Col4α3NC1中多肽序列pCol(28-40)诱导抗肾小球基底膜肾炎大鼠模型的建立

Establishment of anti-glomerular basement membrane nephritis in rats induced by peptide sequence of CoI4α3NC1

目的为研究人类抗肾小球基底膜肾炎(anti-glomerular basement membrane nephritis,anti-GBM)建立大鼠自身诱导的抗-GBM肾炎模型。方法健康SD大鼠40只,随机分为2组:抗-GBM肾炎模型组和正常对照组。将合成的大鼠肾小球基底膜Ⅳ型胶原α3链非胶原1区(Noncollagen Domain 1 of alpha 3 chain of typeⅣCollagen,Col4α3NC1)中的多肽序列—pCol(28-40)为抗原多肽,皮下注射免疫抗-GBM肾炎模型组;正常对照组同部位注射PBS。于第7、14、21、28、35天检测大鼠24 h尿蛋白、血肌酐及血尿素氮等生化指标变化,普通光镜观察HE,PAS染色肾组织病理改变;免疫荧光观察IgG的沉积;电镜观察肾小球基底膜,足突细胞,免疫复合物沉积的变化。结果大鼠免疫后,抗-GBM肾炎模型组在各时相点的24 h尿蛋白量、血肌酐、血尿素氮明显高于对照组(P<0.01);肾组织病理表现为肾小球内细胞数明显增多,大量细胞性新月体形成,蛋白管型多见,内皮细胞脱落、坏死;免疫荧光检查IgG沿肾小球弥漫性线样沉积;电镜观察肾小球基底膜不规则增厚,足突融合;正常对照组均未见改变。结论成功的建立了大鼠抗-GBM肾炎,该模型病变与人类抗-GBM肾炎的病变较为一致,可用于探讨抗-GBM肾炎的实验研究。

We aimed to establish a rat model of the self-induced anti-glomerular basement membrane（anti-GBM） nephritis for the research of human anti-GBM nephritis pathogenesis.Forty healthy SD rats were randomly divided into 2 groups（the glomeru-lonephritis group and the normal control group）.The anti-GBM nephritis group was injected with the pCol（28-40） antigen peptide of noncollagen domain 1 of alpha 3 chain of typeⅣcollagen（Col4α3NC1） of rat glomerular basement membrane;the normal control group was injected with PBS at the same part.We detected the 24h urine protein,serum creatinine,blood urea nitrogen and other biochemical changes of these rats in the 7th,14th,21st,28th and 35th day.The renal pathology change was observed by the HE and PAS staining through microscope;the deposition of IgG was observed by immunofluorescence;the podocyte,glomerular basement membrane,and the deposition of immune complex were observed by electron microscope.We found that the 24h urinary protein excretion, serum creatinine,and blood urea nitrogen of glomerulonephritis group was significantly higher（P0.01） than the normal control group at all time points.And renal pathology observation showed there were significantly increase of glomerular cells,the formation of cellular crescent and protein cast,and the necrosis of endothelial cell in glomerulonephritis group.While the diffuse line-like deposition of IgG was observed.We also found an irregular thickening of glomerular basement membrane and foot process fusion while the normal control group showed no change.We successfully established the rat anti-GBM nephritis model and the lesion of this model are similar to the human anti-GBM nephritis lesion,which can be used for further research of anti-GBM nephritis study.