HSF-1抑制HMGB1所致炎症反应的作用及机制  

HSF-1 inhibiting inflammatory response induced by HMGB1 and its mechanism

作　　者：尹朝奇[1] 罗成群[1] 周建大[1] 贺全勇[1] 朱颉[1] 李萍[1] 陈铁夫[1] 彭浩[1] 徐阳成[1] 陈佳[1] 

机构地区：[1]中南大学湘雅三医院烧伤整形科,湖南长沙410013

出　　处：《中国普通外科杂志》2014年第9期1207-1212,共6页China Journal of General Surgery

基　　金：高等学校博士学科点专项科研基金资助项目(20090162110020);湖南省科技厅科技计划资助项目(2011sk3243)

摘　　要：目的:探讨热休克转录因子1(HSF-1)抑制高迁移率族蛋白1(HMGB1)引起炎症反应的作用及机制。方法:RAW264.7细胞分别转染含HSF-1的质粒(HSF-1过表达组)和空白质粒(阴性对照组)后,以无处理的RAW264.7细胞作为空白对照组,用Western blot法检测各组细胞HSF-1蛋白的表达;将以上3组细胞分别用HMGB1(1μg/mL)刺激后,用ELISA法测定TNF-α水平、Western blot法检测丝裂原活化蛋白激酶家族(MAPK)通路和NF-κB通路相关蛋白的表达、非放射性凝胶阻滞(EMSA)检测NF-κB与DNA结合活性。结果:Western blot结果显示,HSF-1过表达组HSF-1蛋白表达量较阴性对照组与空白对照组明显升高(均P<0.05),而后两组间HSF-1蛋白表达量无统计学差异(P>0.05);HMGB1作用4 h,HSF-1过表达组TNF-α水平较阴性对照组与空白对照组明显降低(均P<0.05),而后两组间无统计学差异(P>0.05);HMGB1作用不同时间后,各组细胞MAPK通路相关蛋白p-ERK、p-JNK、p-p38以及NF-κB通路相关蛋白p-IKα-B的表达均无统计学差异(均P>0.05);EMSA结果显示,HMGB1作用1 h后,HSF-1过表达组NF-κB灰度值明显低于阴性对照组与空白对照组(均P<0.05),而后两组间无统计学差异(P>0.05)。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MAPK通路的活化无关,但与NF-κB和DNA的结合能力有关。Objective： To investigate the inhibitory effect of heat shock transcription factor I （HSF-1） on the inflammatory response induced by high-mobility group box- 1 protein （HMGB 1） and the mechanism. Methods： RAW264.7 cells were transfected with plasmid bearing HSF-1 gene （HSF-1 overexpression group）or empty plasmid （negative control group） respectively, using the untreated RAW264.7 cells as blank control group, and then the HSF-1 protein expression in each group of cells was determined byWestern blot analysis. The above three groups of cells were stimulated with HMGB1 （1 μg/mL）, and then in each groups of cells, the TNF-α levels were measured by ELISA assa）5 the proteins associated with mitogen-activated p and NF-kB pathways were detected by Western blot analysis, and NF-kB DNA binding rotein kinases （MAPK） activity was analyzed by electrophoretic mobility shift assay （EMSA）, respectively. Results： Western blot analysis showed that the expression level of HSF-1 protein in HSF-1 overexpression group was significantly increased compared with negative control group or blank control group （both P〈0.05）, which had no significant difference between the two latter groups （P〉0.05）; after exposure to HMGB1 for 4 h, the TNF-a level in HSF-1 overexpression group was significantly decreased compared with negative control group or blank control group （both P〈0.05）, which had no significant difference between the two latter groups （P〉0.05）; after exposure to HMGB 1 for different time periods, the expressions of MAPK pathway related proteins that included p-ERK, p-JNK and p-p38 as well as NF-kB pathway related protein p-IKα-B all showed no significant difference among the groups of cells （all P〉0.05）; results of EMSA showed that the expression of the gray-scale for NF-kB in HSF-1 overexpression group was significantly lower than that in negative control group or blank control group （both P〈0.05）, which had no significant difference between the two latter gr

关 键 词：炎症 热休克反应 高迁移率族蛋白质类 

分 类 号：R364.5[医药卫生—病理学]