| Objective: The membranous nephropathy(MN)rat model is established by the method of cationic bovine serum albumin(C-BSA),and Yishen-Qushi-Tongluo medicine is applied to treat it.By observing the indexes such as 24 hours urine protein(24hUTP),serum total protein(TP),albumin(ALB),serum total cholesterol(TC),triglyceride(TG),blood urea nitrogen(BUN),serum creatinine(Scr)and the expression of Nephrin ?NEPH1(mRNA and protein)in the renal tissue,the intervention or therapeutic effects of Yishen-Qushi-Tongluo medicine on the membranous nephropathy and its possible mechanism are explored,so as to provide new ideas and methods for the clinical treatment of membranous nephropathy.Methods: Forty healthy male SD rats weight(160~180g)of clean grade were selected.The rats were fed with common feed for one week.After that the urine of each rat in 24 hours were collected to test 24 hUTP.And the results were all negative.Then the rats were randomly divided into normal group(NG)(n=10)and model group(n = 30).Except the rats in NG,all of the rats in model group were immunized with bovine serum albumin for one week.And then the model rats was injected with C-BSA,while the NG with normal saline for 4 weeks.Then 24 hUTP was tested to make sure that the MN model was established.After that the rats in model group were randomly divided into Chinese medicine group(TCMG),Tripterygium wilfordii + Prednisone acetate group(T+HG)and model group(MG).Rats in T+HG and TCMG were given the corresponding medicine for gavage therapy,while the ones in NG and MG were given the same amount of normal saline for 4 weeks.At the end of the disposition,urine of each rat in 24 hours was collected to test24 hUTP.Then the blood and the renal tissue of each rat was taken away,andthe rats were sacrificed.Automatic biochemical analyzer was used to detect the serum TP,ALB,TC,TG,BUN,Scr.The changes of glomerular basement membrane and rat paw process were observed by light microscope,electron microscope and immunofluorescence.The expression of Nephrin and NEPH1 mRNA and protein in kidney tissue were detected by real-time fluorescence quantitative PCR(real-time PCR)and Western blot.Results:1 24 hUTP test resultsCompared with NG,the other groups showed significant proteinuria with significant difference(P<0.05).In comparison with MG,24 hUTP of the treatment group were significantly decreased(P<0.05).There was no significant difference between the two treatment groups(P>0.05).2 Biochemical indexes of each group2.1Serum TP and ALB testCompared with NG,the serum TP and ALB in other groups were significantly decreased(P<0.05).Compared with MG,serum TP,ALB of the two treatment groups were significantly increased(P<0.05).There was no significant difference between the two treatment groups(P>0.05).2.2 Serum TC and TG testCompared with NG,the serum TC,TG in other groups were significantly increased(P<0.05).Compared with MG,serum TC and TG of the two treatment groups were significantly decreased(P<0.05).In the two treatment groups,serum TC,TG in TCMG was significantly lower than that of T+HG(P<0.05).2.3 Blood BUN,Scr testThere was no significant difference in serum BUN and Scr among every group(P>0.05).3 Pathological observation of renal tissueUnder the light microscope,glomerular structure of NG had no obvious abnormalities.Glomerular epithelial of MG rats had complex red protein deposition.Glomerular basement membrane(GBM)was diffuse thickeningand thPodocyte foot processewas diffuse fusion.The pathological changes of the treatment group was reduced.Under the electron microscope,glomerular structure of NG had no obvious abnormalities.A large number of electron dense deposits arranged orderly on the glomerular subepithelial of MN.Glomerular basement membrane was diffuse thickening and podocyte foot process was diffuse fusion and spike formation.The pathological changes of the treatment group,compared with MG,was reduced and mild thickening of the glomerular basement membrane,electron dense deposits less podocyte foot part of the fusion.Immunofluorescence showed that there was no obvious abnormality in NG,and MG showed that the deposition of IgG along the glomerular capillary wall and part of the mesangial area showed fine granular deposition.Compared with MG,the pathological changes of the two treatment groups were mild,and a small amount of Ig G was found along the glomerular capillary wall.4 Expression of Nephrin and NEPH1 in renal tissue4.1 Real-time PCR resultsCompared with NG,the expression of Nephrin mRNA and NEPH1 mRNA in other groups decreased significantly(P<0.05).Compared with MG,the expression of Nephrin in treatment groups was significantly increased(P<0.05).There was no significant difference between the two treatment groups(P>0.05).4.2 Western Blot resultsCompared with NG,Nephrin and NEPH1 protein expression in other groups were significantly decreased(P<0.05).Compared with MG,the expression of Nephrin and NEPH1 protein in treatment groups was significantly increased(P<0.05).There was no significant difference between the two treatment groups(P>0.05).Conclusions:1 Yis hen-Qushi-Tongluo medicine,like tripterygium wilfordii combinedwith Prednisone acetate can decrease the urinary protein and increase plasma protein of MN rats.2 Yishen-Qushi-Tongluo medicine can improve lipid metabolism and it functions better than tripterygium wilfordii combined with Prednisone acetate.3 Yishen-Qushi-Tongluo medicine can effectively reduce the glomerular basement membrane immune complex deposition in MN rats,thus improving the pathological damage of glomerular basement membrane.4 Yishen-Qushi-Tongluo medicine can increase the expression of Nephrin,NEPH1 mRNA and protein in the renal tissues of MN rats,repair glomerular in the loss of slit diaphragm protein,protect the integrity and permeability of glomerular basement membrane filtration barrier,which may be one of the ways to control proteinuria. |
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