银杏内酯B保护缺氧/复氧诱导心肌细胞损伤模型的作用及机制
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摘要
背景:银杏内酯B是银杏叶提取物,被认为具有抑制细胞凋亡、抗血小板聚集等多种作用,被广泛用于治疗心脑血管疾病,但其具体作用机制不明。目的:探讨银杏内酯B对缺氧/复氧诱导心肌细胞损伤的保护作用及其可能机制。方法:分别用普通培养基和含一定浓度银杏内酯B的培养基培养H9C2心肌细胞(各2份),再分别向其中1份预先加入ERK抑制剂处理后再经过缺氧/复氧(H/R)干预,并以此分为对照组、银杏组、对照+ERK抑制组及银杏+ERK抑制组,检测各组的凋亡率、增殖率及p-ERK1/2蛋白表达。结果与结论:①与对照组相比,银杏组的细胞凋亡率降低、增殖率增高,p-ERK1/2蛋白表达增加(P <0.05),②与银杏组相比,银杏+ERK抑制组的细胞凋亡率增加,增殖率降低,p-ERK1/2蛋白表达显著降低(P<0.05);③与对照+ERK抑制组相比,银杏+ERK抑制组的细胞凋亡率有所降低、增殖率增加(P <0.05),但p-ERK1/2蛋白表达无统计学差异;④结果说明,银杏内酯B能够降低缺氧/复氧引起的心肌细胞凋亡,提高存活,进而保护心肌,其机制可能通过银杏内酯增加心肌细胞中p-ERK1/2蛋白表达有关。
BACKGROUND: Ginkgolide B is extracted form ginkgo, which has been shown to inhibit cell apoptosis and platelet aggregation and has been extensively applied in the treatment of cardiovascular and cerebrovascular diseases. However, the underlying mechanism remains unclear. OBJECTIVE: To observe the protective effect of ginkgolide B on hypoxia/reoxygenation-induced cardiomyocyte injury and its underlying mechanisms. METHODS: H9C2 myocardial cells were divided into control group(common medium), ginkgolide B group(medium containing ginkgolide B), control plus ERK inhibitor group(common medium, ERK inhibitor prior to hypoxia/reoxygenation) and ginkgolide B plus ERK inhibitor group(medium containing ginkgolide B, ERK inhibitor prior to hypoxia/reoxygenation). The apoptosis rate, proliferation rate and p-ERK1/2 protein expression were detected. RESULTS AND CONCLUSION:(1) Compared with the control group, the apoptosis rate was significantly decreased, and the proliferation rate and the p-ERK1/2 protein expression were significantly increased in the ginkgolide B group(P < 0.05).(2) Compared with the ginkgolide B group, the apoptosis rate was significantly increased, and the proliferation rate and the p-ERK1/2 protein expression were significantly decreased in the ginkgolide B plus ERK inhibitor group(P < 0.05).(3) Compared with the control plus ERK inhibitor group, the apoptosis rate was significantly decreased and the proliferation rate was significantly increased in the ginkgolide B plus ERK inhibitor group(P < 0.05), and the p-ERK1/2 protein expression showed no significant difference.(4) These results indicate that ginkgolide B can reduce the cell apoptosis of hypoxia/reoxygenation-induced cardiomyocyte injury, and improve cell survival, thereby protecting the myocardium, which is by increasing p-ERK1/2 protein expression.
BACKGROUND: Ginkgolide B is extracted form ginkgo, which has been shown to inhibit cell apoptosis and platelet aggregation and has been extensively applied in the treatment of cardiovascular and cerebrovascular diseases. However, the underlying mechanism remains unclear. OBJECTIVE: To observe the protective effect of ginkgolide B on hypoxia/reoxygenation-induced cardiomyocyte injury and its underlying mechanisms. METHODS: H9C2 myocardial cells were divided into control group(common medium), ginkgolide B group(medium containing ginkgolide B), control plus ERK inhibitor group(common medium, ERK inhibitor prior to hypoxia/reoxygenation) and ginkgolide B plus ERK inhibitor group(medium containing ginkgolide B, ERK inhibitor prior to hypoxia/reoxygenation). The apoptosis rate, proliferation rate and p-ERK1/2 protein expression were detected. RESULTS AND CONCLUSION:(1) Compared with the control group, the apoptosis rate was significantly decreased, and the proliferation rate and the p-ERK1/2 protein expression were significantly increased in the ginkgolide B group(P < 0.05).(2) Compared with the ginkgolide B group, the apoptosis rate was significantly increased, and the proliferation rate and the p-ERK1/2 protein expression were significantly decreased in the ginkgolide B plus ERK inhibitor group(P < 0.05).(3) Compared with the control plus ERK inhibitor group, the apoptosis rate was significantly decreased and the proliferation rate was significantly increased in the ginkgolide B plus ERK inhibitor group(P < 0.05), and the p-ERK1/2 protein expression showed no significant difference.(4) These results indicate that ginkgolide B can reduce the cell apoptosis of hypoxia/reoxygenation-induced cardiomyocyte injury, and improve cell survival, thereby protecting the myocardium, which is by increasing p-ERK1/2 protein expression.
引文
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[7]Li MY,Chang CT,Han YT,et al.Ginkgolide B promotes neuronal differentiation through the Wnt/β-catenin pathway in neural stem cells of the postnatal mammalian subventricular zone.Sci Rep2018;8(1):14947.
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[14] Xu Z,Sun J,Tong Q,et al.The Role of ERK1/2 in the Development of Diabetic Cardiomyopathy. Int J Mol Sci.2016;17(12).
[15] Liu P,Zhong TP.MAPK/ERK signalling is required for zebrafish cardiac regeneration.Biotechnol Lett.2017;39(7):1069-1077.
[16] Filippone SM,Samidurai A,Roh SK,et al.Reperfusion Therapy with Rapamycin Attenuates Myocardial Infarction through Activation of AKT and ERK.Oxid Med Cell Longev.2017;2017:4619720.
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[18] Duan J, Yang Y, Liu H, et al. Osthole ameliorates acute myocardial infarction in rats by decreasing the expression of inflammatoryrelated cytokines, diminishing MMP-2 expression and activating p-ERK.Int J Mol Med.2016;37(1):207-216.
[19] Liu Z,Chen JM,Huang H,et al.The protective effect of trimetazidine on myocardial ischemia/reperfusion injury through activating AMPK and ERK signaling pathway.Metabolism, 2016;65(3):122-130.
[20] Thomas CJ,Lim NR,Kedikaetswe A,et al.Evidence that the MEK/ERK but not the PI3K/Akt pathway is required for protection from myocardial ischemia-reperfusion injury by3',4'-dihydroxyflavonol. Eur J Pharmacol.2015;758:53-59.
[21] Zheng PD,Mungur R,Zhou HJ,et al.Ginkgolide B promotes the proliferation and differentiation of neural stem cells following cerebral ischemia/reperfusion injury, both in vivo and in vitro. Neural Regen Res.2018;13(7):1204-1211.
[22]杨鹏飞,陈卫东.银杏内酯B药理作用研究进展[J].安徽中医学院学报, 2012,31(5):86-90.
[23] Fang W,Deng Y,Li Y,et al.Blood brain barrier permeability and therapeutic time window of Ginkgolide B in ischemia-reperfusion injury.Eur J Pharm Sci.2010;39(1-3):8-14.
[24] Gu JH,Ge JB,Li M,et al.Inhibition of NF-κB activation is associated with anti-inflammatory and anti-apoptotic effects of Ginkgolide B in a mouse model of cerebral ischemia/reperfusion injury. Eur J Pharm Sci.2012;47(4):652-660.
[25] Shu ZM, Shu XD, Li HQ, et al. Ginkgolide B Protects Against Ischemic Stroke Via Modulating Microglia Polarization in Mice. CNS Neurosci Ther 2016;22(9):729-739.
[26] Hao Y,Sun Y,Xu C,et al.Improvement of contractile function in isolated cardiomyocytes from ischemia-reperfusion rats by ginkgolide B pretreatment. J Cardiovasc Pharmacol. 2009;54(1):3-9.
[27]姜秀新,管斯斯,鲍利,等.银杏内酯B对胶原或二磷酸腺苷刺激血小板活化后一氧化氮含量及血管舒张因子刺激磷酸蛋白磷酸化的影响[J].中国组织工程研究与临床康复, 2009,13(11):2066-2070.
[2]González-Montero J,Brito R, Gajardo AI, et al. Myocardial reperfusion injury and oxidative stress:Therapeutic opportunities.World J Cardiol.2018;10(9):74-86.
[3]Frangogiannis NG. Pathophysiology of Myocardial Infarction.Compr Physiol.2015;5(4):1841-1875.
[4]Guo Y, Luo F, Liu Q, et al. Regulatory non-coding RNAs in acute myocardial infarction.J Cell Mol Med.2017;21(5):1013-1023.
[5]Roberts RE.The extracellular signal-regulated kinase(ERK)pathway:a potential therapeutic target in hypertension.J Exp Pharmacol.2012;4:77-83.
[6]Pei HX,Hua R,Guan CX,et al.Ginkgolide B Reduces the Degradation of Membrane Phospholipids to Prevent Ischemia/Reperfusion Myocardial Injury in Rats.Pharmacology. 2015;96(5-6):233-239.
[7]Li MY,Chang CT,Han YT,et al.Ginkgolide B promotes neuronal differentiation through the Wnt/β-catenin pathway in neural stem cells of the postnatal mammalian subventricular zone.Sci Rep2018;8(1):14947.
[8]Januzzi JL,Filippatos G,Nieminen M,et al.Troponin elevation in patients with heart failure:on behalf of the third Universal Definition of Myocardial Infarction Global Task Force:Heart Failure Section.Eur Heart J.2012;33(18):2265-2271.
[9]Martin LM,Januzzi JL,Thompson RW,et al.Clinical Profile of Acute Myocardial Infarction Patients Included in the Hospital Readmissions Reduction Program.J Am Heart Assoc.2018;7(16):e009339.
[10] Durak I,Kudaiberdieva G,Gorenek B.Prognostic implications of arrhythmias during primary percutaneous coronary interventions for ST-elevation myocardial infraction.Expert Rev Cardiovasc Ther.2015;13(1):85-94.
[11] Pan J, Alimujiang M, Chen Q, et al. Exosomes derived from miR-146a-modified adipose-derived stem cells attenuate acute myocardial infarction-induced myocardial damage via downregulation of early growth response factor 1.J Cell Biochem.2018;
[12] Wang A,Zhang H,Liang Z,et al.U0126 attenuates ischemia/reperfusion-induced apoptosis and autophagy in myocardium through MEK/ERK/EGR-1 pathway.Eur J Pharmacol.2016;788:280-285.
[13] Traister A,Li M,Aafaqi S,et al.Integrin-linked kinase mediates force transduction in cardiomyocytes by modulating SERCA2a/PLN function.Nat Commun.2014;5:4533.
[14] Xu Z,Sun J,Tong Q,et al.The Role of ERK1/2 in the Development of Diabetic Cardiomyopathy. Int J Mol Sci.2016;17(12).
[15] Liu P,Zhong TP.MAPK/ERK signalling is required for zebrafish cardiac regeneration.Biotechnol Lett.2017;39(7):1069-1077.
[16] Filippone SM,Samidurai A,Roh SK,et al.Reperfusion Therapy with Rapamycin Attenuates Myocardial Infarction through Activation of AKT and ERK.Oxid Med Cell Longev.2017;2017:4619720.
[17] Hausenloy DJ,Yellon DM.New directions for protecting the heart against ischaemia-reperfusion injury:targeting the Reperfusion Injury Salvage Kinase(RISK)-pathway.Cardiovasc Res.2004;61(3):448-460.
[18] Duan J, Yang Y, Liu H, et al. Osthole ameliorates acute myocardial infarction in rats by decreasing the expression of inflammatoryrelated cytokines, diminishing MMP-2 expression and activating p-ERK.Int J Mol Med.2016;37(1):207-216.
[19] Liu Z,Chen JM,Huang H,et al.The protective effect of trimetazidine on myocardial ischemia/reperfusion injury through activating AMPK and ERK signaling pathway.Metabolism, 2016;65(3):122-130.
[20] Thomas CJ,Lim NR,Kedikaetswe A,et al.Evidence that the MEK/ERK but not the PI3K/Akt pathway is required for protection from myocardial ischemia-reperfusion injury by3',4'-dihydroxyflavonol. Eur J Pharmacol.2015;758:53-59.
[21] Zheng PD,Mungur R,Zhou HJ,et al.Ginkgolide B promotes the proliferation and differentiation of neural stem cells following cerebral ischemia/reperfusion injury, both in vivo and in vitro. Neural Regen Res.2018;13(7):1204-1211.
[22]杨鹏飞,陈卫东.银杏内酯B药理作用研究进展[J].安徽中医学院学报, 2012,31(5):86-90.
[23] Fang W,Deng Y,Li Y,et al.Blood brain barrier permeability and therapeutic time window of Ginkgolide B in ischemia-reperfusion injury.Eur J Pharm Sci.2010;39(1-3):8-14.
[24] Gu JH,Ge JB,Li M,et al.Inhibition of NF-κB activation is associated with anti-inflammatory and anti-apoptotic effects of Ginkgolide B in a mouse model of cerebral ischemia/reperfusion injury. Eur J Pharm Sci.2012;47(4):652-660.
[25] Shu ZM, Shu XD, Li HQ, et al. Ginkgolide B Protects Against Ischemic Stroke Via Modulating Microglia Polarization in Mice. CNS Neurosci Ther 2016;22(9):729-739.
[26] Hao Y,Sun Y,Xu C,et al.Improvement of contractile function in isolated cardiomyocytes from ischemia-reperfusion rats by ginkgolide B pretreatment. J Cardiovasc Pharmacol. 2009;54(1):3-9.
[27]姜秀新,管斯斯,鲍利,等.银杏内酯B对胶原或二磷酸腺苷刺激血小板活化后一氧化氮含量及血管舒张因子刺激磷酸蛋白磷酸化的影响[J].中国组织工程研究与临床康复, 2009,13(11):2066-2070.