| Systemic lupus erythematosus(SLE) is a autoimmune disease which severitly harm tohuman healthy. Kidney is the most sensitive organ which can be involved in this disease andnamed with lupus nephritis(LN).At present , the mechanism of SLE is not clear.Exploring themechanism and therapeutic methodes from the direction of Tranditional Chinese Medicine andbinding Western Medicine has been one important pathway to investigate the disease. This topicrefer to the methods of Bergjk and induce successfully the lupus nephritis murine models ofchronic graft-versus-host disease(cGVHD),At the same time, Observing the effects ofBushenQingredu Recipe(BQR)on the models. BQR has been clinically confirmed to havebetter effects on the patients with LN.Additionally ,the topic investigae the pathologicalmechanisms of LN by apoptosis in kidney and the disequilibrium of Th1/Th2 in cGVHD mice. The topic include four parts: 1. Induction of lupus nephritis models of chronic graft-versus-host disease To induce murine lupus nephritis models,we select 6-8 weeks (DBA/2×C57BL/6J)F1hybrids mice and inject parental DBA/2 mice lymphoid cells by caudal vein.Spleen,thymiandlymph nodes(inguinal,axillary,cervical and mesenteric)were removed from female donor miceunder aseptic condition .Single-cell suspensions were prepared with sterile 0.9% NaClsolution(NS).At days 0,3,7and 10, female recipients were injected intravenously(iv)with 50×106viable lymphocytes in 0.3ml NS..At weeks 2,4,8,10and12 after first injection oflyphocytes,urines were collected from all experimental mice.Albuminuria were determined ByCoomassie brilliant blue methods.At the same time ,6 mice were sacrificed to detectedantibody,serum creatinine,blood urea nitrogen and pathological damage of kidneytissue.Antibody(dsDNA) were detected by indirect immunofluoresence,pathological damage ofkidney tissue were observed by the staining PAS,Masson,PAM and electron microscopy.Theresults diplay that two week after injection of lymphocytes,antibody and proteinuria wereproduced ,serum creatinine and blood urea nitrogenthe increased after four week,glomerularmesangial cells increased significantly,interstitial without damaging .At eight week,All theindexes changed significantly,the pathological damage of kidney tissue was glomerularmesangial cells diffuse increased,protein casts appeard .From ten to twelve week,glomerularappeared focal or diffuse hyalinosis and glomeorulosclerosis . immunofluorescence showeddeposits of immunoglobulins IgG,IgM,C3 were observed along the glomerular capillary andmesangial。All the results suggested that the murine models is comparable to the lesionscharacterizing human LN and offer an excellent model for studies on LN. 2. Effects of BQR on murine lupus nephritis models of chronic graft-versus-hostdisease 4 weeks after first injection of lymphocytes,34 female murine were randomly divided intofour groups ,BQR group(10),prednisone group(10),model group(6) and control group(6).8 weeks later,all the mice were killed and observed the index as part one. The results diplay thatthe levels of albuminuria,antibody,serum creatinine and blood urea nitrogen were decreased,albumin were increased in both BQR group and prednisone group. In addition,BQR coulddecrease cholesterol.BQR group and prednisone group could decrease glomerular mesangialcell and infiltrating leucocyte . This result suggested that BQR has better therapeutic effects oncGVHD mice. 3. Effects of BQR on apoptosis in kidney of lupus nephritis models 32 female murine were randomly divided into 4 groups as part two .Immunohistochemistryand western blot was used to detect the protein level of Fas,FasL.the transcription of Fas,FasLmRNA in kidney were detected by RT-PCR, apoptosis was observed by TUNEL assay. Theresults diplay that apoptosis in kidney increased in BQR group and prednisone group, theprotein and mRNA level of Fas and FasL increased in BQR group and predison group. Thissuggested that BQ... |
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