重组人红细胞生成素影响系膜细胞生长与纤维连接蛋白表达的实验研究  

Experimental study of recombinant human erythropoietin up-regulated growth and fibronectin expression in glomerular mesangial cells

作　　者：周书琴[1] 闻萍[1] 高雯[1] 王晓华[1] 方丽[1] 狄佳[1] 江蕾[1] 熊明霞[1] 张飞飞[1] 杨俊伟[1,2] 

机构地区：[1]南京医科大学第二附属医院肾脏病医学中心,江苏南京210003 [2]南京医科大学第一附属医院江苏省糖尿病中心,江苏南京210029

出　　处：《中国药理学通报》2009年第1期60-64,共5页Chinese Pharmacological Bulletin

基　　金：国家重点基础研究发展计划(973计划)重点资助项目;国家自然科学基金资助项目(No30470800,30771010);国家人事部回国人员科研启动基金;江苏省“六大人才”高峰项目;江苏省“科教兴卫”工程医学领军人才资助项目

摘　　要：目的通过观察重组人红细胞生成素(rhEPO)影响肾小球系膜细胞生长,以及表达纤维连接蛋白(FN)的作用,探讨应用rhEPO后对肾组织可能产生的生物学效应。方法培养的大鼠肾小球系膜细胞在不同剂量(1、10、100和1000U.ml-1)rhEPO的作用下,细胞计数和蛋白浓度测定评价细胞是否肥大;蛋白免疫印迹法检测FN蛋白表达的变化。♂CD-1小鼠接受单次(n=8)腹腔注射rhEPO(1000 U.kg-1体重),注射后24 h处死并分离肾小球以检测其FN表达量的变化。结果①rhEPO能够增加系膜细胞蛋白质的含量,以100 U.ml-1工作浓度rhEPO的作用效果最为明显[(1.269±0.382)μg.10-3cellsvs(0.247±0.058)μg.10-3cells;P<0.01)];②rhEPO能够刺激系膜细胞上调FN蛋白的表达,与对照组相比,100 U.ml-1的rhEPO明显增加系膜细胞FN蛋白的表达,随着rhEPO浓度的增加,系膜细胞FN的表达进一步增多,呈剂量依赖性;③间接免疫荧光检测可见rhEPO作用系膜细胞24 h后,可见大量呈网状、条索状,甚至板块状FN蛋白分布于细胞间隙;④单次腹腔注射rhEPO后24 h的肾小球FN增加。结论rhEPO可以造成系膜细胞的肥大,同时诱导系膜细胞增加FN合成,提示rhEPO可以通过影响系膜细胞的功能造成肾小球功能的异常。Aim To investigate the effects of recombi- nant human erythropoietin （rhEPO） on cellular growth and fibronectin （FN） expression in glomerular mesangial cells. Methods rhEPO-induced cellular hypertrophy was quantified by counting cell number and calculating the ratio of cell protein to cell number. Western blot analysis was used to detect the expression of FN induced by rhEPO （1, 10, 100 and 1000 U · ml-1 ）. In vivo studies, male CD-1 mice were administered rhEP0 subcutaneously at a single dose of 1000 U ·kg-1. 24 hours after the injection, the expression FN was assessed by Western blot analysis. Results （1）rhEPO induced hypertrophy of mesangial cells .（2）rhEPO induced FN expression of mesangial cell in a dose-dependent way. Compared to control, 100 U · ml-1 rhEPO significantly enhanced the expression of FN. （3） Indirect immunofluorescence showed that rhE- PO induced large amount deposition of FN in the intercellular space of mesangial cells. （4） In vivo studies, there were marked increase of FN expression in mice received the injection of rhEPO. Conclusions rhEPO induces glomerular mesangial cells hypertrophy and up- regulates the expression of FN. These results suggest rhEPO can induce dysfunction of renal glomerulus through its influence on the function of mesangial cells.

关 键 词：红细胞生成素 系膜细胞 肥大 纤维连接蛋白 

分 类 号：R-332[医药卫生] R322.61