The Efficacy Research Of Immune Inhibitors Sequential Therapy According To The Cell Cycle For The GN

Objective:1.To establish the model of Rat AIN similar to the hunman Renal glomerular disease in patho and amynology.2.To assess the therapeutic effects of different sequential therapy regimens according to the theory of cell cycle for adriamycin-induced nephropathy(AIN) rats.3.To compare the effects of sequential regimens and combined regimens by CsA and MMF for AIN rats.4.To establish the schema of clinical medication and collect clinical case. Methods:1.Rat AIN- FSGS model was established by vena caudalis introduction. The level of 24 hour urinary protein, serum total protein(TP),albumin(ALB), cholesterin(Chol),triglyeride (TG),serum urea nitrogen(BUN),serum creatinine(Scr)were measured.The renal pathological changes were observed by light microscope and electron microscope.The expression of FN,LN and CTGF were analyzed by immunohistochemistry. MMP-2,MMP-9 and TGF-β₁ were detected by ELISA. 2.SD rats were randomly divided into five groups: control group (n=8), AIN model group (n=8), MP+CsA+MMF treated group (n=8),MP+CsA+CTX treated group(n=8) and MP+FK506+Rapa treated group (n=8).The level of 24 hour urinary protein, serum total protein(TP), albumin(ALB), cholesterin(Chol),triglyeride(TG),serum urea nitrogen (BUN),serum creatinine (Scr)were measured. The renal pathological changes were observed by light microscope.The expression of nephrin and podocin were analyzed by immunohistochemistry and immunofluorescence(IMF),westernblot was used to detect the experession of CTGF, The expression of nephrin,podocin and CD2APmRNA were analyzed by RT-PCR.MMP-2,MMP-9,TGF-β₁ in serum were detected by ELISA. 3.SD rats were randomly divided into four groups: control group (n=8), AIN model group (n=8) and MP+CsA+MMF sequential therapy group (n=8) and MP+CsA+MMF combination group (n=8).The level of 24 hour urinary protein, TP, ALB,Chol,TG,BUN,Scr, alanine aminotransferase(ALT) and aspartate aminotransferase(AST) were measured;observe the pathological change of liver and nephridial tissue.4.Collect the case of primary glomerulopathy,assess the effect of sequential therapy for the patents who are experiencing hormonal resistance or hormone dependent, Urine protein and biochemical indicator were dedected.Results:1.Compared with control group,the level of 24 hour urinary protein, TG and Chol in AIN model group were obviously increased in treatment groups at 2th,4th,8th and 12th week(P<0.01),TP and ALB were obviously decreased;FSGS were observed in 12th week group; The level of TGF-β₁ in serum were obviously increased,The level of MMP-2 and MMP-9 reduced by degrees;the expression and distribution of FN,LN and CTGF increased significantly.2.Three kinds of sequential combined regimens can reduce the Urine protein in Adriamycin-induced nephropathy rats. Compared with control group,the level of 24 hour urinary protein in AIN model group was obviously increased in treatment groups at 2th,4th,8th and 12th week(P <0.01), the level of 24 hour urinary protein was obviously decreased in treatment groups at 8th and 12th week than in AIN model group(P <0.01); The level of TP and ALB was significant lower in AIN model group than in control group(p<0.01),the level of TG and Chol were significant higher in AIN model group than in control group (P <0.01); The level of TP and ALB was significant higher and the level of TG and Chol were significant lower in treatment groups than in AIN model group. The results of immunohistochemistry indicated the expression of nephrin,podocin and CD2AP in AIN model group rats decreased at 12th week(P <0.01) , the expression of them in treatment groups rats are obviously higher than those in AIN model group(P <0.01). The results of westernblot indicated the expression of CTGF in AIN model group rats increased at 12th week(P <0.01) , the expression of CTGF in treatment groups rats are higher than those in AIN model group(P <0.05); The effect of inhibitting fibrous degeneration in MP+FK506+Rapa treated group are more effective then other treated group. 3.Two kinds of regimens by CsA and MMF can improve the pathological changes in Adriamycin-induced nephropathy rats , raise the level of TP and ALB and reduce the level of TG and Chol in blood serum;but the combined regimens can lead to hepatotoxicity.Conclusions:1.FSGS were observed at 12th week in model group.2.Sequential combined regimens according to the cell cycle can improve the pathological change in Adriamycin-induced nephropathy rats,reduce the Urine protein,increase the level of TP and ALB,decrease the level of TG and Chol;It also can increase the expression of nephrin,podocin and CD2AP ,ameliorate kidney fibrosist. 3.The sequential regimen by CsA and MMF can avoid adverse effect,safer than the combined regimen.4.The clinical therapeutic effects of sequential therapy regimens according to the theory of cell cycle are significant,but still need more clinical authentication.