硫化氢对氧化型低密度脂蛋白诱导的人脐静脉内皮细胞凋亡的影响
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国家自然科学基金项目(30971169);;衡阳市科学技术发展计划项目(2009KJ10)


Investigation on the Effect and Mechanisms of Hydrogen Sulfide Against Apoptosis Induced by ox-LDL in Human Umbilical Vein Endothelial Cells
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    摘要:

    目的探讨硫化氢对氧化型低密度脂蛋白诱导的人脐静脉内皮细胞凋亡的影响及有关机制。方法人脐静脉内皮细胞分别经不同浓度(25、50、100、200μmol/L)和不同时间(6、12、24 h)硫氢化钠预孵育24 h后加入100 mg/L氧化型低密度脂蛋白处理24 h,Hoechst 33258荧光染色和流式细胞术检测细胞凋亡,罗丹明123染色检测细胞线粒体膜电位变化,双氢罗丹明123染色检测细胞内活性氧的含量变化。结果硫氢化钠预处理能以时间和浓度依赖性的方式抑制氧化型低密度脂蛋白诱导的人脐静脉内皮细胞凋亡(均P<0.01),硫氢化钠或N-乙酰半胱氨酸预先处理能显著阻断氧化型低密度脂蛋白引起的人脐静脉内皮细胞线粒体膜电位降低、活性氧生成增多(均P<0.05)。结论硫化氢能显著抑制氧化型低密度脂蛋白诱导的人脐静脉内皮细胞凋亡,其机制与减少细胞线粒体膜电位及降低活性氧生成有关。

    Abstract:

    Aim To investigate the effects of hydrogen sulfide on ox-LDL-induced apoptosis in human umbilical vein endothelial cells(HUVEC) and the mechanisms involved.Methods The HUVEC were incubated with ox-LDL for 24 hours after treatment with NaHS at different concentrations(25,50,100,200 μmol/L) for 24 hours or for different incubation time(6,12,24 hours) at the concentration of 50 μmol/L.The apoptotic alterations of HUVEC were observed by Hoechst 33258 staining and flow cytometry(FCM),respectively.The change of mitochondrial membrane potential(MMP) and intracellular reactive oxygen species(ROS) generation were detected by Dihydrohodamine123(DHR123) staining and rhodamine123(Rh123) staining.Results Pre-treatment with NaHS remarkably inhibited apoptosis induced by ox-LDL in HUVEC in a time-and concentration-dependent manner(all P<0.01).The decrease of MMP and increase of the intracellular ROS generation in HUVEC induced by ox-LDL were significantly prohibited by treatment with NaHS or N-acetyl-L-cysteine(NAC)(all P<0.05).Conclusions H2S inhibits apoptosis induced by ox-LDL in HUVEC which is involved in preservation of mitochondrial membrane potential and attenuation of the intracellular ROS generation.

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任重,赵战芝,彭湘萍,谢巍,刘艳文,索荣,姜志胜.硫化氢对氧化型低密度脂蛋白诱导的人脐静脉内皮细胞凋亡的影响[J].中国动脉硬化杂志,2011,19(11):891~896.

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  • 收稿日期:2011-03-15
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