溶血磷脂酸对人单核细胞株THP-1细胞核因子-κB表达的影响  

The effect of lysophosphatidie acid on the expression of nuclear factor-kappaB signaling pathway in THP-1cell

作　　者：牛建平[1] 周志斌[2] 宋叶华[1] 林俊[1] 

机构地区：[1]厦门市第二医院神经内科,361021 [2]武汉科技大学附属天佑神经内科,430065

出　　处：《中华脑血管病杂志（电子版）》2013年第5期1-3,7,共4页Chinese Journal of Cerebrovascular Diseases(Electronic Edition)

基　　金：福建省自然科学基金(2010D010)

摘　　要：目的：观察溶血磷脂酸（LPA）对人单核细胞株THP-1细胞核因子-κB p65（NF-κB p65）表达、核移位及炎性细胞因子肿瘤坏死因子α（TNFα）变化的影响，探讨LPA致动脉粥样硬化的机制。方法以不同浓度水平LPA（0~10μM）刺激人THP-1细胞4 h，或以LPA 1μM处理THP-1细胞不同时间（0~8 h），Westernblot检测THP-1细胞核蛋白NF-κB p65表达变化，免疫荧光检测NF-κB p65蛋白表达核移位,酶联免疫法测定细胞上清液中细胞因子TNF-α水平。将细胞予NF-κB抑制剂咖啡酸苯乙酯（CAPE））20 mg/L预处理1 h后, LPA 1μM处理4 h后，酶联免疫法测定细胞上清液中TNFα水平。结果 LPA以剂量依赖和时间依赖的方式诱导THP-1细胞NF-κB p65表达，促进NF-κB p65转位到核内，并促进TNF-α分泌，CAPE抑制NF-κB后可显著抑制TNF-α分泌。结论 LPA可显著促进THP-1细胞N F-κB的表达和活化，促进炎性因子TNFα的释放，N F-κB信号途径部分介导了LPA致粥样硬化作用。Objectives To explore the effect of lysophosphatidie acid（LPA）on the expression and neuclear translocation of NF-κB p65 protein in cell nucleus and the secretion of TNF-α in THP-1cell line, and to further investigate the mechanism of atherosclerosis caused by LPA. Methodes Human THP-1 cells were stimulated with LPA at different concentrations（0~10μM）or at 1 μM for different time durations（0~8 h）, the expression and neuclear translocation of NF-κB p65 in cell nucleus was detected by Westernblot and immunofluorescence method respectively, and the secretion of TNF-αwas detected using a sandwich ELISA. Then THP-1 cell were perincubated with CAPE for 1 h and then stimulated with LPA at 1 μM for 4 h, the expression TNF-α was measured by enzyme-linked immunosorbent assay. Results LPA enhanced the NF-κB p65protein levels in cell nucleus and its transfer to the nucleus. The enhancement effects occurred in concentration- and time-dependent manners. LPA also increased the level of TNF-αin a dose-dependent manner, which can be significantly depressed by CAPE. Conclusions LPA may synchronously increase the expression and activate the NF-κB p65, and promote the secretion of TNF-α. The atherogenic effect of LPA maybe partially mediate through N F-κB pathway.

关 键 词：溶血磷脂酸 THP-1细胞 核因子-ΚB 

分 类 号：R346[医药卫生—基础医学]