肿瘤坏死因子α和白细胞介素1β对内皮细胞妊娠相关血浆蛋白A表达的影响  

Effects of TNF-α and IL-1β on the Expression of Pregnancy-Associated Plasma Protein-A in Endothelial Cells and Its Mechanism

作　　者：边云飞[1] 赵欣[1] 肖传实[1] 

机构地区：[1]山西医科大学第二医院心内科,山西省太原市030001

出　　处：《中国动脉硬化杂志》2009年第4期260-264,共5页Chinese Journal of Arteriosclerosis

基　　金：山西省高校科技研究开发项目(200811070)

摘　　要：目的观察肿瘤坏死因子α和白细胞介素1β对内皮细胞妊娠相关血浆蛋白A表达的影响并探讨其机制。方法原代培养大鼠主动脉内皮细胞,选择生长良好的第3～4代细胞用于实验。实验分组:①空白对照组;②10、20、40、60及100μg/L肿瘤坏死因子α培养细胞24h组;③1、5、10、20及50μg/L白细胞介素1β培养细胞24h组;④60μg/L肿瘤坏死因子α培养细胞2、4、8、16、24及48h组;⑤20μg/L白细胞介素1β培养细胞2、4、8、16、24及48h组;⑥核因子κB抑制剂BAY11-7082干预组:预先用核因子κB抑制剂BAY11-7082(20μmol/L)与内皮细胞共同孵育60min后,再加入60μg/L肿瘤坏死因子α或20μg/L白细胞介素1β作用48h。实验结束后收集细胞及培养上清液,用乳酸脱氢酶试剂盒检测各组培养上清液中乳酸脱氢酶活性,用逆转录聚合酶链反应测定细胞中妊娠相关血浆蛋白AmRNA的表达,用酶联免疫吸附法检测上清液中妊娠相关血浆蛋白A蛋白水平。结果不同浓度肿瘤坏死因子α和白细胞介素1β作用24h后,大鼠内皮细胞妊娠相关血浆蛋白AmRNA及上清液中妊娠相关血浆蛋白A蛋白表达水平随肿瘤坏死因子α和白细胞介素1β浓度的增加而升高(P<0.05);60μg/L肿瘤坏死因子α和20μg/L白细胞介素1β作用于大鼠内皮细胞不同时间,妊娠相关血浆蛋白A表达水平显著高于空白对照组(P<0.05),且随刺激时间的延长,妊娠相关血浆蛋白A的表达逐渐升高;核因子κB抑制剂BAY11-7082作用后,妊娠相关血浆蛋白A表达水平明显降低(P<0.05)。结论肿瘤坏死因子α和白细胞介素1β上调内皮细胞妊娠相关血浆蛋白A的表达,而核转录因子的活化是其对内皮细胞妊娠相关血浆蛋白A表达上调的主要机制。Aim To investigate the effects of tumor necrosis factor-or（TNF-α） and interleukin-1β（IL-1β） on pregnancy-associated plasma protein-A（PAPP-A） expression fin endothelial ceils and analyse its mechanism. Methods The rat aortic endothelial cells were obtained from thoracic aortas and cultured by tissue explant method. Passage 3-4 cells were used in the experiment. Rortic endothelial cells were coincubated with different concentrations of TNF-α（10, 20, 40, 60 and 100μg/L） and IL-1β（1, 5, 10, 20 and 50μg/L） for 24 h or coincubated with 60μg/L TNF-α, 20 μg/L IL-1β for different periods（2, 4, 8, 16, 24 and 48 h）. BAY11-7082（20μmol/L） was pretreated for 60 min before rortic endothelial cells were coincubated with 60μg/L TNF-α, 20μg/L IL-1β for 48 h. LDH activity in superna-tant was detected. PAPP-A mRNA and protein expression were detected by RT-PCR or ELISA. Results After the intervention of TNF-α or IL-1β for 24 h, the levels of PAPP-A expression increased with the concentration raise （P〈0.05） ; Cytokines stimulated PAPP-A expression in the two groups （P〈0.05） ; BAY11-7082 inhibited PAPP-A expression stimulated by TNF-α and IL-1β. Conclusions TNF-α and IL-1β may stimulate the expression of PAPP-A, nuclear factor-κB（NF-κB） activation may be the primary mediator of cytokines stimulated PAPP-A expression.

关 键 词：妊娠相关血浆蛋白A 肿瘤坏死因子Α 白细胞介素1Β 内皮细胞 核因子ΚB 

分 类 号：R5[医药卫生—内科学]