【摘要】：目的：探討亞溶解劑量的補(bǔ)體C5b-9復(fù)合物(sublytic C5b-9，SC5b-9)體外能否刺激腎小球系膜細(xì)胞(glomerular mesangial cells，GMCs)上調(diào)增殖細(xì)胞核抗原(proliferation cell nuclear antigen，PCNA)、細(xì)胞外基質(zhì)(extracellular matrix，ECM)成份—纖維粘連蛋白(fibronectin，F(xiàn)N)、α-平滑肌肌動(dòng)蛋白(α-smooth muscle antin，α-SMA)的表達(dá)水平。此外，觀察SC5b-9促進(jìn)GMCs增生的同時(shí)，是否伴隨細(xì)胞核轉(zhuǎn)錄因子-κB(nuclear factor-κB，NF-κB)的活化。方法：(1)體外純化培養(yǎng)GMCs并經(jīng)光鏡、電鏡、細(xì)胞收縮功能實(shí)驗(yàn)(10~(-7)mol／L血管緊張素Ⅱ，ATⅡ)及抗肌動(dòng)蛋白(actin)抗體、抗結(jié)蛋白(desmin)抗體、抗細(xì)胞角蛋白(keratin)抗體染色鑒定。(2)采用確定的SC5b-9刺激培養(yǎng)的GMCs，實(shí)驗(yàn)設(shè)SC5b-9刺激組，另設(shè)抗胸腺細(xì)胞血清(anti-thymocyte serum，ATS)組、滅活人血清組、完全營養(yǎng)液組作為對(duì)照。用RT-PCR技術(shù)檢測刺激40min時(shí)，PCNA、FN mRNA的表達(dá)，免疫組化檢測實(shí)驗(yàn)40min、6h、18hPCNA、α-SMA和FN及細(xì)胞核內(nèi)NF-κB p65表達(dá)情況，同時(shí)應(yīng)用western blot方法測定6h、18h FN蛋白表達(dá)，觀察SC5b-9對(duì)GMCs增生和細(xì)胞外基質(zhì)分泌的影響。(3)體外純培養(yǎng)的GMCs用吡咯啉烷二甲基硫脲(PDTC)處理，實(shí)驗(yàn)分為三組，即SC5b-9刺激組、PDTC+SC5b-9組和完全營養(yǎng)液組，，用RT-PCR分析刺激GMCs 40min后，PCNA、FN mRNA表達(dá)的變化。除此之外，還用免疫組化方法檢查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 偽-smooth muscle actin (偽-smooth muscle antin, 偽-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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