microvascular complications of diabetes.In recent years,more and more researches suggest that DN is an inflammatory disease accompanied by metabolic disorder and the progress of the hemodynamic abnormalities.This study aims to observe the change of inflammatory factors and anti-inflammatory factors before and after the disease process and treatment of DN,which means to explore the material bases of early DN with the pathogenesis as deficiency of both qi and yin and obstruction of collaterals,providing the objective basis for treatment according to syndrome differentiation of DN.Methods The models of earlier diabetic nephropathy in SPF male SD rats(180±20g),were established by fed high fat and sugar diets 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.Rat's condition all the time were observed in 12 weeks.After 12 weeks,the 24h urine was collected to test MAU,Ucr,calculating urinary protein excretion rate and creatinine clearance rate.The blood was collected to test Glu,CHOL,Scr and TXB2,6-keto-PGF1a.The pathological changes from PAS staining of renal tissue were observed.?2-MG in urine was detected by ELISA.By Western-Blot,Sirtl,NF-?Bp65,IL-1?MIF and MCP-1 were monitored to find the relationship between them.By shot lipidomics the majority of lipid species were analysed.Results 1.The diabetic nephropathy rats were in a poor condition.Compared with the normal group,model rats significantly have small size,prominent bones,less muscle.The model group is the most significant.The body weight of model rat was lower than normal group(P<0.01).Compared with the model group,the weight gain(P<0.05)in the high dose Tangshen Prescription group was higher than that in the model group.2.Compared with the normal group,the blood glucose and cholesterol levels in the model group were significantly higher(P<0.05).Compared with the model group,the blood glucose of the traditional Chinese medicine treatment group was decreased(P<0.05).Compared with the model group,the cholesterol level was significantly decreased(P<0.05).Urinary excretion,kidney weight,relative kidney weight,TXA2 of model group were significantly higher than that of the Valsartan group and the high-dose Tangshen Prescription group(P<0.05);The Ccr of the model group was significantly higher than the high-dose Tangshen Prescription group and the Valsartan group(P<0.05).3.Compared with normal group,the blood viscosity,the high shear viscosity of whole blood and low shear whole viscosity of blood in model group were increased(P<0.01)and the moderate shear viscosity of whole blood was increased(P<0.05).Compared with the model group,the blood viscosity,high shear viscosity of whole blood and low shear viscosity of whole blood in each treatment groups were decreased(P<0.01).Moderate shear viscosity of whole blood was decreased(P<0.05).4.The results of PAS staining show that the normal group rats have a clear structure of glomerular,tubular,and glomerular capillary dilation.The increased glomerular,mesangial cell proliferation,mesangial expansion,thickening basement membrane,dilated tubules,epithelial cell vacuoles change were showed in the kidney of model rats.The mesangial cells,mesangial matrix mitigate,the reduced number of tubular dilation and the vacuolation reduction were showed in the cure group.5.Compared with the normal group,the expression of Sirtl in renal tissue of rats was reduced in the model group,and the expression of acetylated NF-kappa Bp65,IL-1,MIF and MCP-1(P<0.01)were increased.Compared with the model group,the expression of Sirtl in eachtreatment groups were increased,and the expression of NF-kappa IL-1,Bp65,MIF andMCP-1 were decreased(P<0.01).Using multiple linear regression analysis,there is a closely relationship between the Sirtl and MCP-1 as well as NF-?Bp65 which has a relationship with MIF?IL-1 and MCP-1.6.Compared with the normal group(630.99+58.91),the total contents of phosphatidylglycerol(PG)in renal tissue were decreased(444.36+42.32,P<0.05),and the PG in the renal tissue with western medicine group also reduced(470.21+78.5,P<0.05).After the treatment of traditional Chinese medicine,these lipids contents,to a large extent,were restored to normal group(603.51+101.4,P>0.05).Compared with the normal group(2.01+).31),the phosphatidylinositol(PI)content of model group showed a decreasing trend(1.69+0.31,P>0.05),in which 16:0-20:4,18:2-20:4,18:0-22:6 PI contents significantly were reduced(P<0.05).After traditional Chinese medicine treatment,16:0-20:4,18:2-20:4,18:0-22:6 PI contents were restored to normal levels(P>0.05).Compared with the normal group(12.29 +4.18),the phosphatidylcholine(PC)of model group was decreased(11.68+3.27,P>0.05),with the clear reduction of the D14:0-16:0?A16:0-18:0?P18;2-20:1 PC content(P<0.05).Conclusions The deficiency of both qi and yin and obstruction of collaterals were main pathogenesis of early DN.Hemodynamic change showed the microcirculation disorder in the DN rats.Glycolipids biochemical and lipid metabonomics analysis showed that the metabolic disorders in the DN rats.The protein expression of Sirtl in the kidney tissue of rats was increased,while the NF-xBp65 expression was decreased.The expression of Sirtl was increased in the Tangshen Prescription group,and the level of acetylation of NF-nuclear factor kappa Bp65 was reduced,showing the pretty relationship that Sirtl can adjust NF-?B p65 and then regulate the produce of the inflammatory.Therefore,anti-inflammatory factor acts as a vital qi role to resist evil in the development of DN.When the inflammation factor continues to be activated,inflammation factors damage vascular cell,leading to the vascular complications.The study shows that the disorder of inflammatory and anti-inflammatory mechanism is the key of the pathogenesis of the deficiency of both qi and yin and obstruction of collaterals in the early DN with the micro pathology basis of the metabolic disorders and microcirculation.