摘要
目的探讨人THP-1单核细胞中白细胞介素6(IL-6)/信号转导与转录激活子3(STAT3)通路与环氧化酶2(COX-2)表达之间的关系。方法以人THP-1单核细胞株作为研究对象,分别设置0至48 h不同时间点,检测IL-6对STAT3磷酸化和COX-2表达的时效性影响。用100μmol/L S3I-201(STAT3通路选择性抑制剂)对THP-1单核细胞预处理24 h,再经10μg/L IL-6作用相应时间,将THP-1单核细胞分为4组:对照组、S3I-201组、IL-6组及IL-6+S3I-201组,检测STAT3磷酸化及COX-2表达水平变化。采用实时荧光定量PCR方法检测THP-1单核细胞COX-2的mRNA表达量,Western blot方法检测THP-1单核细胞STAT3磷酸化水平与COX-2的蛋白表达量。结果THP-1单核细胞经IL-6作用后STAT3的磷酸化及COX-2的表达均出现时效性激活,IL-6刺激5 min后STAT3磷酸化水平即显著增加(P<0.001),同时COX-2 mRNA和蛋白水平均明显上调,分别于1 h和2 h达到峰值(P<0.001)。与对照组相比,S3I-201组COX-2 mRNA及蛋白表达水平均降低(P<0.01);与IL-6组相比,IL-6+S3I-201组STAT3磷酸化水平下降(P<0.001),COX-2 mRNA表达水平降低(P<0.001),同时COX-2蛋白表达受到明显抑制(P<0.05)。结论 IL-6能够激活THP-1单核细胞STAT3通路,其可能通过催化下游COX-2的表达影响动脉粥样硬化性疾病进程。
Aim To investigate the relationship between the interleukin-6（ IL-6） /signal transducer and activator of transcription 3（ STAT3） signaling pathway and cyclooxygenase-2（ COX-2） expression in THP-1 monocytes. Methods Human THP-1 monocyte was used as the research cell,and the time-dependent expressions of STAT3 phosphorylation and COX-2 were detected after IL-6 treatment for 0 to 48 hours. THP-1 monocytes were pretreated with 100 μmol / L S3I-201（ an specific inhibitor of STAT3 signaling） for 24 hours and then treated with 10 μg / L IL-6 for certain time. THP-1monocytes were divided into 4 groups： control group,S3I-201 group,IL-6 group and IL-6 ＋ S3I-201 group,then the changes of STAT3 phosphorylation and COX-2 expression were detected. COX-2 mRNA expression was detected by real-time fluorescence quantitative PCR. The levels of STAT3 phosphorylation and COX-2 protein expression were determinated by Western blot. Results IL-6 could obviously induce STAT3 phosphorylation and COX-2 expression via a time-dependent manner in THP-1 monocytes. Phosphorylation level of STAT3 increased after IL-6 stimulation for only 5 minutes（ P〈0. 001）,meanwhile,COX-2 mRNA and protein expressions was significantly upregulated,reaching peak at 1 h and 2 h respectively（ P〈0. 001）. Compared with control group,COX-2 mRNA and protein expressions were both markedly suppressed in S3I-201 group（ P〈0. 01）. Compared with IL-6 group,phosphorylated STAT3 level was downregulated in IL-6 ＋ S3I-201 group（ P〈0. 001）,and COX-2 mRNA expression was also decreased（ P〈0. 001）,with COX-2 protein expression clearly suppressed（ P〈0. 05）. Conclusion IL-6 is capable of activating the STAT3 pathway in THP-1monocytes,which may play a role in COX-2 expression and further affect the process of atherosclerotic disease.
出处
《中国动脉硬化杂志》
CAS
北大核心
2016年第3期251-255,260,共6页
Chinese Journal of Arteriosclerosis
基金
上海市科学技术委员会基金(124119a6800)
