| Objective Establish the rat model of diabetic nehropathy by high fat feed,unilateral nephrectmy and intraperitoneal injection of STZ.Observe the therapapeutic effect of Yiqi Bushen decoction combined with Tipterygium wilfordii on reducing urinary protein in diabetic nephropathy rats,and investigate its molecular mechanism.Methods 60 male SD rats of clean grade,were randomly divided into 6 groups,normal group,model group,western medicine group,Chinese medicine group,Tripterygium wilfordii group,combined treatment group.Except the normal group,the other rats were fed with high fat diet,given right nephrectomy at the end of the 4th week,and intraperitoneal injection of STZ during the 6-8th weekS,to establish the rat model of diabetic nephropathy.During the 9-16th weeks,each treatment groups were given corresponding drugs by gavage.At the end of 0,8,16th weeks,eachrat,urinary albumin to creatinine ratio(UACR)were detected by ELISA.At the end of 8,12,16th weeks,blood glucose level of the rats were detected by Abbott One-Touch glucose Collecting.At the end of 16th week,we collcted the blood,urine of rats,obtained the renal tissue,mearsured the serum creatinine,urea nitrogen and blood glucose level by biochemical analyzer;observed the pathological changes of renal tissue by microscope and electron microscope.The VEGF mRNA expression were measured by RT-PCR,the protein expression by immunohistochenmistyr.By using enzyme-linked immunosorbent determination of rat serum IL-6 production of TNF alpha.Results 1.general situation:after intraperitoneal injection of STZ,the model rats become depressed,activities were reduced,hair light were lost,body weight decreased significantly,the amount of drinking water greatly increased,thin stool,urine volume also increased significantly compared with the previous.In the process of making the model,the there is one rat died of anesthesia in model group.2 Weight:compared to normal rats,the body weight of the rats fed with high fat diet increasea faster(p<0.05);After injection of STZ,the body weight of the model rats were all decreased,compared with the normal group,the difference was statistically significant(p<0.05).3.Indexes:after the intraperitoneal injection of STZ,compared with the normal group,the blood glucose level of the model eas significantly increased t(p<0.01);At the end of the 16th week,compared with the normal group,the blood glucose,urine albumin creatinine rztio,serum creatinine,urea nitrogen were increased,and the expression of VEGF protein and mRNA of the model group were decreased,the difference statistically significant;After drug therapy,compared with the model group,the SCr,BUNof the treatment groups were dereased,the combined treatment group showed best therapeutic effection among the treatment groups;the UACR were decreased.VEGF protein expression and mRNA of the treatment groups were increased,the Tripterygium wilfordii group and the combined treatment group showed besttherapeutic effection among the treatment groups(p<0.05);At the end of 16th,releasing of IL-6 and TNF-αwere increased.After drug therapy,each treatmen is declining,delcline of combined-treatment group is the most obvious among the treatment groups.4.The light microscope showed that compared with the normal group,the glomerular size of the model group increased,mesangial arer widened,capillary pan open healthlessly,renal tubular epithelial cell swelling and vacular degeneration,the glomerular hypertrophy,and mesangial hyperplasia degree of the treatment group reduced compared with model group.The electron microscope showed that the foot process width of the model group increased,the foot process fused and the glomerular basement membrane thickened.All the pathological lesions has been reduced after treatment.This model established by high-fat diet feed and mono-nephrectomy is ideal.Conclusion By high fat diet feed,unilateral nephyrectomy and intraperitoneal injection of STZ,the fasting blood glucose,serum Cr,BUN,and ACR of the model group were significantly increased compared with normal rats;the light and electron microscope showed the number of podocyte in model group decreased,foot process fused and width increased,mesangial broadenes,glomerular basement membrane thickening,thus building Man like rat model of type 2 diabetic nephropathy.Found the yiqibushen decoction and Tripterygium Wilfordii can effectively reduce the fasting blood glucose,urinary albumin excretion,improve renal function and delay the progress of DN.Its mechanism may be related with the down regulating effect on the expression of VEGF and reducing releasing inflammatory factor like IL-6 and TNF-α. |
