外源性硫化氢通过PI3K/Akt/eNOS通路改善过氧化氢诱导的人脐静脉内皮细胞衰老  

Exogenous Hydrogen Sulfide Improves Hydrogen Peroxide-Induced Senescence of Human Umbilical Vein Endothelial Cells via PI3K/Akt/eNOS Pathway

作　　者：牛海名 李建伟[1] 陈妙莲[1] 刘定辉 吴琳[2] 凌叶盛 余显冠[2] 钱孝贤 NIU Hai-ming;LI Jian-wei;CHEN Miao-lian;LIU Ding-hui;WU Lin;LING Ye-sheng;YU Xian-guan;QIAN Xiao-xian(Department of Critical Care,Zhongshan People’s Hospital/Zhongshan Affiliated Hospital,Sun Yat-sen University,Zhongshan 528400,China;Department of Cardiology,The Third Affiliated Hospital,Sun Yat-sen University,Guangzhou 510630,China;Institute for Integrated Traditional Chinese and Western Medicine,Sun Yat-sen University,Guangzhou 510630,China)

机构地区：[1]中山市人民医院/中山大学附属中山医院重症医学科,广东中山528400 [2]中山大学附属第三医院心血管内科,广东广州510630 [3]中山大学中西医结合研究所,广东广州510630

出　　处：《中山大学学报（医学科学版）》2021年第4期535-542,共8页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：国家自然基金(81370447);广东省省科技计划项目(2016A050502014);中山市社会公益科技基金(2019B1053)。

摘　　要：【目的】本研究拟探讨磷脂酰肌醇3激酶/蛋白激酶B/内皮型一氧化氮合酶(PI3K/Akt/eNOS)通路对硫化氢(H2S)改善内皮细胞衰老的作用。【方法】建立60μmol/L过氧化氢(H_(2)O_(2))诱导的人脐静脉内皮细胞(HUVEC)衰老模型;衰老模型的评估及不同浓度硫氢化钠(NaHS)预处理对衰老的作用均通过检测纤溶酶原激活物抑制剂1(PAI-1)的表达水平和衰老相关β-半乳糖苷酶(SA-β-Gal)染色阳性率来研究评估。同时通过Western blot法检测各组内皮细胞中蛋白激酶B(Akt)蛋白、磷酸化蛋白激酶B(p-Akt)蛋白和内皮型一氧化氮合酶(eNOS)蛋白的表达,通过硝酸盐还原酶法检测一氧化氮(NO)含量。【结果】与对照组相比,HUVEC经60μmol/L H_(2)O_(2)预处理后,能显著提高细胞SA-β-gal阳性率和PAI-1的表达。而NaHS的预处理可以显著减少PAI-1的表达和细胞SA-β-gal阳性率,但可增加p-Akt蛋白、eNOS蛋白的表达及增加NO含量。【结论】NaHS可以改善H2O2诱导的内皮细胞衰老,其机制与PI3K/Akt/eNOS通路有关。【Objective】To investigate the role of PI3K/Akt/eNOS signaling pathway in the protective effect of hydrogen sulfide(H2S)on endothelial cell senescence induced by hydrogen peroxide(H2O2)【.Methods】Senescence in human umbilical vein endothelial cells(HUVEC)was induced by 60μmol/L H_(2)O_(2) for 1 h,and the effects of NaHS on the senescence were examined by the expression of plasminogen activator inhibitor 1(PAI-1)and senescence-associated β-galactosidase(SA-β-Gal)staining.Meanwhile,the expression of protein kinase B(Akt),Phospho protein kinase B(p-Akt),and endothelial nitric oxide synthase(eNOS)were examined by western blot,and the senescence-related parameters.The content of the nitric oxide(NO)was measured by nitrate reduction method.【Results】Compared with the control group,after being treated with 60μmol/L H_(2)O_(2),the number of SA-β-gal positive cells and the expression of PAI-1 were significantly increased.However,after pretreatment of NaHS,the expression of PAI-1,the ratio of SA-β-gal positive cells was significantly decreased,but the expression of p-Akt,eNOS and the NO concentration increased.【Conclusion】NaHS could reverse HUVEC senescence induced by H2O2,and this may be highly associated with PI3K/Akt/eNOS signaling pathway.

关 键 词：硫化氢 人脐内皮细胞 过氧化氢 衰老 PI3K/Akt/eNOS通路 

分 类 号：R339.38[医药卫生—人体生理学]