HSF-1抑制HMGB1引起的炎症反应的机制.docVIP

  HSF-1 抑制 HMGB1 引起的炎症反应的 机制# 李杉珊，尹朝奇，罗成群，贺全勇，周建大，卜平元，朱颉，彭晨， 彭浩， 5 10 15 20 25 30 35 40 徐丹** （中南大学湘雅三医院烧伤整形外科，长沙 410013） 摘要：目的 探讨热休克转录因子 1 抑制高迁移率族蛋白 1 引起炎症反应的机制。方法 HSF1 质粒转染 RAW264.7 细胞，1μg/ml HMGB1 刺激空白细胞，转染空质粒细胞，HSF 过表达细 胞 4h 后收集细胞上清。酶联免疫吸附法测定 TNF-α 水平。蛋白免疫印迹法检测 p-p38, p-ERK1/2, p-JNK1/2, p-IKα-B 的表达。非放射性凝胶阻滞检测 NF-κB 与 DNA 结合情况。结 果 （1）过表达 HSF-1 抑制 TNF-α 的表达 4h, 13765,3±402.4 pg/ml vs 9345.2±359.3pg/m l(P<0.01, t=13.9)。（2）过表达 HSF-1 对 MAPK 通路的活化无明显影响（P>0.05, F<1）。 (3) 过表达 HSF-1 对 IKα-B 的磷酸化无明显影响。（4）过表达 HSF-1 抑制 NF-κB 与 DNA 的结 合能力，83.7±9.8 vs 32.3±8.2 (P<0.05, t=5.2) 结论 HSF-1 过表达可以减少 HMGB1 引起的 TNF-α 表达，其分子机制与胞内信号通路的活化无关，但与 NF-κB 和 DNA 的结合能力有关。 关键词：烧伤外科；高迁移率族蛋白 1；热休克转录因子 1；炎症反应 中图分类号：R644 the mechanism of HSF-1 inhibited inflammation induced by HMGB1. Li Shanshan, Yin Chaoqi, Luo Chengqun, He Quanyong, Zhou Jianda, Bu Pingyuan, Zhu Jie, Peng Chen, Peng Hao, Xu Dan (Department of Burns and Plastic, the 3rd Xiangya Hospital, Central South University, ChangSha 410013) Abstract: Objective To discuss the mechanism of heat shock transcription factor 1 inhibited inflammation induced by high mobility group protein B1.Methods Overexpression HSF-1 by transfected plasmid in RAW264.7 cell, then stimulated cell with HMGB1 (1μg/ml), and the supernatant was assayed for TNF-α by ELISA 4h after stimulation. Expressions of p-p38, p-ERK1/2, p-JNK1/2 and p-IKα-B in cell lysates were detected by Western blot. NF-κB binding were detected by electrophoretic mobility shift assay.Results (1) Overexpression HSF-1 inhibited the expression of TNF-α, 13765,3±402.4 pg/ml vs 9345.2±359.3pg/m l(P<0.01, t=13.9).(2) Overexpression HSF-1 did not affect the activity of MAPK（P>0.05, F<1）.(3) Overexpression HSF-1 did not affect the phosphorylation of IKα-B（P>0.05, F<1）.(4) Overexpression HSF-1 inhibited the binding between NF-κB and DNA. 83.7±9.8 vs 32.3±8.2 (P<0.05, t=5.2).Conclusion Overexpression HSF-1 inhibited the expression of TNF-α, the mechanism was associated with the NF-κB binding but not t