IgA肾病小鼠模型的建立与鉴定

IgA肾病小鼠模型的建立与鉴定【摘要】目的建立小鼠IgA肾病模型，为临床研究IgA肾病提供实验依据。方法昆明种小鼠30只，分为正常对照组(10只)、模型组(20只)。采用口服牛血清白蛋白(BSA)联合尾静脉注射葡萄球菌肠毒素B(SEB)的方法制作小鼠IgA肾病模型。第12周末收集新鲜尿液，显微镜下尿红细胞定性检验;摘眼球取血，检测血肌酐(Scr)、尿素氮(BUN);光镜法观察肾小球系膜细胞及基质变化;电镜法检测系膜区有无电子致密物沉积;免疫荧光法观察系膜区IgA沉积情况。结果第12周末，模型组小鼠均出现不同程度的血尿;Scr、BUN较正常对照组显著增高(P<0.01);光镜下系膜细胞及基质明显增生;电镜下系膜区电子致密物沉积;免疫荧光示系膜区大量的IgA沉积，而正常对照组则无上述表现。结论 IgA肾病小鼠模型效果良好，病理、电镜及临床指标与人类IgA肾病临床病理相似。

【关键词】 IgA肾病;模型;血尿;免疫荧光;小鼠

Abstract: Objective To establish mouse model of IgA nephropathy in order to provide experiment basis for the clinical research of IgA nephropathy. Methods 30 Kunming mice were randomly assigned into the control group (n=10) and the model group (n=20). Mouse model of IgA nephropathy was established by the method of oral intake of bovine serum albumin

(BSA) together with injection of staphylococcus enterotoxin B (SEB) via the caudal vein. At the end of the 12 weeks, fresh urine was gathered for the quantitative test of hematuria; blood was taken through orbital sinus after eyeball extirpation to determine the serum creatinine (Scr) and blood urea nitrogen (BUN); the changes of mesangial cells and matrix in the kidneys were observed by light microscopy; electron microscopy was employed to detect electron-dense deposits in the mesangial area; the expression of IgA deposition was measured by immunofluorescence staining in the mesangial area. Results By the end of 12 weeks, there were different degrees of hematuria in model group mice. BUN and Scr levels in model group were significantly higher than in the normal control group (P<0.01). Light microscopy showed evident proliferation of mesangial cells and matrix. Electron microscopy demonstrated a high electron dense deposition in the mesangial area, while the above abnormalities were not observed in the normal control group.Conclusion The mouse model of IgA nephropathy proves effective, with pathological electron microscopic and clinical parameters similar to the clinical pathology of human IgA nephropathy.

Key words: IgA nephropathy; model; hematuria; immunofluoresence staining; mice

IgA肾病世界范围内广发的一种系膜增生型肾小球肾炎，占原发性肾小球疾病的10%～20%，中国、日本、新加坡等发病率较高，其中有20%～30%的患者进展到肾衰[1]。建立与人类临床病理生理相似的IgA肾病动物模型，对研究IgA肾病的发病机理、临床治疗等具有重要的作用。本实验采用口服牛血清白蛋白联合尾静脉注射葡萄球菌肠毒素的方法制作IgA肾病小鼠模型，为进一步探索IgA肾病提供实验依据。

1 材料和方法

1.1 实验材料

1.1.1 实验动物昆明种小鼠30只，鼠龄(10-12)周，体重20～26 g，雄性，动物编号SCXK(苏)2005-0005，由徐州医学院实验动物中心提供。

1.1.2 主要实验试剂和设备牛血清白蛋白(BSA,上海明睿生物有限公司，产品批号RF101-010);葡萄球菌肠毒素B(SEB,北京博迈

世纪生物技术有限公司);兔抗小鼠IgA抗体(北京博奥森生物技术有限公司，抗体货号bs-0774R); 异硫氰酸荧光素(FITC)-标记的羊抗兔IgG(Millipore corporation); MicromHM340石蜡切片机;光学显微镜(日本Carton);日本Sysmex CA-1500自动生化分析仪;电镜(H-600)，切片机(LKBV型)。