The Protective Effect Of Yishen Tongluo Decoction On Renal Tissue Of Rats With Membranous Nephropathy And Effect In The Expression Of Renal Nephrin MRNA And Podocin MRNA

Objective: This research builds Membranous Nephropathy(MN) modelof rats by Cationic Bovine Serum Albumin(C-BSA),and applies YishenTongluo decoction to the model for interventional treatment, though theobservation of biochemical factors of rats (such as24-hour urine protein,serum total protein, albumin, blood fat, creatinine, urea nitrogen and so on)andthe expression of renal Nephrin、Podocin, discussing the effect and probablemechanism of Yishen Tongluo decoction on MN, in order to provideexperimental and theoretical basis for clinical treatment of MN.Methods:60healthy male Wistar rats were randomly divided into2groups,as follows, normal group(NG) of15,model group(MG) of45.ExceptNG, after1week of the pre-immunization the model group once every otherday injected C-BSA via caudal vein to copy MN rats model. The24-hoururine protein were tested after continuous injection for4weeks. Afterconfirming the success of modeling, the rats of model group were divided into3groups,as floolws,model group(MG),Benazepril treatment group(BG),andYishen Tongluo decoction treatment group(TCMG).The rats of BG weregiven Benazepril at a dosage of10mg/kg/d by gavage administration. The ratsof ZG were given Yishen Tongluo decoction by gavage at a dosage of3ml/d.The rats of NG and MG were fed the same dose of normal saline. All ofthe4groups continued dosing for4weeks. After that,24-hour urine and24-hour urine protein of all rats which were keeping abrosia and freelydrinking was determined. And then, putting all rats to death, serum totalprotein(TP), albumin(ALB),total cholesterol (TC),triglyceride(TG),ureanitrogen(BUN), creatinine(Scr) were assayed. Pathological changes of renaltissues of rats were observed by immunofluorescence, light microscopy and electron microscopy. The expression of Nephrin mRNA and Podocin mRNAin the renal tissues were tested by Real-time PCR.Results:1General condition of experimental ratsThe rats of NG were normal diet and urine volume, bright eyes, promptresponse, and plump muscle. After injecting C-BSA via caudal vein, the ratsof MG all showed obvious anorexia, lassitude, baldness, and less activities.Some of them appeared ascites and scrotal edema. Comparing with NG, theother3groups rats were distinctly much thinner, smaller, and baldness in thelast week of experiment. The MG was the most obvious. During the modeling,the rats of model group had three rats death. The cause of death might beanaphylactic reaction of C-BSA and serious infection, which led to organfailure.2Comparison of24-hour urinary proteinAfter4weeks’ injection of C-BSA, the24-hour urinary protein of MGwere rised obvious than these of NG(P<0.05);after4weeks’ administration,24-hour urinary protein of BG and ZG were declined than MG(P<0.05),butthere was no difference between BG and ZG(P>0.05).3Blood biochemical indexCompared with NG, the TP and ALB were declined markedly, and theTC and TG were increased significantly in MG,BG and ZG. There weresignificant differences among them(P<0.05).Compared with MG,the TP andALB were increased markedly, and the TC and TG were declined. They hadsignificant differences(P<0.05).There were unconspicuous differences and hadno statistical significance between ZG and BG(P>0.05).BUN and Scr of allgroups had no statistical significance(P>0.05).4Pathomorphism of renal tissuesThe results of immunofluorescence: There had no evident changes in NG.Diffuse granular IgG and C3deposited along glomerular capillary wall inMG.ZG and BG showed a little IgG and C3deposition.The results of light microscopy: NG have no obviously abnormal changes.MG showed that the basement membrane of glomerular capillaryloops had thickened evidently, the endothelial cell nucleus increasedsignificantly, and rete pegs became larger. Comparing with model group, thethickened basement membrane of capillary loops in BG and ZG relativelyrelieved, the endothelial cell nucleus increased slightly, a small amount of retepegs had formed.The results of electron microscopy: The rats of NG had a clear structureof podocyte and aligned foot processes.MG showed glomerular capillarybasement membrane thickened obviously, amount of similar electron-densematerial deposited under the glomerular epithelium, foot processes fused, andrete pegs formed. The glomerular capillary basement membrane thickenedslightly in ZG and BG. There were a little electron-dense material depositedunder the glomerular epithelium.Part of podocyte foot processes began to fuse.5The expression of renal Nephrin mRNA、Podocin mRNAStatistical analysis showed: compared with NG, Nephrin mRNA、PodocinmRNA of MG had a low level expression, and had significantdifferences(P<0.05).Compared with MG, the expression of Nephrin mRNA、Podocin mRNA of ZG and BG made something increase, and had markedstatistical differences(P<0.05).There were no obvious differences between thetwo treatment groups(P>0.05).Conclusions:1Yishen Tongluo decoction can decline urine protein, increase serumalbumin,improve blood fat of MN rats.2Yishen Tongluo decoction can inhibit the deposition of immunecomplex in membranous nephropathy glomerular basement membrane andthickening of basement membrane of rats,relieve renal pathological injury,anddelay the further grogress of membranous nephropathy.3Yishen Tongluo decoction can revise the expression of NephrinmRNA、Podocin mRNA in renal tissue of membranous nephropathy ratsupwards.The therapeutic action is probably relative to protectingpodocyte,maintaining the integrality of podocyte slit membrane structure and the glomerular filtration function.