外源性硫化氢对脓毒症巨噬细胞氧化还原平衡的影响  

Effects of exogenous hydrogen sulfide on redox equilibrium of septic macrophages

作　　者：王兴程 杨磊[1] 王斌生[1] 彭超[1] 马立斌[1] 柴深 Wang Xing-cheng;Yang Lei;Wang Bin-sheng;Peng Chao;Ma Li-bin;Chai Chen(Department of General Surgery,First Affiliated Hospital of Lanzhou University,Lanzhou 730000,China)

机构地区：[1]兰州大学第一医院普外三科,甘肃兰州730000

出　　处：《中国急救医学》2019年第1期77-81,共5页Chinese Journal of Critical Care Medicine

基　　金：兰州市科技局项目(2015-2-65).

摘　　要：目的研究外源性硫化氢对脓毒症巨噬细胞氧化还原平衡的影响。方法培养小鼠腹腔巨噬细胞株J774,分为缓冲液(PBS)(1 mL/mL)对照组、脂多糖(LPS)(1μg/mL)组、炔丙基甘氨酸(PAG)(1 mmol/L)+LPS(1μg/mL)组、硫化氢缓释供体(GYY4137)(500μmol/L)组、GYY4137(500μmol/L)+LPS(1μg/mL)组,酶联免疫吸附(ELISA)法测定各组细胞培养上清中活性氧(ROS)、丙二醛(MDA)、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GSH-Px),电子显微镜观察细胞内线粒体形态的变化。结果最终测得LPS组细胞上清中ROS为(90.293±0.217)U/mL,MDA为(1.665±0.068)nmol/mL,SOD为(89.857±3.00)U/mL,CAT为(8.099±0.130)U/mL,GSH-Px为(10.287±0.335)μmol/L。GYY4137+LPS组细胞上清中ROS平均浓度为(25.819±2.988)U/mL,MDA为(0.355±0.065)nmol/mL,SOD为(189.690±0.290)U/mL,CAT为(189.690±0.290)U/mL,GSH-Px为(287.235±0.864)μmol/L。与LPS组比较,GYY4137+LPS组氧化应激产物ROS及氧化应激标志物MDA水平较低(P﹤0.05),抗氧化剂SOD、CAT、GSH-Px水平较高(P﹤0.05),细胞及线粒体损伤程度也较小。结论硫化氢对脓毒症巨噬细胞氧化还原平衡有一定的保护作用,这种保护作用的机制可能是硫化氢减轻了脓毒症时线粒体的氧化应激损伤。Objective To explore the effects of exogenous hydrogen sulfide(H2S)on redox balance of macrophages in sepsis.Methods The macrophage strain J774 was divided into phosphate buffer saline(PBS)(1 mL/mL)control group,lipopolysaccharide(LPS)(1μg/mL)group,DL-propargylglycine(PAG)(1 mmol/L)+LPS(1μg/mL)group,GYY4137(500 jimol/L)group and GYY4137(500μmol/L)+LPS(1μg/mL)group.In the supernatant of cell culture of each group,the levels of reactive oxygen species(ROS),malondialdehyde(MDA),superoxide dismutase(SOD),catalase(CAT)and glutathione peroxidase(GSH-Px)were measured by ELISA.The changes of mitochondrial morphology in cells were observed by electron microscope.Results Finally,the average concentration of ROS in supernatant of LPS group was(90.293±0.217)U/mL,MDA was(1.665±0.068)nmo/mL,SOD was(89.857±3.002)U/mL,CAT was(8.099±0.130)U/mL,GSH-Px was(10.287±0.335)jimol/L.In the supernatant of GYY4137+LPS group,the average concentration of ROS was(25.819±2.988)U/mL,MDA was(0.355±0.065)nmol/mL,SOD was(189.690±0.290)U/mL,CAT was(189.690±0.290)U/mL,GSH-Px was(287.235±0.864)pnol/L.The levels of ROS and MDA in the GYY4137+LPS group were significantly lower than those in the LPS group with significant difference(P﹤0.05).The levels of antioxidant SOD,CAT andGSH-Px were considerably higher with significant difference(P﹤0.05),and the degree of damage to cells and mitochondria was also lower.Conclusion Hydrogen sulfide has a protective effect on the redox balance of septic macrophages,which may be due to its direct inhibition of mitochondrial oxidative stress.

关 键 词：硫化氢 脓毒症 巨噬细胞 氧化应激 

分 类 号：R459.7[医药卫生—急诊医学]