国际麻醉学与复苏杂志   2010, Issue (3): 230-232
    
异丙酚对脂多糖诱导的小鼠巨噬细胞表达释放高迁移率族蛋白1的抑制作用
郭远波 唐靖 杨艳琴 古妙宁1()
1.510515 广州， 南方医科大学附属南方医院麻醉科（郭远波、 唐靖、 杨艳琴、 古妙宁）
Inhibition of propofol on HMGB1 expression and releasing from mouse macrophage cells induced by lipopolysaccharide
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摘要：

目的：探讨异丙酚对脂多糖（lipopolysaccharide, LPS）诱导小鼠巨噬细胞RAW 264.7表达和释放高迁移率族蛋白1（high mobility group box 1 protein，HMGB1）的抑制作用。方法：检测LPS刺激后，传代培养的小鼠RAW 264.7细胞表达分泌HMGB1的时间梯度，确定RAW 264.7细胞表达分泌HMGB1浓度最高的时间点。随后将RAW 264.7细胞分3组接种于6孔板，每组为两孔：即仅加培养液的对照组（control组）；加250 μg/L LPS的诱导组（LPS组）；加250 μg/L LPS基础上加50 μmol/L异丙酚的干预组（LPS+异丙酚组）。培养至HMGB1表达分泌浓度最高的时间点后，收集细胞及细胞培养上清，用RT-PCR和Western blot法分别检测各组细胞内HMGB 1 mRNA表达水平和HMGB1蛋白表达水平，用Western blot法检测细胞培养上清中HMGB1含量的变化。结果：LPS刺激RAW 264.7细胞后16 h，HMGB1表达达到高峰，随后有所下降；加入异丙酚干预后，细胞HMGB1表达及释放显著低于LPS刺激组 ( P <0.05); LPS刺激RAW 264.7细胞后16 h，HMGB1 mRNA表达水平明显增强，而加入异丙酚干预后，则明显抑制HMGB1 mRNA表达水平，各组细胞中HMGB1蛋白含量变化和mRNA水平变化一致。结论 异丙酚抑制LPS诱导的RAW264.7细胞表达和释放HMGB1,可能对LPS血症具有一定保护作用。
关键词： 异丙酚；高迁移率族蛋白质1；脂多糖；RAW264. 7细胞
Abstract:

Objective To explore the inhibition effect of propofol on HMGB1 expression and releasing from mouse macrophage cell RAW264.7 induced by LPS. Methods Detect the time course of HMGB1 expression of RAW264.7 cells using RT-PCR and western blot. Then cells were divided into three groups: control,250 μg/L LPS stimulated,250 μg/L LPS stimulated simultaneously with 50 μmol/L propofol. HMGB1 expression and releasing were detected using RT-PCR and western blot when cell were stimulated until the time the HMGB1 expression got to the highest level. Results 16 h after stimulated by LPS, the HMGB1 expression got to the highest level. When LPS stimulated with 50 μmol/L propofol, the expression and releasing of HMGB1 by RAW264.7 were inhibited (P <0.05)；16 h after stimulated by LPS, the HMGB1 mRNA expression got to the highest level,When LPS stimulated with 50 μmol/L propofol, the expression and releasing of HMGB1 mRNA by RAW264.7 were inhibited significantly,in each group ,the changes of HMGB1 Protein are the same to that of HMGB1 mRNA. Conclusion Propofol can inhibit the expression and releasing of HMGB1 by RAW264.7, which may be play a role in protect of sepsis by LPS.
Key words: Propofol; High mobility group proteins; Lipopolysaccharide; RAW 264.7 cells