硫氢化钠增加高糖高脂条件下小鼠心房肌细胞系HL-1谷胱甘肽合成  

Sodium hydrosulfide increases glutathione synthesis in cardiac muscle cell line HL-1 under hyperglycemia and hyperlipidemia

作　　者：张伟才[1] 刘肆仁[1] 王尚农[2] ZHANG Weicai;LIU Siren;WANG Shangnong(Department of Cardiology,Beijing No.6 Hospital,Beijing 100007,China;Department of Endocrinology,Beijing No.6 Hospital,Beijing 100007,China)

机构地区：[1]北京市第六医院心内科,北京100007 [2]北京市第六医院内分泌科,北京100007

出　　处：《基础医学与临床》2024年第1期69-76,共8页Basic and Clinical Medicine

摘　　要：目的研究硫氢化钠(NaHS)是否通过调节谷胱甘肽(GSH)的合成,降低活性氧自由基(ROS)产生,改善小鼠2型糖尿病心肌病(DCM)。方法将小鼠心房肌细胞系HL-1分为对照组、高葡萄糖(HG:40 mmol/L)和棕榈酸(Pal:500μmol/L)处理组;以及硫氢化钠(NaHS,100μmol/L)、DL-炔丙基甘氨酸[PPG,胱硫醚γ裂解酶(CSE)抑制剂,1 mmol/L]和N-乙酰-L-半胱氨酸(NAC,ROS抑制剂,5 mmol/L)处理72 h组。Western blot检测CSE和谷胱甘肽合成酶(GSS)的表达;二氢乙锭(DHE)和二氯氟甲烷(DCFH)检测ROS含量;免疫共沉淀检测核因子红细胞系2相关因子2(Nrf2)泛素化水平及Nrf2与肌肉特异性环指蛋白1(Murf1)的相互作用。结果与对照组比高糖高脂处理HL-1细胞后CSE、溶质载体家族7成员11(SLC7A11)、谷氨酸半胱氨酸连接酶催化亚基C(GCLC)、谷氨酸半胱氨酸连接酶修饰亚基M(GCLM)、谷胱甘肽合成酶(GSS)的表达水平下降,而NaHS能恢复其表达。高糖高脂组ROS含量高于NaHS组。与NaSH组比高糖高脂条件下Murf1与Nrf2的相互作用增加,Nrf2泛素化水平明显增加。结论硫氢化钠减轻Nrf2的泛素化,增加GSH合成关键酶的表达。Objective To investigate sodium hydrogen sulfide(NaHS)with function of regulating glutathione(GSH)synthesis to reduce reactive oxygen species(ROS)production in type 2 diabetic cardiomyopathy(DCM).Methods Mouse cardiomyocyte cell line HL-1 was incubated with high concentration of glucose(HG:40 mmol/L)and palmitate(Pal:500μmol/L)as a cell model of type 2 DCM.HL-1 cells were incubated with NaHS(100μmol/L),DL-propargylglycin(PPG,1 mmol/L)and N-acetyl-l-cysteine(NAC,5 mmol/L),respectively for 72 hours.The expression of cystathionine-γ-lyase(CSE)and the key enzymes of glutathione production was tested by Western blot.Dihydroethidium(DHE)and dichlorofluoromethane(DCFH)were used to detect the content of ROS in HL-1 cells.Cell viability was detected by CCK8 kit.The content of total GSH was detected.The interaction between muscle specific ring finger protein 1(Murf1)and nuclear factor erythroid-derived 2-related factor 2(Nrf2)and Nrf2 ubiquitylation was determined by co-immunoprecipitation(co-IP).Results Compared with control group,the expression level of CSE,solute carrier family 7 members 11(SLC7A11),glutamate cysteine ligase C(GCLC),glutamate cysteine ligase M(GCLM)and glutathione synthetase(GSS)in HL-1 cells treated incubated with high glucose and palmitate was decreased,however,NaHS was found to restore it.NaHS reduced the content of ROS in HL-1 cells treated with high glucose and palmitate.The interaction between murf1 and Nrf2 was confirmed by co-immunoprecipitation(Co-IP).Compared with NaHS group,the ubiquitylation level of Nrf2 was enhanced in high glucose and palmitate group.Conclusions Sodium hydrosulfide may reduce the ubiquitylation level of Nrf2 and promote the expression of key enzymes of GSH synthesis.

关 键 词：糖尿病心肌病 硫化氢 谷胱甘肽 核因子红细胞系2相关因子2(Nrf2) E3连接酶肌环指蛋白-1(Murf1) 

分 类 号：R542.2[医药卫生—心血管疾病] R587.2[医药卫生—内科学]