【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（H2O2）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 H₂O₂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。更多还原

【Abstract】 【Objective】To investigate the role of PI3 K/Akt/eNOS signaling pathway in the protective effect of hydrogen sulfide(H2 S)on endothelial cell senescence induced by hydrogen peroxide(H2 O2).【Methods】Senescence in human umbilical vein endothelial cells(HUVEC)was induced by 60 μmol/L H2 O2 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 60 μmol/L H2 O2,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 H2 O2,and this may be highly associated with PI3 K/Akt/eNOS signaling pathway.更多还原