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硫酸镁对LPS诱导巨噬细胞表达和释放HMGB1的影响
Effects of magnesium sulphate on the expression and release of HMGB1 in macrophages induced by lipopolysaccharide

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DOI:
作者:
刘朝辉，张俊杰，黄小静，黄丽娜，李士通，汪正平
LIU Zhaohui，ZHANG Junjie，LI Shitong，WANG Zhengping
作者单位:
上海交通大学附属第一人民医院麻醉科，上海交通大学附属第一人民医院疼痛科
Department of Anesthesiology，Shanghai First People’s Hospital,Shanghai Jiaotong University
关键词:
硫酸镁；高迁移率族蛋白质1；脂多糖；RAW264.7细胞
Magnesium Sulphate；High mobility group proteins1；Lipopolysaccharide；RAW 264.7 cell
摘要:
【摘要】 目的 探讨硫酸镁对脂多糖(1ipopolysaccharide，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 传代培养的小鼠RAW 264.7细胞分5组接种于6孔板，每组为三孔：即仅加RPMI1640培养液的对照组(C组)；在RPMI1640培养液的基础上加500ng/ml LPS的诱导组(LPS组)；在诱导组基础上加1 mmol/L硫酸镁的干预组(M1组)；加5 mmol/L硫酸镁的干预组(M5组)；加10 mmol/L硫酸镁的干预组(M10组)。孵育24h后，收集细胞及细胞培养上清，用RT-PCR和ELISA法分别检测各组细胞内HMGB1 mRNA表达水平和细胞培养上清中HMGB1蛋白含量的变化。结果 LPS刺激RAW 264.7细胞24h后，细胞培养上清中HMGB1释放显著升高；加硫酸镁干预后，M5和M10组细胞HMGB1释放显著低于LPS诱导组 (P<0.05)； LPS刺激RAW264.7细胞24h后，细胞内HMGB1 mRNA表达水平明显增强；而加入硫酸镁干预后，M5和M10组细胞HMGB1 mRNA表达水平显著低于LPS诱导组（P<0.05）。结论 硫酸镁抑制LPS诱导的RAW264.7细胞表达和释放HMGB1，通过抑制与脓毒症致死性密切相关的关键炎症因子，可能对脓毒症患者具有一定保护作用。
【Abstract】0bjective To explore the effects of magnesium sulphate on the expression and release of the high mobility group box-1(HMGB1) in mouse macrophage cell RAW264.7 induced by lipopolysaccharide(LPS)．Methods Mouse macrophage cell RAW264.7 incubated in 6-well tissue culture plates were divided into 5 groups, each with three holes：the control group; LPS group; magnesium sulphate 1mmol/L＋ LPS treatment group(M1 group); magnesium sulphate 5mmol/L＋ LPS treatment group(M5 group);magnesium sulphate 10mmol/L＋ LPS treatment group(M10 group). 24h after stimulated by LPS, the HMGB1 expression and releasing were detected using RT-PCR and ELISA. Results 24h after stimulated by LPS，the cell culture supernatant level of HMGB1 in LPS group was significantly higher than that in the control group(P<0.05); the cell culture supernatant level of HMGB1 in M5 and M10 group was significantly lower than that in LPS group(P<0.05);24h after stimulated by LPS，the HMGB1 mRNA expression in LPS group was significantly higher than that in the control group(P<0.05);The HMGB1 mRNA expression in M5 and M10 group were significantly lower than that in LPS group (P<0.05). Conclusion Magnesium sulphate can inhibit the expression and releasing of HMGB1 by RAW264.7，which may be play a role in protect of sepsis by LPS.

 

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