CYP2J2及EETs通过抗氧化应激和抗凋亡减轻肺缺血再灌注损伤
|
摘要
目的探讨CYP2J2及其代谢产物EETs对肺缺血再灌注损伤的作用及其机制。方法将CYP2J2基因稳定转染至大鼠肺组织。分别构建大鼠肺缺血再灌注损伤模型和人肺动脉内皮细胞缺氧复氧模型,观察CYP2J2基因转染及其代谢产物EETs在体内和体外对氧化应激和凋亡相关指标的影响。结果在体内,CYP2J2基因过表达显著减轻了肺缺血再灌注损伤导致的氧化应激和凋亡。在体外,CYP2J2基因成功转染至人肺动脉内皮细胞,并增加CYP2J2蛋白表达。在体外缺氧复氧模型中,外源性EETs可以增强细胞活力,抑制细胞内活性氧的产生,减轻线粒体功能障碍,减轻多种凋亡事件。这些事件包括NADPH氧化酶的活化、线粒体跨膜电位衰竭、促凋亡蛋白和Caspase-3的活化。结论CYP2J2基因过表达及外源性EETs能够通过抗氧化应激和凋亡减轻肺缺血再灌注损伤,其作用可能是通过激活PI3K/Akt信号通路来介导的。
Objective To explore the effect of CYP2J2 and EETs on lung ischemia-reperfusion injury and the related mechanism.Methods CYP2J2 was stably transfected into lung tissue of rats.A rat model of lung ischemia-reperfusion injury and a human pulmonary artery endothelial cell(HPAEC)model of hypoxia-reoxygenation were established to observe the effect of CYP2J2 overexpression and EETs on oxidation stress and apoptosis induced by lung ischemia-reperfusion in vivo and in vitro.Results CYP2J2 overexpression significantly inhibited the level of oxidation stress and apoptosis in vivoinduced by lung ischemia-reperfusion.Moreover,CYP2J2 gene was successfully transfected into human pulmonary artery endothelial cells,leading to overexpression of CYP2J2 protein.In the HPAEC-based hypoxia-reoxygenation model,exogenous EETs enhanced cell viability,inhibited the production of intracellular reactive oxygen species,mitigated mitochondrial dysfunction and attenuated several apoptotic related events,including collapse of mitochondrial transmembrane potential,activation of NADPH oxidase,pro-apoptotic protein and Caspase-3.Conclusion CYP2J2 overexpression and exogenous EETs can protect against lung ischemiareperfusion injury via anti-oxidative stress and antiapoptosis in vivo and in vitro.These effects may be mediated by PI3 K/Akt signaling pathway.
Objective To explore the effect of CYP2J2 and EETs on lung ischemia-reperfusion injury and the related mechanism.Methods CYP2J2 was stably transfected into lung tissue of rats.A rat model of lung ischemia-reperfusion injury and a human pulmonary artery endothelial cell(HPAEC)model of hypoxia-reoxygenation were established to observe the effect of CYP2J2 overexpression and EETs on oxidation stress and apoptosis induced by lung ischemia-reperfusion in vivo and in vitro.Results CYP2J2 overexpression significantly inhibited the level of oxidation stress and apoptosis in vivoinduced by lung ischemia-reperfusion.Moreover,CYP2J2 gene was successfully transfected into human pulmonary artery endothelial cells,leading to overexpression of CYP2J2 protein.In the HPAEC-based hypoxia-reoxygenation model,exogenous EETs enhanced cell viability,inhibited the production of intracellular reactive oxygen species,mitigated mitochondrial dysfunction and attenuated several apoptotic related events,including collapse of mitochondrial transmembrane potential,activation of NADPH oxidase,pro-apoptotic protein and Caspase-3.Conclusion CYP2J2 overexpression and exogenous EETs can protect against lung ischemiareperfusion injury via anti-oxidative stress and antiapoptosis in vivo and in vitro.These effects may be mediated by PI3 K/Akt signaling pathway.
引文
[1]den Hengst W A,Gielis J F,Lin J Y,et al.Lung ischemiareperfusion injury:a molecular and clinical view on a complex pathophysiological process[J].Am J Physiol Heart Circ Physiol,2010,299(5):H1283-H1299.
[2]Wu S,Moomaw C R,Tomer K B,et al.Molecular cloning and expression of CYP2J2,a human cytochrome P450arachidonic acid epoxygenase highly expressed in heart[J].J Biol Chem,1996,271(7):3460-3468.
[3]Spector A A,Fang X,Snyder G D,et al.Epoxyeicosatrienoic acids(EETs):metabolism and biochemical function[J].Prog Lipid Res,2004,43(1):55-90.
[4]Li R,Xu X,Chen C,et al.Cytochrome P4502J2is protective against global cerebral ischemia in transgenic mice[J].Prostag Oth Lipid M,2012,99(3/4):68-78.
[5]Yang B,Graham L,Dikalov S,et al.Overexpression of cytochrome P450CYP2J2protects against hypoxia-reoxygenation injury in cultured bovine aortic endothelial cells[J].Mol Pharmacol,2001,60(2):310-320.
[6]Edin M L,Wang Z,Bradbury J A,et al.Endothelial expression of human cytochrome P450epoxygenase CYP2C8increases susceptibility to ischemia-reperfusion injury in isolated mouse heart[J].FASEB J,2011,25(10):3436-3447.
[7]杨世疆,王家宁,平伟,等.大鼠急性肺缺血再灌注损伤模型的建立与评估[J].中国医药导刊,2013,15(5):858-859,862.
[8]Chen W,Yang S,Ping W,et al.CYP2J2and EETs protect against lung ischemia/reperfusion injury via anti-inflammatory effects in vivo and in vitro[J].Cell Physiol Biochem,2015,35(5):2043-2054.
[9]Devasagayam T P,Tilak J C,Boloor K K,et al.Free radicals and antioxidants in human health:current status and future prospects[J].J Assoc Physicians India,2004,52:794-804.
[10]Kroemer G,Dallaporta B,Resche-Rigon M.The mitochondrial death/life regulator in apoptosis and necrosis[J].Annu Rev Physiol,1998,60(1):619-642.
[11]Esme H,Fidan H,Koken T,et al.Effect of lung ischemia--reperfusion on oxidative stress parameters of remote tissues[J].Eur J Cardiothorac Surg,2006,29(3):294-298.
[12]Ng C S,Wan S,Yim A P.Pulmonary ischaemia-reperfusion injury:role of apoptosis[J].Eur Respir J,2005,25(2):356-363.
[13]Valko M,Leibfritz D,Moncol J,et al.Free radicals and antioxidants in normal physiological functions and human disease[J].Int J Biochem Cell Biol,2007,39(1):44-84.
[14]Orrenius S.Reactive oxygen species in mitochondria-mediated cell death[J].Drug Metab Rev,2007,39(2/3):443-455.
[15]Frey R S,Ushio-Fukai M,Malik A B.NADPH oxidase-dependent signaling in endothelial cells:role in physiology and pathophysiology[J].Antioxid Redox Sign,2009,11(4):791-810.
[16]Zhu C,Bilali A,Georgieva G S,et al.Salvage of nonischemic control lung from injury by unilateral ischemic lung with apocynin,a nicotinamide adenine dinucleotide phosphate(NADPH)oxidase inhibitor,in isolated perfused rat lung[J].Transl Res,2008,152(6):273-282.
[17]Dodd-o J M,Welsh L E,Salazar J D,et al.Effect of NADPH oxidase inhibition on cardiopulmonary bypass-induced lung injury[J].Am J Physiol Heart Circ Physiol,2004,287(2):H927-H936.
[18]Feng W,Xu X,Zhao G,et al.EETs and CYP2J2inhibit TNFalpha-induced apoptosis in pulmonary artery endothelial cells and TGF-beta1-induced migration in pulmonary artery smooth muscle cells[J].Int J Mol Med,2013,32(3):685-693.
[19]Zhou J,Zhang S,Zhao X,et al.Melatonin impairs NADPH oxidase assembly and decreases superoxide anion production in microglia exposed to amyloid-beta1-42[J].J Pineal Res,2008,45(2):157-165.
[20]Yang S,Lin L,Chen J X,et al.Cytochrome P-450epoxygenases protect endothelial cells from apoptosis induced by tumor necrosis factor-alpha via MAPK and PI3K/Akt signaling pathways[J].Am J Physiol Heart Circ Physiol,2007,293(1):H142-H151.
[2]Wu S,Moomaw C R,Tomer K B,et al.Molecular cloning and expression of CYP2J2,a human cytochrome P450arachidonic acid epoxygenase highly expressed in heart[J].J Biol Chem,1996,271(7):3460-3468.
[3]Spector A A,Fang X,Snyder G D,et al.Epoxyeicosatrienoic acids(EETs):metabolism and biochemical function[J].Prog Lipid Res,2004,43(1):55-90.
[4]Li R,Xu X,Chen C,et al.Cytochrome P4502J2is protective against global cerebral ischemia in transgenic mice[J].Prostag Oth Lipid M,2012,99(3/4):68-78.
[5]Yang B,Graham L,Dikalov S,et al.Overexpression of cytochrome P450CYP2J2protects against hypoxia-reoxygenation injury in cultured bovine aortic endothelial cells[J].Mol Pharmacol,2001,60(2):310-320.
[6]Edin M L,Wang Z,Bradbury J A,et al.Endothelial expression of human cytochrome P450epoxygenase CYP2C8increases susceptibility to ischemia-reperfusion injury in isolated mouse heart[J].FASEB J,2011,25(10):3436-3447.
[7]杨世疆,王家宁,平伟,等.大鼠急性肺缺血再灌注损伤模型的建立与评估[J].中国医药导刊,2013,15(5):858-859,862.
[8]Chen W,Yang S,Ping W,et al.CYP2J2and EETs protect against lung ischemia/reperfusion injury via anti-inflammatory effects in vivo and in vitro[J].Cell Physiol Biochem,2015,35(5):2043-2054.
[9]Devasagayam T P,Tilak J C,Boloor K K,et al.Free radicals and antioxidants in human health:current status and future prospects[J].J Assoc Physicians India,2004,52:794-804.
[10]Kroemer G,Dallaporta B,Resche-Rigon M.The mitochondrial death/life regulator in apoptosis and necrosis[J].Annu Rev Physiol,1998,60(1):619-642.
[11]Esme H,Fidan H,Koken T,et al.Effect of lung ischemia--reperfusion on oxidative stress parameters of remote tissues[J].Eur J Cardiothorac Surg,2006,29(3):294-298.
[12]Ng C S,Wan S,Yim A P.Pulmonary ischaemia-reperfusion injury:role of apoptosis[J].Eur Respir J,2005,25(2):356-363.
[13]Valko M,Leibfritz D,Moncol J,et al.Free radicals and antioxidants in normal physiological functions and human disease[J].Int J Biochem Cell Biol,2007,39(1):44-84.
[14]Orrenius S.Reactive oxygen species in mitochondria-mediated cell death[J].Drug Metab Rev,2007,39(2/3):443-455.
[15]Frey R S,Ushio-Fukai M,Malik A B.NADPH oxidase-dependent signaling in endothelial cells:role in physiology and pathophysiology[J].Antioxid Redox Sign,2009,11(4):791-810.
[16]Zhu C,Bilali A,Georgieva G S,et al.Salvage of nonischemic control lung from injury by unilateral ischemic lung with apocynin,a nicotinamide adenine dinucleotide phosphate(NADPH)oxidase inhibitor,in isolated perfused rat lung[J].Transl Res,2008,152(6):273-282.
[17]Dodd-o J M,Welsh L E,Salazar J D,et al.Effect of NADPH oxidase inhibition on cardiopulmonary bypass-induced lung injury[J].Am J Physiol Heart Circ Physiol,2004,287(2):H927-H936.
[18]Feng W,Xu X,Zhao G,et al.EETs and CYP2J2inhibit TNFalpha-induced apoptosis in pulmonary artery endothelial cells and TGF-beta1-induced migration in pulmonary artery smooth muscle cells[J].Int J Mol Med,2013,32(3):685-693.
[19]Zhou J,Zhang S,Zhao X,et al.Melatonin impairs NADPH oxidase assembly and decreases superoxide anion production in microglia exposed to amyloid-beta1-42[J].J Pineal Res,2008,45(2):157-165.
[20]Yang S,Lin L,Chen J X,et al.Cytochrome P-450epoxygenases protect endothelial cells from apoptosis induced by tumor necrosis factor-alpha via MAPK and PI3K/Akt signaling pathways[J].Am J Physiol Heart Circ Physiol,2007,293(1):H142-H151.