外源性硫化氢改善人脐静脉内皮细胞早熟性衰老与eNOS表达的关系

目的拟探讨硫化氢(H₂S)延缓人脐静脉内皮细胞(HUVECs)衰老与内皮型一氧化氮合酶(eNOS)的关系,为延缓HUVECs衰老提供新的靶点。方法建立60μmol/L过氧化氢(H₂O₂)1 h诱导HUVECs衰老模型;通过检测衰老相关β-半乳糖苷酶(SAβ-Gal)染色阳性率来研究评估外源性硫氢化钠(NaHS)预处理对衰老的作用。同时通过Western blot法和RT-PCR法检测内皮细胞中eNOS蛋白和eNOS mRNA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,HUVECs经60μmol/L H₂O₂预处理后,能显著提高细胞SAβ-Gal阳性率。而NaHS的预处理可以显著减少细胞SAβ-Gal阳性率,但可增加eNOS蛋白、eNOS mRNA的表达及增加NO含量。结论NaHS可以通过上调内皮细胞中eNOS的活性和蛋白表达改善H₂O₂诱导的内皮细胞衰老。

Effect of exogenous hydrogen sulfide on premature senescence of human umbilical vein endothelial cells and its relationship with eNOS expression

Objective This study aims to explore the relationship between hydrogen sulfide(H₂S)delaying the aging of human umbilical vein endothelial cells(HUVECs)and endothelial nitric oxide synthase(e NOS),so as to provide a new target for delaying the aging of HUVECs.Methods Senescence in human umbilical vein endothelial cells(HUVECs)was induced by 60μmol/L H₂O₂ for 1 h,and the effects of NaHS on the senescence were examined by senescence-associatedβ-galactosidase(SAβ-Gal)staining.Mean while,the expression of endothelial nitric oxide synthase(e NOS)were examined by western blot,Real time PCR was used to detect the expression of eNOS mRNA.The content of the nitric oxide(NO)was measured by nitrate reduction method.Results Compared with the control group,after treated with 60μmol/L H₂O₂,the number of SAβ-Gal positive cells were significantly increased.However,after pretreatment of NaHS,the ratio of SAβ-Gal positive cells was significantly decreased,but the expression of eNOS,eNOS mRNA and the NO concentration increased.Conclusion Na HS can improve H₂O₂ induced endothelial cell aging by up regulating the activity and protein expression of eNOS in endothelial cells.