CAJ | 학술논문

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。
목적：관찰용혈린지산（LPA）대인단핵세포주THP-1세포핵인자-κB p65（NF-κB p65）표체、핵이위급염성세포인자종류배사인자α（TNFα）변화적영향，탐토LPA치동맥죽양경화적궤제。방법이불동농도수평LPA（0~10μM）자격인THP-1세포4 h，혹이LPA 1μM처리THP-1세포불동시간（0~8 h），Westernblot검측THP-1세포핵단백NF-κB p65표체변화，면역형광검측NF-κB p65단백표체핵이위,매련면역법측정세포상청액중세포인자TNF-α수평。장세포여NF-κB억제제가배산분을지（CAPE)）20 mg/L예처리1 h후, LPA 1μM처리4 h후，매련면역법측정세포상청액중TNFα수평。결과 LPA이제량의뢰화시간의뢰적방식유도THP-1세포NF-κB p65표체，촉진NF-κB p65전위도핵내，병촉진TNF-α분비，CAPE억제NF-κB후가현저억제TNF-α분비。결론 LPA가현저촉진THP-1세포N F-κB적표체화활화，촉진염성인자TNFα적석방，N F-κB신호도경부분개도료LPA치죽양경화작용。
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.