文章摘要
朱宇峰,蔡梦媛,刘昌伟,王庆林,张恩瑞,钱宝梅,张志伟,赵 聃,邱 文,王迎伟.SP1基因在sublytic C5b⁃9促大鼠肾小球系膜细胞增殖中的作用[J].南京医科大学学报,2018,(11):1499~1505
SP1基因在sublytic C5b⁃9促大鼠肾小球系膜细胞增殖中的作用
The role of SP1 gene in sublytic C5b⁃9⁃induced GMC proliferation
投稿时间:2018-05-14  
DOI:10.7655/NYDXBNS20181103
中文关键词: Thy⁃1肾炎(Thy⁃1N)  肾小球系膜细胞(GMCs)  增殖  亚溶解型C5b⁃9(sublytic C5b⁃9)  特异性蛋白1(SP1)
英文关键词: Thy⁃1 nephritis(Thy⁃1N)  glomerular mesangial cells(GMCs)  proliferation  sublytic C5b⁃9  specificity protein 1(SP1)
基金项目:国家自然科学基金(31470853,81471626);江苏省高等学校大学生创新训练计划项目(201710312020Z)
作者单位
朱宇峰 南京医科大学第一临床医学院临床医学系(长学制)江苏 南京 211166 
蔡梦媛 南京医科大学第一临床医学院临床医学系(长学制)江苏 南京 211166 
刘昌伟 南京医科大学第一临床医学院临床医学系(长学制)江苏 南京 211166 
王庆林 南京医科大学第一临床医学院临床医学系(长学制)江苏 南京 211166 
张恩瑞 南京医科大学第一临床医学院临床医学系(长学制)江苏 南京 211166 
钱宝梅 南京医科大学基础医学院免疫学系江苏 南京 211166 
张志伟 南京医科大学基础医学院免疫学系江苏 南京 211166 
赵 聃 南京医科大学基础医学院免疫学系江苏 南京 211166 
邱 文 南京医科大学基础医学院免疫学系江苏 南京 211166 
王迎伟 南京医科大学基础医学院免疫学系江苏 南京 211166 
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中文摘要:
      目的:检查特异性蛋白1(specificity protein 1,SP1)在Thy?1肾炎(Thy?1 nephritis,Thy?1N)大鼠肾组织中和亚溶解型C5b?9(sublytic C5b?9)刺激的大鼠肾小球系膜细胞(glomerular mesangial cells,GMCs)中的表达情况,并探讨SP1表达上调对sublytic C5b?9促大鼠GMCs增殖的作用。方法:复制大鼠Thy?1N模型,同时用sublytic C5b?9刺激体外培养的大鼠GMCs,分别用real?time PCR和Western blot检查Thy?1N大鼠肾组织(体内)和sublytic C5b?9刺激的大鼠GMCs(体外)中SP1基因的mRNA和蛋白表达水平及其表达时相。与此同时,构建SP1基因的表达质粒(pEGFP?N1/SP1?His)和发夹状小干扰RNA质粒(SP1 shRNA,shSP1)。将上述质粒分别转染大鼠GMCs(同时设对照质粒转染组),再给予或不给予sublytic C5b?9刺激。用real?time PCR和Western blot检查SP1表达量,并行CCK?8实验检查GMCs的增殖情况。结果:在Thy?1N大鼠肾组织和sublytic C5b?9刺激的大鼠GMCs中,SP1 mRNA和蛋白表达水平均显著上调,且其体内外表达时相较为一致。体外过表达SP1可显著促进GMCs增殖,而沉默SP1可明显抑制sublytic C5b?9诱导的GMCs增殖反应。结论:Sublytic C5b?9刺激大鼠GMCs后能通过上调SP1表达促进GMCs增殖。
英文摘要:
      Objective:To detect the expression of specificity protein 1(SP1)in the renal tissues of rats with Thy?1 nephritis(Thy?1N)and in the glomerular mesangial cells(GMCs)stimulated with sublytic C5b?9,and investigate the role of SP1 expression in sublytic C5b?9?induced GMC proliferation. Methods:Rat Thy?1N was induced,and the cultured rat GMCs were stimulated with sublytic C5b?9. The expression of SP1 in the renal tissues of rats with Thy?1N(in vivo)and in the GMCs stimulated with sublytic C5b?9(in vivo)was detected by real?time PCR and Western blot,respectively. Meantime,SP1 expression plasmid(pEGFP?N1/SP1?His)and SP1 shRNA(shSP1)were constructed. Above?mentioned plasmids and control plasmids were transfected into rat GMCs followed by sublytic C5b?9 stimulation or not. The expression levels of SP1 were evaluated by real?time PCR and Western blot,and the cellular proliferation was determined by CCK?8. Results:The expression levels of SP1 in the renal tissues of rats with Thy?1N and in the GMCs stimulated with sublytic C5b?9 were markedly increased with similar time points. SP1 overexpression could markedly promote GMCs proliferation,while SP1 knockdown could obviously reduce sublytic C5b?9?induced GMCs proliferation. Conclusion:Sublytic C5b?9 promotes rat GMCs proliferation through up?regulation of SP1.
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