氧化苦参碱对人脐静脉内皮细胞体外缺血再灌注损伤的保护作用及其分子机制
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摘要
目的研究氧化苦参碱(oxymatrine,OMT)对人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVEC)体外缺血再灌注(ischemia reperfusion,I/R)损伤的保护作用及其分子机制。方法将HUVEC分为对照组、I/R组和OMT 0. 5、1、2μg/ml组,对照组为正常培养细胞,I/R组和OMT 0. 5、1、2μg/ml组均制备I/R损伤模型,OMT0. 5、1、2μg/ml组分别给予OMT 0. 5、1、2μg/ml处理。检测5组细胞存活率、乳酸脱氢酶(LDH)含量、细胞凋亡情况及Bcl-2、Bax、Caspase-3蛋白表达水平。结果与I/R组比较,OMT 0. 5、1、2μg/ml组HUVEC存活率升高,培养液中LDH含量降低,细胞凋亡减少,Bcl-2/Bax逐渐上调,Caspase-3蛋白表达下调,且呈剂量依赖性(P <0. 05,P <0. 01)。结论 OMT对HUVEC体外I/R损伤有保护作用,其机制可能与减少细胞凋亡有关。
Objective To study protective effect of Oxymatrine( OMT) on human umbilical vein endothelial cells( HUVEC) ischemia-reperfusion( I/R) injury in vitro and its molecular mechanism. Methods HUVEC were divided into control group,I/R group,0. 5,1 and 2μg/ml OMT groups. Control group was normal cultured cells. I/R group,0. 5,1 and 2μg/ml OMT groups were prepared I/R injury models. The 0. 5,1 and 2μg/ml OMT groups were treated with 0. 5,1 and 2μg/ml OMT respectively. Cell survival rate,lactate dehydrogenase( LDH) contents,conditions of cell apoptosis and expressions of Bcl-2,Bax and Caspase-3 proteins were detected among 5 groups. Results Compared with those in I/R group,in 0. 5,1 and 2μg/ml OMT groups,survival rates of HUVEC were increased; LDH contents in culture medium were decreased; numbers of cell apoptosis were decreased; values of Bcl-2/Bax were increased gradually; expressions of Caspase-3 proteins were decreased in a dose-dependent manner( P < 0. 05,P < 0. 01). Conclusion Oxymatrine has protective effects on HUVEC ischemia-reperfusion injury in vitro,and its mechanism may be related to reduction of apoptosis.
Objective To study protective effect of Oxymatrine( OMT) on human umbilical vein endothelial cells( HUVEC) ischemia-reperfusion( I/R) injury in vitro and its molecular mechanism. Methods HUVEC were divided into control group,I/R group,0. 5,1 and 2μg/ml OMT groups. Control group was normal cultured cells. I/R group,0. 5,1 and 2μg/ml OMT groups were prepared I/R injury models. The 0. 5,1 and 2μg/ml OMT groups were treated with 0. 5,1 and 2μg/ml OMT respectively. Cell survival rate,lactate dehydrogenase( LDH) contents,conditions of cell apoptosis and expressions of Bcl-2,Bax and Caspase-3 proteins were detected among 5 groups. Results Compared with those in I/R group,in 0. 5,1 and 2μg/ml OMT groups,survival rates of HUVEC were increased; LDH contents in culture medium were decreased; numbers of cell apoptosis were decreased; values of Bcl-2/Bax were increased gradually; expressions of Caspase-3 proteins were decreased in a dose-dependent manner( P < 0. 05,P < 0. 01). Conclusion Oxymatrine has protective effects on HUVEC ischemia-reperfusion injury in vitro,and its mechanism may be related to reduction of apoptosis.
引文
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[19]Zhang M,Wang X,Hou X,et al.Oxymatrineprotects against myocardial injury via inhibition of JAK2/STAT3signaling in rat septic shock[J].Mol Med Rep,2013,7:1293-1299.
[20]Jiang G,Liu X,Wang M,et al.Oxymatrine ameliorates renal ischemia-reperfusion injury from oxidative stress through Nrf2/HO-1 pathway[J].Acta Cir Bras,2015,30(6):422-429.
[21]程钢,秦媛媛,程迪.氧化苦参碱对大鼠局灶性脑缺血损伤的保护作用及其抑制凋亡的作用机制[J].中国药理学通报,2013,29(3):387-392.
[22]Zhu B,Yang J,Chen S,et al.Oxymatrine on Hsp90a expression and apoptosis in a model of lung ischemia-reperfusion injury[J].Exp Ther Med,2017,13(4):1381-1385.
[23]王雪芬,王磊,陈树杰.苦参素对大鼠心肌缺血再灌注损伤的保护作用及其机制研究[J].中国中医急症,2016,25(6):988-992.
[24]周隆书,赵国龙,李明亮,等.氧化苦参碱预处理对大鼠缺血再灌注损伤心肌保护作用及其机制研究[J].宁夏医科大学学报,2015,37(6):650-656.
[2]Ozturk H,Cetinkaya A,Yilmaz F,et al.Protective effect of oxymatrine against renal ischemia/reperfusion injury in rats[J].Bratisl Lek Listy,2017,118(4):217-222.
[3]Zhang X,Jiang W,Zhou A L,et al.Inhibitory effect of oxymatrine on hepatocyte apoptosis VIA TLR4/PI3K/Akt/GSK-3βsignaling pathway[J].World J Gastroenterol,2017,23(21):3839-3849.
[4]陈树杰.苦参素对缺氧/复氧损伤乳鼠心肌细胞保护作用研究[J].辽宁中医药大学学报,2016,18(9):49-53.
[5]Zhao J,Yu S,Tong L,et al.Oxymatrine attenuates intestinal ischemia/reperfusion injury in rats[J].Surg Today,2008,38(10):931-937.
[6]Zhu B,Yang J R,Chen S F,et al.The attenuation of lung ischemia reperfusion injury by oxymatrine[J].Cell Biochem Biophys,2014,70(1):333-336.
[7]Li S,Liu C,Gu L,et al.Autophagy protects cardiomyocytes from the myocardial ischaemia-reperfusion injury through the clearance of CLP36[J].Open Biology,2016,6(8).
[8]Scarabelli T,Stephanou A,Rayment N,et al.Apoptosis of endothelial cells precedes myocyte cell apoptosis in ischemia/reperfusion injury[J].Circulation,2001,104(3):253-256.
[9]牛丽丽,祝善俊,李鲁光.心肌缺血再灌注时内皮细胞损伤功能改变与心肌梗塞面积的关系[J].中华内科杂志,1998,37(11):16-18.
[10]Kumar P,Shen Q,Pivetti C D,et al.Molecular mechanisms of endothelial hyperpermeability:implications in inflammation[J].Expert Rev Mol Med,2009,11:e19.
[11]Zhang Y Q,Ding N,Zeng Y F,et al.New progress in roles of nitric oxide during hepatic ischemia reperfusion injury[J].World J Gastroenterol,2017,23(14):2505-2510.
[12]Guddeti R R,Prasad A,Matsuzawa Y,et al.Role of endothelin in microvascular dysfunction following percutaneous coronary intervention for non-ST elevation acute coronary syndromes:a single-centrerandomised controlled trial[J].Open Heart,2016,3(2):e000428.
[13]Barrabés J A,Inserte J,Mirabet M,et al.Antagonism of P2Y12 or GPIIb/IIIa receptors reduces platelet-mediated myocardial injury after ischaemia and reperfusion in isolated rat hearts[J].Thromb Haemost,2010,104(1):128-135.
[14]Nernpermpisooth N,Prompunt E,Kumphune S.An in vitro endothelial cell protective effect of secretory leukocyte protease inhibitor against simulated ischaemia/reperfusion injury[J].Exp Ther Med,2017,14(6):5793-5800.
[15]Liao L X,Zhao M B,Dong X,et al.TDB protects vascular endothelial cells against oxygen-glucose deprivation/reperfusion-induced injury by targeting miR-34a to increase Bcl-2 expression[J].Sci Rep,2016,6:37959.
[16]Kristen A V,Ackermann K,Buss S,et al.Inhibition of apoptosis by the intrinsic but not the extrinsic apoptotic pathway in myocardial ischemia-reperfusion[J].Cardiovasc Pathol,2013,22(4):280-286.
[17]范宗静,吴旸,唐杰.缺血再灌注损伤人心脏微血管内皮细胞凋亡基因Bcl-2、Bax的表达及黄芪多糖干预研究[J].中华中医药杂志,2017,32(12):5603-5606.
[18]Xiao T T,Wang Y Y,Zhang Y,et al.Similar to spironolactone,oxymatrine is protective in aldosterone-induced cardiomyocyte injury VIA inhibition of calpain and apoptosis-inducing factor signaling[J].PLo S One,2014,9(2):e88856.
[19]Zhang M,Wang X,Hou X,et al.Oxymatrineprotects against myocardial injury via inhibition of JAK2/STAT3signaling in rat septic shock[J].Mol Med Rep,2013,7:1293-1299.
[20]Jiang G,Liu X,Wang M,et al.Oxymatrine ameliorates renal ischemia-reperfusion injury from oxidative stress through Nrf2/HO-1 pathway[J].Acta Cir Bras,2015,30(6):422-429.
[21]程钢,秦媛媛,程迪.氧化苦参碱对大鼠局灶性脑缺血损伤的保护作用及其抑制凋亡的作用机制[J].中国药理学通报,2013,29(3):387-392.
[22]Zhu B,Yang J,Chen S,et al.Oxymatrine on Hsp90a expression and apoptosis in a model of lung ischemia-reperfusion injury[J].Exp Ther Med,2017,13(4):1381-1385.
[23]王雪芬,王磊,陈树杰.苦参素对大鼠心肌缺血再灌注损伤的保护作用及其机制研究[J].中国中医急症,2016,25(6):988-992.
[24]周隆书,赵国龙,李明亮,等.氧化苦参碱预处理对大鼠缺血再灌注损伤心肌保护作用及其机制研究[J].宁夏医科大学学报,2015,37(6):650-656.