低剂量FasL诱导bEnd.3细胞增殖及机制研究  

Proliferation and Mechanism of bEnd.3 Induced by Low Dose FasL

作　　者：张春兵[1] 张明顺[2] 滕凤猛[1] 高峰[1] 

机构地区：[1]南京中医药大学附属医院,江苏南京210029 [2]南京医科大学微生物和免疫教研室,江苏南京210029

出　　处：《南京中医药大学学报》2013年第4期355-358,共4页Journal of Nanjing University of Traditional Chinese Medicine

基　　金：江苏省自然科学基金(BE2010768);国家自然科学基金(81171659)

摘　　要：目的探讨低剂量FasL诱导小鼠脑微血管内皮细胞bEnd.3增殖及其可能机制。方法以500～0.015 6ng/mL 1/2倍比稀释共9个浓度梯度FasL干预bEnd.3细胞培养48h,CCK-8检测bEnd.3细胞增殖能力,ELISA检测bEnd.3细胞分泌表达VEGF能力,RNAi抑制Fas表达,Western blotting检测FADD、FLIP、TRAF蛋白表达水平,EMSA检测NF?κB蛋白表达水平。结果 0.156ng/mL FasL干预bEnd.3细胞,可诱导细胞增殖明显增加(P<0.05),bEnd.3细胞分泌表达VEGF能力明显增加(P<0.05),FADD、FLIP、TRAF、NF?κB蛋白表达水平明显增加(P<0.05);RNAi抑制Fas基因后,细胞增殖无明显变化(P>0.05),FADD、FLIP、TRAF蛋白表达水平明显下降(P<0.05),NF?κB蛋白表达水平无明显变化(P>0.05)。结论低剂量FasL可诱导bEnd.3细胞增殖,FADD-FLIP-TRAF-NF?κB信号传导途径是其机制之一。OBJECTIVE To investigate the proliferation and possible mechanism of rat＇s cerebral microvascular endothelial cells induced by low dose FasL.METHODS 9 FasL of different concentration gradients was diluted 500～0.015 6 ng/mL 1/2multiple proportion and the cell culture of bEnd.3 was interfered by 48 h.The cell proliferation ability of bEnd.3 was tested by CCK-8.The VEGF cell secreted expression ability of bEnd.3 was tested by ELISA.Fas Expression was inhibited by RNAi.The protein express levels of FADD,FLIP and TRAF were tested by Westernblot,and the protein express level of NF-κB was tested by EMSA.RESULTS bEnd.3 cell interfered by 0.156 ng/mLFasL could obviously induce the cell proliferation to increase（P＜0.05）.VEGF cell secreted expression ability of bEnd.3 was markedly increased（P＜0.05）.The protein express levels of FADD,FLIP,TRAF and NF-κB increased significantly（P＜0.05） ; The cell proliferation and protein express levels of NF-κB did not change significantly although Fas gene was inhibited by RNAi（P＞0.05）.The protein express levels of FADD,FLIP,TRAF decreased obviously when Fas gene was inhibited by RNAi（P＜0.05）.CONCLUSION Low dose FasL can induce the cell proliferation of bEnd.3,and FADD-FLIP-TRAF-NF-κB signal transduction pathway is one of its mechanisms.

关 键 词：细胞增殖 FAS-FASL RNAI 

分 类 号：R285.5[医药卫生—中药学]