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低镁对脂多糖诱导巨噬细胞表达和释放HMGB1的影响
Low magnesium promotes 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,江继宏,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疼痛科,上海交通大学附属第一人民医院麻醉科
Department of Pain,Shanghai First People’s Hospital, Shanghai Jiaotong University;Department of Anesthesiology,Shanghai First People’s Hospital, Shanghai Jiaotong University
关键词:
低镁;硫酸镁;高迁移率族蛋白质1;脂多糖;RAW264.7细胞
low magnesium;Magnesium Sulphate;High mobility group proteins1;Lipopolysaccharide;RAW 264.7 cell
摘要:
目的 探讨低镁对脂多糖(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)的促进作用。方法 传代培养的小鼠RAW 264.7细胞分4组接种于6孔板,每组为三孔:含1mmol/L硫酸镁的RPMI1640培养基组(C1组);含0.1mmol/L硫酸镁的RPMI1640培养基组(C0.1组);含1mmol/L硫酸镁的RPMI1640培养基加500ng/ml LPS的刺激组(L1组);含0.1mmol/L硫酸镁的RPMI1640培养基加500ng/ml LPS的刺激组(L0.1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;C0.1组与C1组相比,细胞培养上清中HMGB1释放差异无统计学意义(P>0.05);L0.1组与L1组相比,细胞培养上清中HMGB1释放显著升高(P<0.05);LPS刺激RAW 264.7细胞24h后,细胞内HMGB1 mRNA表达水平明显增强;C0.1组与C1组相比,细胞内HMGB1 mRNA表达水平差异无统计学意义(P>0.05);L0.1组与L1组相比,细胞内HMGB1 mRNA表达水平显著升高(P<0.05)。结论 低镁促进LPS诱导的RAW264.7细胞表达和释放HMGB1,通过促进与脓毒症致死性密切相关的关键炎症因子释放,可能对脓毒症患者的预后有一定损害作用。
0bjective To explore the promoted effect of low magnesium 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 4 groups, each with three holes:RPMI1640 Medium containing 1mmol/L magnesium sulphate(C1 group);RPMI1640 Medium containing 0.1mmol/L magnesium sulphate(C0.1 group); RPMI1640 Medium containing 1mmol/L magnesium sulphate +LPS treatment group (L1 group); RPMI1640 Medium containing 0.1mmol/L magnesium sulphate +LPS treatment group(L0.1 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 L1 group and L0.1 group was significantly higher than that in C1 group and C0.1 group(P<0.05); Compared with C1 group ,the HMGB1 expression of C0.1 group have no significant difference(P>0.05); the cell culture supernatant level of HMGB1 in L0.1 group was significantly higher than that in L1 group(P<0.05); 24h after stimulated by LPS,the HMGB1 mRNA expression in L1 group and L0.1 group was significantly higher than that in C1 group and C0.1 group(P<0.05);Compared with C1 group, the HMGB1 mRNA expression of C0.1 group have no significant difference(P>0.05);The HMGB1 mRNA expression in L0.1 group were significantly higher than that in L1 group (P<0.05). Conclusion Low magnesium promotes the expression and releasing of HMGB1 by RAW264.7,which may be play a harmfull effect in sepsis .
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