亚溶解剂量补体C5b-9复合物诱导GMCs增生和ECM分泌及其NF-κB活化的作用

发布时间：2018-11-17 13:48

【摘要】：目的：探讨亚溶解剂量的补体C5b-9复合物(sublytic C5b-9，SC5b-9)体外能否刺激肾小球系膜细胞(glomerular mesangial cells，GMCs)上调增殖细胞核抗原(proliferation cell nuclear antigen，PCNA)、细胞外基质(extracellular matrix，ECM)成份—纤维粘连蛋白(fibronectin，FN)、α-平滑肌肌动蛋白(α-smooth muscle antin，α-SMA)的表达水平。此外，观察SC5b-9促进GMCs增生的同时，是否伴随细胞核转录因子-κB(nuclear factor-κB，NF-κB)的活化。方法：(1)体外纯化培养GMCs并经光镜、电镜、细胞收缩功能实验(10~(-7)mol／L血管紧张素Ⅱ，ATⅡ)及抗肌动蛋白(actin)抗体、抗结蛋白(desmin)抗体、抗细胞角蛋白(keratin)抗体染色鉴定。(2)采用确定的SC5b-9刺激培养的GMCs，实验设SC5b-9刺激组，另设抗胸腺细胞血清(anti-thymocyte serum，ATS)组、灭活人血清组、完全营养液组作为对照。用RT-PCR技术检测刺激40min时，PCNA、FN mRNA的表达，免疫组化检测实验40min、6h、18hPCNA、α-SMA和FN及细胞核内NF-κB p65表达情况，同时应用western blot方法测定6h、18h FN蛋白表达，观察SC5b-9对GMCs增生和细胞外基质分泌的影响。(3)体外纯培养的GMCs用吡咯啉烷二甲基硫脲(PDTC)处理，实验分为三组，即SC5b-9刺激组、PDTC+SC5b-9组和完全营养液组，，用RT-PCR分析刺激GMCs 40min后，PCNA、FN mRNA表达的变化。除此之外，还用免疫组化方法检查18h PCNA、α-SMA和FN
[Abstract]:Objective: to investigate whether sublytic dose of complement C5b-9 complex (sublytic C5b-9 SC5b-9) can stimulate glomerular Mesangial cell (glomerular mesangial cells,GMCs to up-regulate proliferating cell nuclear antigen (proliferation cell nuclear antigen,PCNA) and extracellular matrix (extracellular matrix, ECM-fibronectin (fibronectin,FN) and 伪-smooth muscle actin (伪-smooth muscle antin, 伪-SMA). In addition, whether SC5b-9 promotes the proliferation of GMCs is accompanied by the activation of nuclear transcription factor-魏-B (nuclear factor- 魏 B-魏 B (NF- 魏 B). Methods: (1) GMCs was purified and cultured in vitro, and then purified by light microscope, electron microscope, cell contraction function test (10 ~ (-7) mol/L angiotensin 鈪

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