异甘草酸镁上调血红素氧合酶-1对巨噬细胞过度分泌炎症因子的作用研究  

Magnesium isoglycyrrhetate inhibits the over - release of inflammatory mediators in LPS - stimulated macrophages through up - regulating heine oxygenation - 1

作　　者：张磊[1] 蒋磊[1] 张丽坤[1] 龚睿[1] 潘尚哈[1] 赵鸣雁[1] 

机构地区：[1]哈尔滨医科大学附属第一医院ICU,黑龙江哈尔滨150001

出　　处：《中国急救医学》2017年第11期983-987,共5页Chinese Journal of Critical Care Medicine

基　　金：黑龙江省自然科学基金（ZJY0704-02）;国家自然科学基金（81171785）;哈尔滨医科大学研究生创新科研项目（YJSCX2015-21HYD）

摘　　要：目的探讨异甘草酸镁（MgIG）在脂多糖（LPS）刺激的RAW264．7巨噬细胞中是否能通过上调血红素氧合酶-1（HO-1）起到抑制高迁移率族蛋白B1（HMGB1）和一氧化氮（NO）释放的作用。方法用LPS刺激小鼠巨噬细胞RAW264．7制作炎症细胞模型。①先用不同浓度的MgIG与LPS合并处理24h，分别为Control组、LPS组和LPS＋MglG不同剂量组（O．01mg／mL、0．10mg／mL、0．50mg／mL和1．00mg／mL）；②再将培养细胞随机分为不同处理组（每组n=5）：Control组、LPS组、LPS＋MdG（1．00mg／mL）和LPS＋MdG（1．00m∥mL）＋血红素氧化酶抑制剂[ZnPPIX（10μmol／L）]组。用ELISA方法测定上清HMGBl浓度，用Griess法测定上清液NO的浓度水平，用Western bolt检测细胞内HO-1和iNOS蛋白的表达。结果①与Control组比较，LPS能够明显增加上清液HMGB1和NO浓度（均P〈0．05），并且在一定程度上增加细胞内HO-1的蛋白表达水平（P〈0．05）；与LPS组比较，LPS＋MgIG不同剂量组均减少了HMGB1和NO的释放（均P〈0．05），并且随着剂量的增加，HMGB1和NO的上清液水平降低越明显，而HO-1的表达水平在LSP＋MgIG（0．10mg／mL、0．50mg／mL和1．00mg／mL）组均明显诱导增加（均P〈0．05）。②与Control比较，LPS同样明显增加上清液HMGB1和NO浓度（均P〈0．05），以及明显增加细胞内HO-1和可诱导一氧化氮合酶（iNOS）蛋白表达水平（均P〈0．05）；与LPS组比较，LPS＋MgIG（1．00mg／mL）组上清液中HMGBl和NO浓度明显下降（均P〈0．05），细胞内HO-1的蛋白水平明显增加（P〈0．05），细胞内iNOS的蛋白水平则明显下降（P〈0．05），而LPS＋MgIG（1．00mg／mL）＋ZnPPIX（10μmol／L）组上清液HMGB1和NO浓度、细胞内HO-1和iNOS的蛋白水平差异均无统计学意义（均P〉0．05）。结论MglG能抑制LPS刺激巨噬细胞过度释放HMGBl和NO，能明显抑制LPS诱导的iNOS表达，并且依赖于HObjective To investigate whether magnesium isoglycyrrhetate （MgIG） can inhibit high mobility group box - 1 protein （ HMGB1 ） and nitric oxide （NO） release by up - regulating heme oxygenase - 1 （ HO - 1 ） in lipopolysaccharide （LPS） - stimulated RAW264.7 macrophages. Methods RAW264.7 macrophages stimulated with LPS were used as inflammatory cell models. ①The cells were treated with LPS and different concentrations of MgIG for 24 hours firstly： the control group, LPS group and LPS ＋ MgIG groups （0.01 mg/mL,0.10 mg/mL,0.50 mg/mL and 1.00 mg/mL）. Secondly, the cells were randomly divided into the control group, LPS group, LPS ＋ MgIG （ 1.00 mg/mL） group and LPS ＋ MgIG （1.00 mg/mL） ＋ ZnPPlX （10 μmol/L） group. The level of HMGB1 in the supernatant was measured by ELISA. The concentration of NO in the supernatant was measured by Griess method. The expression of HO - 1 and iNOS protein was detected by Western blotting. Results ②Compared with the control group, LPS significantly increased the levels of HMGB1 and NO in the supernatant （P 〈 0.05）, and increased the expression of HO - 1 protein （P 〈 0.05 ）. Compared with the LPS group, the levels of HMGB1 and NO in the supernatant were both decreased with the increased dose of MglG （ P 〈 0.05 ）, while the expression of HO - 1 was significantly increased with the increased dose of MglG （ P 〈 0.05 ）. ②Compared with Control, LPS also siguificanfly increased the concentration of HMGB1 and NO in supernatant （ P 〈 0.05 ）, and notably increased HO - 1 and inducible nitric oxide synthase （iNOS） protein expression （P 〈0.05）. Compared with the LPS group, the levels of HMGB1 and NO in the supernatant and iNOS protein expression in the cell were all markedly decreased by MglG （1. 130 mg/mL） （P 〈0. 05）, while the HO- 1 protein expression in the cell was significantly augmented （P 〈 0.05）. However, no differences were detected between the LPS group and LPS ＋ MgIG （1.00 mg/mL）

关 键 词：异甘草酸镁(MgIG) 血红素氧合酶-1(HO-1) 高迁移率族蛋白B1(HMGB1) 一氧化氮(NO) 脓毒症 

分 类 号：R562.25[医药卫生—呼吸系统]