亚溶解剂量的补体C5b-9复合物诱导肾小球系膜细胞增生及其NF-κB活化的作用  

Effects of sublytic complement C5b-9 complex on the proliferation of glomerular mesangial cell associated with NF-κB activation

作　　者：佟建霞[1] 王迎伟[1] 邱文[1] 高玲娟[1] 徐娟[1] 吴宜琴[1] 徐静华[1] 徐闻欢[1] 

机构地区：[1]南京医科大学微生物与免疫学系,江苏南京210029

出　　处：《南京医科大学学报（自然科学版）》2006年第4期230-233,241,F0002,共6页Journal of Nanjing Medical University(Natural Sciences)

基　　金：国家自然科学基金课题(30471615);江苏省教育厅自然科学基金重点课题(03KJA310074);南京医科大学创新基金资助课题(CX2002006)

摘　　要：目的:研究亚溶解剂量补体C5b-9(SC5b-9)复合物刺激大鼠肾小球系膜细胞(GMCs)增生和细胞外基质(ECM)分泌的作用,并探讨NF-κB核转录因子是否介入SC5b-9的促增生效应。方法:体外纯培养大鼠GMCs,并行光镜、电镜及功能鉴定。质控SC5b-9后,将GMCs分为5组,即SC5b-9刺激组、SC5b-9+PDTC组(吡咯啉烷二甲基硫脲)、ATS组、灭活人血清组及完全营养液组。应用RT-PCR检测40minPCNA和FNmRNA水平;同时应用免疫组化检测18hGMCs增殖细胞核抗原(PCNA)、平滑肌肌动蛋白(α-SMA)和纤维粘连蛋白(FN)及细胞核内NF-κBp65表达情况。结果:实验40min,SC5b-9刺激组PCNA和FNmRNA表达上调,PDTC处理后表达则明显下降,其余3组PCNA均呈阴性,另3组FN则均有一定基础表达。SC5b-9刺激组,GMCs免疫组化显示:PCNA、α-SMA、FN、NF-κBp65表达增加,PDTC处理后则4者表达明显下调,其余各组,PCNA、α-SMA、NF-κBp65均呈阴性,FN亦有一定的基础表达。结论:SC5b-9可能通过激活NF-κB促进GMCs增生和ECM分泌。Objective： To explore the effects of sublytic C5b-9 （SC5b-9）complex on the proliferation of rat glomerular mesangial cells （GMCs） and the secretion of extracellular matrix （ECM） and whether NF-κB is involved in or not. Methods： The rat GMCs lines were cultured in vitro and identified through light microscope, electron microscope and functional test. After the formation of sublytic C5b-9, the GMCs were then divided into SC5b-9 group, SC5b-9 ＋ PDTC group, ATS group, inactivation human serum group and cultured medium group. The mRNA of both PCNA and FN was examined by RT-PCR at 40 minutes. Meanwhile the cellular expression of PCNA, α-SMA and FN as well as NF-κB p65 was examined by immunohistochemical staining at 18 hours. Results： The RT-PCR results of PCNA and FN mRNA were up-regulated whereas PDTC significantly reduced these increases induced by SC5b-9 at 40 minutes. In other groups PCNA were negative, FN had basic expression. The addition of SC5b-9 to GMCs apparently increased the expression of PCNA, α-SMA, FN and NF-κB p65 whereas PDTC significantly reduced these increases induced by SC5b-9. Immunohistochemical staining for PCNA,α-SMA and NF-κB p65 were negative in other groups （FN had basic expression）. Conclusion： SC5b-9 may promote the proliferation of GMCs and the secretion of ECM through activating NF-κB.

关 键 词：亚溶解剂量补体C5b-9 肾小球系膜细胞 增殖细胞核抗原 平滑肌肌动蛋白 纤维粘连蛋白 核因子-κB 

分 类 号：R692[医药卫生—泌尿科学]