大鼠慢性血清病肾炎模型的改进研究

Improvement Study of a Model of Rat Chronic Serum Sickness Glomerulonephritis

目的 研究提高慢性血清病肾炎模型动物的发病率、降低其制作难度的方法。方法 36只Wistar雌性大鼠分为正常对照组、改进造模组、传统造模组 3组 ,每组 12只。改进组造模方法为 :切除左侧肾脏 ;3mg牛血清白蛋白 (BSA)与佐剂混匀双后足垫注射 ,然后每隔 2周重复皮下多点注射一次 ;足垫注射BSA后 2周开始隔日饮饲含 0 1%BSA的 6mmol L盐酸酸化水 ;BSA免疫注射 3次后测血清抗BSA抗体滴度 ;达到 1∶16后开始每日腹腔注射 3mg的BSA ;3周后 ,腹腔注射 10 0 μg的LPS一次 ;4周后宰杀大鼠。传统造模组按文献方法不做肾切、不饲BSA酸化水、不注射LPS ,其中每日腹腔注射BSA改为每日尾静脉注射 ,其他处理同改进造模组。检测指标有一般情况、肾重指数、肾炎发生率、蛋白尿情况、血生化、病理改变、免疫荧光改变等。结果 与对照组比较 ,两造模组大鼠均出现大量蛋白尿、血白蛋白降低及高脂血症、肾功能降低 ,病理显示系膜细胞增生、炎细胞浸润、大量蛋白管型等 ,免疫荧光显示IgG、C3肾小球内沉积等变化。而改进造模组比传统造模组肾炎发生率高 ,改变更为严重。结论 通过切除大鼠一侧肾脏、饮饲BSA酸化水、腹腔注射LPS、改尾静脉注射BSA为腹腔注射等措施 ,可以降低慢性血清病肾炎模型的制作难度。

Objective To improve a method so as to enhance the disease incidence and reduce the establishing difficulty of the chronic serum sickness glomerulonephritis model in rat.Methods\ 36 rats were divided into three groups with 12 rats each:normal control,improved method and traditional method group.The establishment of improved method group was as follows: the left kidney removed, footpad injection of 3mg BSA emulsified with Freund's adjuvant,then injection subcutaneously every 2 weeks for 3 times.2 weeks after the footpad injection 0.1% 6mmol/L HCl acidified BSA water was given every two days in the drinking water.3 times after BSA injection the anti\|BSA antibody titer in the serum was determined,and if the titer was above 1∶16,then 3mg BSA was given everyday intraperitoneally,3 weeks later 100μg of LPS was given intraperitoneally,and the rats were killed 4 weeks later.The establishment of the traditional group was identical to that in literature.The observed indices were:general situation,kidney index,nephritis rate,proteinuria,blood biochemical indices,pathological changes,and immunofluorescence changes etc.Results\ Compared with normal control group,the two model groups both showed heavy proteinuria,lowered blood albumin,hyperlipemia,lowered kidney function,and the proliferation of mesangial cell,infiltration of inflammatory cells,large amount of protein cast,and the immunofluorescence showed deposition of IgG,IgA and C3 in the mesangial area.And the changes of improved method group were more serious.Conclusion\ Through the described methods we successfully established a model of chronic serum sickness glomerulonephritis with lowered difficulty and enhanced disease incidence.