醛固酮诱导的氧化应激损伤内皮祖细胞介导的血管内皮修复功能
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摘要
【目的】进一步探索醛固酮对循环中内皮祖细胞(EPC)功能的影响及其分子机制有待。【方法】抽取健康志愿者外周血分离培养EPC,研究浓度梯度醛固酮干预(0,10,100,1000 nmol/L)对EPC体外迁移、黏附功能的影响。建立裸鼠颈动脉内皮拉脱损伤模型,在体研究醛固酮干预对EPC介导的血管内皮修复作用的影响。应用二氯二氢荧光素二醋酸酯(DCF)荧光探针研究醛固酮干预对EPC内活性氧簇(ROS)生成的影响。分别采用盐皮质激素受体(MR)拮抗剂—螺内酯、NADPH氧化酶阻断剂—夹竹桃麻素阻断EPC的MR和NADPH氧化酶,研究醛固酮诱导氧化应激损伤EPC血管内皮修复功能的分子信号通路。【结果】醛固酮干预明显抑制EPC体外迁移、黏附功能,损伤其介导的在体血管内皮修复能力。醛固酮干预导致EPC的ROS生成明显升高。螺内酯、夹竹桃麻素干预可以减少醛固酮诱导的ROS生成,并且可以对抗醛固酮损伤EPC功能。【结论】醛固酮通过MR激活NADPH氧化酶诱导氧化应激损伤EPC介导的血管内皮修复功能。
【Objective】To determine the effect and molecular mechanisms of aldosterone on the endothelial repair capacity of endothelial progenitor cells( EPC) in peripheral blood.【Method】EPC were cultured from peripheral blood mononuclear cells of healthy subjects. In vitro EPC function was assayed by migration and adhesion after treatment with different concentration of aldosterone(0, 10, 100,1000 nmol/L).In vivo endothelial repair capacity of EPC was evaluated by transplantation into a nude mouse carotid endothelial denudation model. The reactive oxygen species(ROS) was detected by 2',7 '-dichlorodihydrofluorescein diacetate( DCF)-fluorescent probe under fluorescence microscopy.【Result】Both the in vitro function and in vivo endothelial repair capacity of EPC were impaired obviously by aldosterone treatment. ROS production was distinctly increased after aldosterone treatment as well. Co-treatment with mineralocorticoid receptor inhibitor spironolactone or NADPH oxidase blocker apocynin prevented aldosterone-induced ROS production and recovered aldosterone-impaired EPC function.【Conclusion】Aldosterone-induced oxidative stress impairs endothelial repair capacity of EPC via MR-dependent activation of NADPH oxidase.
【Objective】To determine the effect and molecular mechanisms of aldosterone on the endothelial repair capacity of endothelial progenitor cells( EPC) in peripheral blood.【Method】EPC were cultured from peripheral blood mononuclear cells of healthy subjects. In vitro EPC function was assayed by migration and adhesion after treatment with different concentration of aldosterone(0, 10, 100,1000 nmol/L).In vivo endothelial repair capacity of EPC was evaluated by transplantation into a nude mouse carotid endothelial denudation model. The reactive oxygen species(ROS) was detected by 2',7 '-dichlorodihydrofluorescein diacetate( DCF)-fluorescent probe under fluorescence microscopy.【Result】Both the in vitro function and in vivo endothelial repair capacity of EPC were impaired obviously by aldosterone treatment. ROS production was distinctly increased after aldosterone treatment as well. Co-treatment with mineralocorticoid receptor inhibitor spironolactone or NADPH oxidase blocker apocynin prevented aldosterone-induced ROS production and recovered aldosterone-impaired EPC function.【Conclusion】Aldosterone-induced oxidative stress impairs endothelial repair capacity of EPC via MR-dependent activation of NADPH oxidase.
引文
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[14]丁美琳,贾敏,陈龙,等.高醛固酮血症对血管内皮功能及动脉弹性的影响[J].中华高血压杂志,2017,25(11):1275-1286.Ding ML,Jia M,Chen L,et al.The Effect of Hyperaldosteronemia on Endothelial Function and Arterial Elasticity[J].Chin J Hypertens,2017,25(11):1275-1286.
[15]Madamanchi NR,Vendrov A,Runge MS.Oxidative stress and vascular disease[J].Arterioscler Thromb Vasc Biol,2005,25(1):29-38.
[16]Bauersachs J,Fraccarollo D.Endothelial NO synthase target of aldosterone[J].Hypertension,2006,48(1):165-171.
[17]Galley JC,Straub AC.Redox Control of Vascular Function[J].Arterioscler Thromb Vasc Biol,2017,37(12):e178-e184.
[18]F?rstermann U,Xia N,Li H.Roles of Vascular Oxidative Stress and Nitric Oxide in the Pathogenesis of Atherosclerosis[J].Circ Res,2017,120(4):713-735.
[19]Chen L,Wu F,Xia WH,et al.CXCR4 gene transfer contributes to in vivo reendothelialization capacity of endothelial progenitor cells[J].Cardiovasc Res,2010,88(3):462-470.
[20]Wang JM,Yang Z,Xu MG,et al.Berberine-induced decline in circulating CD31+/CD42-microparticles is associated with improvement of endothelial function in humans[J].Eur J Pharmacol,2009,614(1-3):77-83.
[21]Milliez P,Girerd X,Plouin PF,et al.Evidence for an increased rate of cardiovascular events in patients with primary aldosteronism[J].J Am Coll Cardiol,2005,45(8):1243-1248.
[2]Lamping K.Endothelial progenitor cells sowing the seeds for vascular repair[J].Circ Res,2007,100(9):1243-1245.
[3]Zampetaki A,Kirton JP,Xu Q.Vascular repair by endothelial progenitor cells[J].Cardiovasc Res,2008,78(3):413-421.
[4]Leone AM,Valgimigli M,Giannico MB,et al.From bone marrow to the arterial wall:the ongoing tale of endothelial progenitor cells[J].Eur Heart J,2009,30(8):890-899.
[5]Werner N,Nickenig G.Influence of cardiovascular risk factors on endothelial progenitor cells:limitations for therapy[J]?Arterioscler Thromb Vasc Biol,2006,26(2):257-266.
[6]Sorrentino SA,Bahlmann FH,Besler C,et al.Oxidant stress impairs in vivo reendothelialization capacity of endothelial progenitor cells from patients with type 2 diabetes mellitus:restoration by the peroxisome proliferator-activated receptor-γagonist rosiglitazone[J].Circulation,2007,116(2):163-173.
[7]Giannotti G,Doerries C,Mocharla PS,et al.Impaired endothelial repair capacity of early endothelial progenitor cells in prehypertension:relation to endothelial dysfunction[J].Hypertension,2010,55(6):1389-1397.
[8]Xia WH,Yang Z,Xu SY,et al.Age-related decline in reendothelialization capacity of human endothelial progenitor cells is restored by shear stress[J].Hypertension,2012,59(6):1225-1231.
[9]Zhang XY,Su C,Cao Z,et al.CXCR7 upregulation is required for early endothelial progenitor cellmediated endothelial repair in patients with hypertension[J].Hypertension,2014,63:383-389.
[10]Schiffrin EL.Effects of aldosterone on the vasculature[J].Hypertension,2006,47(3):312-318.
[11]Nagata D,Takahashi M,Sawai K,et al.Molecular mechanism of the inhibitory effect of aldosterone on endothelial NO synthase activity[J].Hypertension,2006,48:165-171.
[12]Marumo T,Uchimura H,Hayashi M,et al.Aldosterone impairs bone marrow-derived progenitor cell formation[J].Hypertension,2006,48(3):490-496.
[13]Thum T,Schmitter K,Fleissner F,et al.Impairment of endothelial progenitor cell function and vascularization capacity by aldosterone in mice and humans[J].Eur Heart J,2011,32(10):1275-1286.
[14]丁美琳,贾敏,陈龙,等.高醛固酮血症对血管内皮功能及动脉弹性的影响[J].中华高血压杂志,2017,25(11):1275-1286.Ding ML,Jia M,Chen L,et al.The Effect of Hyperaldosteronemia on Endothelial Function and Arterial Elasticity[J].Chin J Hypertens,2017,25(11):1275-1286.
[15]Madamanchi NR,Vendrov A,Runge MS.Oxidative stress and vascular disease[J].Arterioscler Thromb Vasc Biol,2005,25(1):29-38.
[16]Bauersachs J,Fraccarollo D.Endothelial NO synthase target of aldosterone[J].Hypertension,2006,48(1):165-171.
[17]Galley JC,Straub AC.Redox Control of Vascular Function[J].Arterioscler Thromb Vasc Biol,2017,37(12):e178-e184.
[18]F?rstermann U,Xia N,Li H.Roles of Vascular Oxidative Stress and Nitric Oxide in the Pathogenesis of Atherosclerosis[J].Circ Res,2017,120(4):713-735.
[19]Chen L,Wu F,Xia WH,et al.CXCR4 gene transfer contributes to in vivo reendothelialization capacity of endothelial progenitor cells[J].Cardiovasc Res,2010,88(3):462-470.
[20]Wang JM,Yang Z,Xu MG,et al.Berberine-induced decline in circulating CD31+/CD42-microparticles is associated with improvement of endothelial function in humans[J].Eur J Pharmacol,2009,614(1-3):77-83.
[21]Milliez P,Girerd X,Plouin PF,et al.Evidence for an increased rate of cardiovascular events in patients with primary aldosteronism[J].J Am Coll Cardiol,2005,45(8):1243-1248.