高迁移率族蛋白B1诱导巨噬细胞Janus激酶/信号转导及转录激活子通路活化的研究

High mobility group box - 1 protein activates Janus kinase - signal transducer and activator of transcription pathway in rat peritoneal macrophages

目的 初步探讨高迁移率族蛋白B1(HMGB1)致炎效应的信号转导机制。方法 清洁级雄性Wistar大鼠,取其腹腔巨噬细胞,培养3 d后以10 mg/L HMGB1刺激。刺激完毕后直接在培养瓶中裂解细胞,分别采用免疫沉淀、免疫印迹法和凝胶阻滞分析等技术观察不同时间点Janus激酶2(JAK2)、信号转导及转录激活子-1(STAT1)以及STAT3的活化情况。结果 HMGB1可诱导大鼠腹腔巨噬细胞STAT1、STAT3在短时间内(2 h)活化,其中STAT3活化最为迅速,10 min即可达到活化高峰。但:HMGB1不能在短时间内(2 h)诱导JAK2活化。结论 JAK/STAT途径可能参与了HMGB1致炎效应的信号转导机制。

Objective To investigate the potential signal transduction mechanism in high mobility group box - 1 protein (HMGB1)-induced inflammatory response in rat peritoneal macrophages. Methods Peritoneal macrophages obtained from male Wistar rats were incubated for 3 days before they were stimulated by HMGB1 (10μg/ml). At various time points after HMGB1 stimulation, macrophages were denatured directly in cell culture flasks to detect activation of Janus kinase - 2 (JAK2), signal transducer and activator of transcription - 1 (STAT1) and STATS by immunoprecipitation, Western blotting and electrophoretic mobility shift assay, respectively. Results HMGB1 stimulation could activate STAT1 and STAT3 in peritoneal macrophages in 2 hours, among them the activation of STAT3 appeared to be the quickest, peaking as early as 10 minutes after stimulation. But no marked change in JAK2 activity was observed within 2 hours following HMGB1 stimulation. Conclusion These data suggest that Janus kinase - signal transducer and activator of transcription pathway might be involved in regulation of HMGB1 - induced inflammatory response in peritoneal macrophages.