抗α1-肾上腺素受体抗体介导糖尿病大鼠肾损伤的模型建立

Model of Renal Microvascular Disease in Diabetes Mellitus Rats Mediated by α1-Adrenergic Receptor Antibodies

目的建立具有激动样活性的抗α1肾上腺素受体抗体(α1-adrenergic receptor autoantibody,α1-R Ab)大鼠模型,观察该抗体对大鼠肾脏功能24 h尿蛋白(24 h urinary protein,24 h Upro)和血肌酐(serum creatinin,SCr)、肾脏结构及转化生长因子β1(transforming growth factorβ1,TGF-β1)表达的影响。方法 Wistar大鼠64只,随机分为4组:①健康鼠空白对照组;②健康鼠α1-R Ab介导组;③糖尿病(diabetes mellitus,DM)鼠无α1-R Ab介导组;④DM鼠α1-RAb介导组。采用腹腔注射链脲佐霉素(streptozotocin,STZ)法复制糖尿病大鼠模型,造模成功后持续观察16周;用α1肾上腺素受体胞外第二肽段合成肽免疫②、④组Wistar大鼠(注射α1-R Ab,100μg/100 g体质量,分别在0、4、8、12、16周鼠尾静脉一次性注射);应用酶联免疫吸附测定(enzyme-linked immunosorbent assay,ELISA)技术检测α1-R Ab;放射免疫法测定24 h Upro水平,苦味酸法测定SCr水平;免疫组织化学染色法(immunohistochemistry,IHC)检测肾脏组织TGF-β1表达;用光镜和电镜观察肾脏病理变化。结果①DM鼠α1-R Ab介导组24 h Upro和SCr水平均明显高于DM鼠无α1-R Ab介导组(均P<0.01)。健康鼠α1-R Ab介导组24h Upro明显升高,与健康鼠空白对照组比较,差异有统计学意义(P<0.05)。②电镜下DM鼠α1-R Ab介导组肾组织微观结构严重破坏,健康鼠α1-R Ab介导组及DM鼠无α1-RAb组可见轻度异常,健康鼠空白对照组未见明显异常。③α1-R Ab介导组DM鼠组肾皮质中TGF-β1的表达较无α1-RAb介导组明显增多,有统计学意义(P<0.01)。结论α1-R Ab介导的鼠体肾皮质中TGF-β1表达增加,糖尿病大鼠肾脏微血管损伤与α1-R Ab介导的免疫反应有关。

Objective To establish diabetes mellitus （DM） rat model with α1-adrenergic receptor autoantibodies （α1-R Ab）, and investigate the effects ofα1-R Ab on the 24 hour urinary protein （24hUpro）, serum creatinine （SCr）, kidney structures and the expression of transforming growth factor α1 （TGF-β1）. Methods A total of 64 Wistar rats di- vided into blank control group with healthy rats, α1-R Ab-mediated healthy rat group, non-mediated DM rat group, and α1 -R Ab -mediated DM rat group. Peritoneal injection of streptozotocin was utilized to induce DM model rats. Observation persisted for 16 weeks after successful model establishment. Theα1-R Ab-mediated healthy rat group and α1-R Ab-media- ted DM rat group were injected with α1-RAb by tail vein at 0, 4, 8, 12, 16 weeks, 100 tlg/100 g by weight.α1-RAb was detected by enzyme-linked immunosorbent assay （ELISA）. 24 h Upro was detected by radioimmunoassay, and Scr was tedted by picric acid method. TGF-β1 expression was tested by immunohistochemistry （IHC）. The changes of kidney structures with light scope and electronic microscope. Results （1）The levels of 24 h Upro and SCr in α1-R Ab-mediated DM rat group were higher than those in the non-mediated DM rat group （both P〈0. 01）. Compared with the blank con- trol group, 24 h Upro of the healthy α1-R Ab-mediated rat group was significantly increased （P〈0. 05）. （2）Under the e- lectronic microscope, serious destruction was seen in α1 -R Ab-mediated DM rat group. Slight abnormality was seen in α1- R Ab-mediated healthy rat group and non-mediated DM rat group. No abnormality was seen in the blank control group. （3） The expression of TGF-β1 in renal cortex of the α1-R Ab-mediated DM rat group was significantly higher than that in the non-mediated group （P〈0. 01）. Conclusion The expression of TGF-β1 in DM rats mediated by α1-R Ab is increased. The injury of renal capillary of the DM rats is related with the immunity reaction mediated by α1-R Ab.