【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。更多还原

【Abstract】 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.更多还原