MAPK信号通路调节卷烟烟气诱导的炎症因子的释放

摘要

为评估卷烟烟气对人支气管上皮细胞(BEAS-2B)的毒性效应,使用细胞计数试剂盒(CCK-8)对BEAS-2B细胞进行检测,使用光学显微镜对细胞形态进行观察,采用Western Blot实验检测丝裂原活化蛋白激酶(MAPK)家族的细胞外信号调节激酶1/2(ERK1/2)、c-Jun氨基末端激酶1/2(JNK1/2)和p38的磷酸化水平的变化,采用酶联免疫吸附实验检测炎症因子白介素-1β(IL-1β)、IL-6和IL-8的释放水平.结果表明:①卷烟烟气染毒会显著降低BEAS-2B细胞的存活率.②卷烟烟气染毒会激活磷酸化的ERK1/2、JNK1/2和p38信号通路,并且会诱导炎症因子IL-1β、IL-6和IL-8释放水平升高.③特异性抑制剂分别抑制ERK1/2、JNK1/2和p38信号通路后,炎症因子释放水平显著下降.卷烟烟气诱导的炎症受MAPK信号通路的调节,该结果可为卷烟烟气诱导的肺部炎症相关疾病的机制研究提供参考.%To evaluate the toxic effects of cigarette smoke on human bronchial epithelial cells (BEAS-2B cells), the viability of BEAS-2B cells was detected with a cell counting kit (CCK-8) and the cell morphology was observed under an optical microscopy. Western Blot assay was conducted to detect the expression variations of extracellular-regulated protein kinases 1/2 (ERK1/2) and c-Jun N-terminal kinase (JNK1/2) and the phosphorylation level of p38. The release levels of inflammatory cytokines interleukin-(IL-)1 β, IL-6 and IL-8 were detected by enzyme-linked immunosorbent assay. The results showed that: 1) The viability of BEAS-2B cells significantly decreased after exposure to cigarette smoke. 2) Exposure to cigarette smoke activated the expression of phospho-(P-)ERK1/2, P-JNK1/2, P-p38 pathways and induced the release of inflammatory cytokines IL-1 β, IL-6 and IL-8. 3) The release levels of inflammatory cytokines decreased significantly after ERK1/2, JNK1/2 and p38 pathways’being inhibited by specific inhibitors. In conclusion, the inflammation induced by cigarette smoke is regulated by mitogen-activated protein kinase (MAPK) pathways. These results provide a reference for the mechanism analysis of pulmonary inflammatory diseases induced by cigarette smoke.