文章摘要
赵 聃,李 妍,周建博,张 婧,邱 文,王迎伟.大鼠Thy-1肾炎增殖病变及sublytic C5b-9致其肾小球系膜细胞增生的实验研究[J].南京医科大学学报,2012,(10):1343~1350
大鼠Thy-1肾炎增殖病变及sublytic C5b-9致其肾小球系膜细胞增生的实验研究
The proliferative lesions of rat Thy-1 nephritis and the role of sublytic C5b-9 in glomerular mesangial cell proliferation
投稿时间:2012-04-29  
DOI:10.7655
中文关键词: Thy-1肾炎  肾小球系膜细胞  增殖  亚溶解型C5b-9复合物
英文关键词: Thy-1 nephritis  glomerular mesangial cell  proliferation  sublytic C5b-9 complexes
基金项目:国家自然科学基金资助项目(31000396, 81072402);江苏省高校自然科学研究(10KJB310006);南京医科大学科技发展基金面上项目(09NJMUM003);南京医科大学基础医学院青年教师培养基金(09JC007)
作者单位
赵 聃 南京医科大学微生物与免疫学系,江苏 南京 210029 
李 妍 南京医科大学微生物与免疫学系,江苏 南京 210029 
周建博 南京医科大学微生物与免疫学系,江苏 南京 210029 
张 婧 南京医科大学微生物与免疫学系,江苏 南京 210029 
邱 文 南京医科大学微生物与免疫学系,江苏 南京 210029 
王迎伟 南京医科大学微生物与免疫学系,江苏 南京 210029 
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中文摘要:
      目的:检查大鼠Thy-1肾炎(Thy-1 nephritis,Thy-1N)肾小球系膜细胞(glomerular mesangial cell,GMC)的增生病变及亚溶解型(sublytic)C5b-9复合物在GMC增殖中的作用?方法:复制大鼠Thy-1N模型,分别于注射Thy-1抗体后18 h?3 d和7 d时,取肾组织,用real-time PCR和Western blot方法检查其肾组织中细胞增殖指标,即细胞周期蛋白D2(cyclin D2)和增殖细胞核抗原(proliferative cell nuclear antigen,PCNA)的mRNA丰度及蛋白表达水平?用免疫荧光观察肾小球中C5b-9复合物的沉积?同时,使用BrdU掺入法?光镜及电镜观察大鼠肾小球细胞的增殖情况?此外,体外培养大鼠GMC,进行不同分组处理后再给予sublytic C5b-9刺激,用3H-胸腺嘧啶核苷酸掺入法(3H-thymidine incorporation,3H-TdR)检查GMC的增殖情况?结果:大鼠Thy-1N病变18 h,肾组织中cyclin D2?PCNA表达和肾内BrdU阳性细胞数均开始升高,实验3 d和7 d,其表达进一步提高?同时肾小球内可见C5b-9复合物沉积,部分C5b-9包绕的肾细胞形态完好?另形态学观察还发现,实验18 h,部分GMC溶解坏死,而实验3 d,GMC数量明显增多,7 d增加更为显著?体外实验中,大鼠GMC在sublytic C5b-9刺激后18 h,其细胞增殖水平显著增高,而用sublytic C5b-9刺激GMC产生的培养上清处理正常的GMC后,亦能轻度刺激GMC增殖?结论:大鼠Thy-1N肾小球病变存在早期增殖和继发增生的病理改变,而GMC的早期增殖可能与sublytic C5b-9作用有关?
英文摘要:
      Objective:To investigate the proliferative lesions of glomerular mesangial cell (GMC) in Thy-1 nephritis (Thy-1N) rats and the role of sublytic C5b-9 complexes in GMC proliferation. Methods:Thy-1N rat model was constructed. The expression of cyclin D2,proliferative cell nuclear antigen(PCNA) in the renal tissues of rats at mRNA and protein levels were observed at 18 h,on day 3 (3 d) and day 7 (7 d) after Thy-1 Ab injection. The deposition of C5b-9 was detected in the glomerulus. Meanwhile,the proliferation of glomerular cells in rats was assessed by BrdU incorporation,light microscopy(LM) and electron microscopy (EM). Furthermore,rat GMC were cultured and divided into different groups,and then the proliferation of GMC exposed to sublytic C5b-9 was evaluated by 3H-thymidine incorporation(3H-TdR) analysis. Results:The expression of cyclin D2 and PCNA in the renal tissues and the number of glomerular BrdU-positive cells became increasing at 18 h in Thy-1N rats,and continue increasing on 3 d and 7 d. The deposition of C5b-9 was found in the glomerulus,and some cells were intact. Moreover,the number of GMC and the secretion of ECM were increased on 3d and became obvious on 7 d under LM and EM. The proliferation of GMC was increased at 18 h after sublytic C5b-9 stimulation in vitro. The supernatant of GMC induced by sublytic C5b-9 could also promote GMC proliferation of rat. Conclusion:Glomerular lesions in rats with Thy-1N include early and secondary GMC proliferation,while sublytic C5b-9 complexes might be involved in early GMC proliferation.
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