ERK/CT-1通路对氧化应激致H9C2细胞凋亡的影响
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摘要
目的观察在氧化应激情况下心肌营养因子-1(CT-1)的表达情况及其对心肌细胞凋亡的影响,探讨其相关机制。方法以过氧化氢刺激H9C2细胞建立氧化损伤模型,N-乙酰半胱氨酸或CT-1小干扰RNA(si RNA)与过氧化氢共同孵育以确定活性氧类(ROS)对CT-1表达的影响,以及CT-1在细胞凋亡中的作用。以丝裂原活化细胞外信号调节激酶(MEK/ERK)特异性抑制剂PD98059预处理H9C2细胞,以确定过氧化氢诱导CT-1表达的机制。结果过氧化氢可剂量依赖性地诱导CT-1的过表达,N-乙酰半胱氨酸可显著抑制过氧化氢诱导的CT-1过表达。过氧化氢诱导的H9C2细胞凋亡可被N-乙酰半胱氨酸阻断,但CT-1 siRNA可加重过氧化氢诱导的H9C2细胞凋亡。过氧化氢可剂量依赖性地促进ERK的活化,PD98059预处理明显抑制过氧化氢诱导的CT-1上调。结论过氧化氢能活化ERK/CT-1通路,从而缓解氧化应激引起的细胞凋亡。
Objective To examine the expression profile of cardiotrophin-1(CT-1) in oxidative stress challenged H9C2 cells and potential role of ERK/CT-1 signal pathway in this pathological procedure. Methods H9C2 cell model of oxidative stress was established by coincubation with hydrogen peroxide. N-acetylcysteine or CT-1 siRNA was incubated to determine the effect of reactive oxygen species(ROS) on CT-1 expression and apoptosis. Mitogen-activated extracellular signal-regulated kinase(MEK/ERK) specific inhibitor PD98059 was utilized to determine the probable mechanisms. Results Hydrogen peroxide induced a dose-dependent increasing of CT-1 expression, which was significantly attenuated by N-acetylcysteine. H9C2 cell apoptosis was inhibited by N-acetylcysteine treatment but was accelerated by CT-1 siRNA. Hydrogen peroxide dose promoted the activation of ERK signal pathway, while pretreatment with PD98059 significantly inhibited Hydrogen peroxide-induced upregulated expression of CT-1. Conclusions Hydrogen peroxide alleviates cell apoptosis probably through activation of ERK/CT-1 pathway.
Objective To examine the expression profile of cardiotrophin-1(CT-1) in oxidative stress challenged H9C2 cells and potential role of ERK/CT-1 signal pathway in this pathological procedure. Methods H9C2 cell model of oxidative stress was established by coincubation with hydrogen peroxide. N-acetylcysteine or CT-1 siRNA was incubated to determine the effect of reactive oxygen species(ROS) on CT-1 expression and apoptosis. Mitogen-activated extracellular signal-regulated kinase(MEK/ERK) specific inhibitor PD98059 was utilized to determine the probable mechanisms. Results Hydrogen peroxide induced a dose-dependent increasing of CT-1 expression, which was significantly attenuated by N-acetylcysteine. H9C2 cell apoptosis was inhibited by N-acetylcysteine treatment but was accelerated by CT-1 siRNA. Hydrogen peroxide dose promoted the activation of ERK signal pathway, while pretreatment with PD98059 significantly inhibited Hydrogen peroxide-induced upregulated expression of CT-1. Conclusions Hydrogen peroxide alleviates cell apoptosis probably through activation of ERK/CT-1 pathway.
引文
[1]ZHOU J,GAO J,ZHANG X,et al.MicroRNA-340-5p functions downstream of cardiotrophin-1 to regulate cardiac eccentric hypertrophy and heart failure via target gene dystrophin[J].Int Heart J,2015,56(4):454-458.
[2]ROBADOR P A,SAN J G,RODRIGUEZ C,et al.HIF-1-mediated up-regulation of cardiotrophin-1 is involved in the survival response of cardiomyocytes to hypoxia[J].Cardiovasc Res,2011,92(2):247-255.
[3]SHENG Z,KNOWLTON K,CHEN J,et al.Cardiotrophin 1(CT-1)inhibition of cardiac myocyte apoptosis via a mitogen-activated protein kinase-dependent pathway.Divergence from downstream CT-1 signals for myocardial cell hypertrophy[J].J Biol Chem,1997,272(9):5783-5791.
[4]STEPHANOU A,BRAR B,HEADS R,et al.Cardiotrophin-1induces heat shock protein accumulation in cultured cardiac cells and protects them from stressful stimuli[J].J Mol Cell Cardiol,1998,30(4):849-855.
[5]BRAR B K,STEPHANOU A,LIAO Z,et al.Cardiotrophin-1can protect cardiac myocytes from injury when added both prior to simulated ischaemia and at reoxygenation[J].Cardiovasc Res,2001,51(2):265-274.
[6]TALWAR S,SQUIRE I B,DOWNIE P F,et al.Plasma N terminal pro-brain natriuretic peptide and cardiotrophin 1 are raised in unstable angina[J].Heart,2000,84(4):421-424.
[7]TALWAR S,SQUIRE I B,O’BRIEN R J,et al.Plasma cardiotrophin-1 following acute myocardial infarction:relationship with left ventricular systolic dysfunction[J].Clin Sci(Lond),2002,102(1):9-14.
[8]SONG K,WANG S,HUANG B,et al.Plasma cardiotrophin-1levels are associated with hypertensive heart disease:a metaanalysis[J].J Clin Hypertens(Greenwich),2014,16(9):686-692.
[9]FARIAS J G,MOLINA V M,CARRASCO R A,et al.Antioxidant therapeutic strategies for cardiovascular conditions associated with oxidative stress[J].Nutrients,2017,9(9):E966.
[10]LEE K H,LEE S R,CHO H,et al.Cardioprotective effects of PKG activation by soluble GC activator,BAY 60-2770,in ischemia-reperfusion-injured rat hearts[J].PLoS One,2017,12(7):p e0180207.
[11]DONG Z L,WANG Y,LI T F,et al.p42/p44 mitogen-activated protein kinases inhibit atrial natriuretic peptide mRNAtranscription in gp130-mediated hypertrophic ventricular myocytes[J].Asian Pac J Trop Med,2014,7(3):216-220.
[12]廖新学,王艳丽,郭瑞鲜,等.ERK1/2介导H2O2预处理对抗氧化应激损伤的保护作用[J].中国药理学通报,2008,24(9):1151-1156.
[13]ZHAO H N,WANG Y,JIANG M N,et al.Relation of Cardiotrophin-1(CT-1)and cardiac transcription factor GATA4expression in rat’s cardiac myocytes hypertrophy and apoptosis[J].Pathol Res Pract,2009,205(9):615-625.
[2]ROBADOR P A,SAN J G,RODRIGUEZ C,et al.HIF-1-mediated up-regulation of cardiotrophin-1 is involved in the survival response of cardiomyocytes to hypoxia[J].Cardiovasc Res,2011,92(2):247-255.
[3]SHENG Z,KNOWLTON K,CHEN J,et al.Cardiotrophin 1(CT-1)inhibition of cardiac myocyte apoptosis via a mitogen-activated protein kinase-dependent pathway.Divergence from downstream CT-1 signals for myocardial cell hypertrophy[J].J Biol Chem,1997,272(9):5783-5791.
[4]STEPHANOU A,BRAR B,HEADS R,et al.Cardiotrophin-1induces heat shock protein accumulation in cultured cardiac cells and protects them from stressful stimuli[J].J Mol Cell Cardiol,1998,30(4):849-855.
[5]BRAR B K,STEPHANOU A,LIAO Z,et al.Cardiotrophin-1can protect cardiac myocytes from injury when added both prior to simulated ischaemia and at reoxygenation[J].Cardiovasc Res,2001,51(2):265-274.
[6]TALWAR S,SQUIRE I B,DOWNIE P F,et al.Plasma N terminal pro-brain natriuretic peptide and cardiotrophin 1 are raised in unstable angina[J].Heart,2000,84(4):421-424.
[7]TALWAR S,SQUIRE I B,O’BRIEN R J,et al.Plasma cardiotrophin-1 following acute myocardial infarction:relationship with left ventricular systolic dysfunction[J].Clin Sci(Lond),2002,102(1):9-14.
[8]SONG K,WANG S,HUANG B,et al.Plasma cardiotrophin-1levels are associated with hypertensive heart disease:a metaanalysis[J].J Clin Hypertens(Greenwich),2014,16(9):686-692.
[9]FARIAS J G,MOLINA V M,CARRASCO R A,et al.Antioxidant therapeutic strategies for cardiovascular conditions associated with oxidative stress[J].Nutrients,2017,9(9):E966.
[10]LEE K H,LEE S R,CHO H,et al.Cardioprotective effects of PKG activation by soluble GC activator,BAY 60-2770,in ischemia-reperfusion-injured rat hearts[J].PLoS One,2017,12(7):p e0180207.
[11]DONG Z L,WANG Y,LI T F,et al.p42/p44 mitogen-activated protein kinases inhibit atrial natriuretic peptide mRNAtranscription in gp130-mediated hypertrophic ventricular myocytes[J].Asian Pac J Trop Med,2014,7(3):216-220.
[12]廖新学,王艳丽,郭瑞鲜,等.ERK1/2介导H2O2预处理对抗氧化应激损伤的保护作用[J].中国药理学通报,2008,24(9):1151-1156.
[13]ZHAO H N,WANG Y,JIANG M N,et al.Relation of Cardiotrophin-1(CT-1)and cardiac transcription factor GATA4expression in rat’s cardiac myocytes hypertrophy and apoptosis[J].Pathol Res Pract,2009,205(9):615-625.