雷帕霉素抑制LPS诱导的RAW264．7细胞表达和释放HMGB1的机制  

Rapamycin inhibits HMGB1 expression and releases in RAW264.7 cells induced by lipopolysaccharides in vitro

作　　者：赵中夫[1] 韩德五[2] 张芸[1] 刘明社[1] 张国英[1] 

机构地区：[1]长治医学院肝病研究所,山西长治046000 [2]山西医科大学肝病研究所,太原030001

出　　处：《中国病理生理杂志》2008年第7期1418-1421,共4页Chinese Journal of Pathophysiology

基　　金：山西省自然科学基金资助项目(No.2008011073-4)

摘　　要：目的:探讨雷帕霉素抑制LPS诱导小鼠RAW264.7细胞表达和释放HMGB1的作用机制。方法:传代培养的小鼠RAW264.7细胞分5组接种于6孔板,即仅加培养液的对照组;加250μg/LLPS的诱导组;诱导组基础上加100μg/L雷帕霉素的干预组;干预组基础上加rTNF-α50μg/L的抗干预组;及抗干预组基础上加抗鼠TNF-α100μg/L的抗体中和组。培养4h后,ELISA法检测对照组、诱导组和干预组上清液中TNF-α水平;培养24h后,RT-PCR和Western blotting法分别检测各组细胞内HMGB1 mRNA表达水平和上清液中HMGB1含量。结果:培养4h,干预组上清液中TNF-α水平与对照组比无显著差异(P>0.05),但明显低于诱导组(P<0.05);培养24h,与对照组比,诱导组细胞内HMGB1 mRNA表达明显增强(P<0.05),上清液中HMGB1含量也明显增多(P<0.05);干预组明显减少了HMGB1mRNA表达及上清液中HMGB1含量(P<0.05);与干预组比,抗干预组细胞HMGB1 mRNA表达及上清液中HMGB1含量明显增加(P<0.05);抗体中和组细胞HMGB1 mRNA表达水平及上清液中HMGB1含量和干预组无显著差异(P>0.05)。结论:雷帕霉素抑制LPS诱导RAW264.7细胞表达和释放HMGB1,可能部分地与其抑制TNF-α的表达有关。AIM： To observe the mechanism that rapamycin （RPM） affects HMGB1 expression and release in RAW264.7 cells induced by lipopolysaccharides （LPS）. METHODS ： RAW264.7 cells were cultured in six wells plate and divided into five groups： control group, 250 μg/L LPS treatment group, 100 μg/L RPM treatment group, 50 μg/L rT NF -α treatment group and 100 μg/L TNF-α antibody treatment group. After 4 h treatment, the TNF -α level in the culture media was evaluated by ELISA assay. After 24 h, the expression of HMGB1 mRNA was measured by RT - PCR, and HMGBI protein level in the culture media was determined by Western blotting analysis. RESULTS ： TNF-α level in the culture media of RAW264.7 cells has no significant difference between RPM treatment group and control group （P 〉 0. 05 ）. Both HMGB1 mRNA expression and HMGB1 protein level were remarkably higher in LPS treatment group than that in control group （P〈0.05）. RPM attenuated LPS -induced HMGB1 mRNA and HMGB1 accumulation. Compared with that in RPM treatment group, HMGB1 accumulation was increased in rTNF -α treatment group, and had no significant difference in TNF -α antibody treatment group （ P 〉 0. 05 ）. CONCLUSION： RPM inhibits HMGB1 expression not only by directly suppressing STAT3 activation, but also by indirectly reducing TNF -α level.

关 键 词：雷帕霉素 高迁移率族蛋白质类 RAW264.7细胞 肿瘤坏死因子 

分 类 号：R363[医药卫生—病理学]