心肌肽对阿霉素诱导心肌细胞H9c2损伤的影响
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摘要
目的探讨心肌肽(cardiomyopeptide,Car)对阿霉素(doxorubicin,DOX)诱导的大鼠心肌细胞H9c2损伤的保护作用及其可能机制。方法 MTT法检测不同浓度(0、10、20、40 mg·L~(-1))心肌肽分别预处理6、8、12 h后,对DOX损伤心肌细胞存活率的影响; Western blot法检测心肌肽对DOX诱导心肌细胞中caspase-3、Bcl-2、Bax、IGF~(-1)R和IGFBP-3蛋白表达影响。结果随着心肌肽预处理时间增加,细胞存活率明显提高,且呈剂量依赖性;心肌肽40 mg·L~(-1)预处理8 h,使DOX的IC50值由(1. 2±0. 4)μmol·L~(-1)右移至(2. 3±0. 2)μmol·L~(-1); Western blot分析表明,经心肌肽处理后,Bax、caspase-3和IGFBP-3蛋白表达降低,Bcl-2和IGF~(-1)R蛋白表达增加,呈明显的剂量依赖性。结论心肌肽能保护心肌细胞,对抗DOX诱导的心肌细胞损伤,其机制可能与下调Bax和IGFBP-3,上调Bcl-2和IGF~(-1)R蛋白表达,抑制caspase-3活性有关。
Aim To investigate the effect of cardiomyopeptide on doxorubicin-induced toxicity on H9 c2 cardiomyoblasts and its related mechanism. Methods H9 c2 cells were respectively pretreated with different concentrations( 0,10,20,40 mg · L~(-1)) of cardiomyopeptide for 6 h,8 h,12 h. Cell viability was determined by MTT assay. The protein expression of caspase-3,Bcl-2,Bax,IGF~(-1) R and IGFBP-3 were detected by Western blot. Results Cardiomyopeptide protected H9 c2 cardiomyocytes from doxorubicin induced apoptosis in a dose-and time-dependent manner.The half maximal inhibitory concentration( IC50) wasshifted from( 1. 2 ± 0. 4) μmol · L~(-1) to( 2. 3 ± 0. 2)μmol·L~(-1). The expression of Bcl-2,IGF~(-1) R was upregulated,and Bax,IGFBP-3 and caspase-3 decreased in H9 c2 cells. Conclusions Cardiomyopeptide could prevent from apoptosis induced by doxorubicin in H9 c2 cells via increasing expression of IGF~(-1) R,thus up-regulation of the antiapoptotic protein Bcl-2 and inhibiting caspase-3 activity.
Aim To investigate the effect of cardiomyopeptide on doxorubicin-induced toxicity on H9 c2 cardiomyoblasts and its related mechanism. Methods H9 c2 cells were respectively pretreated with different concentrations( 0,10,20,40 mg · L~(-1)) of cardiomyopeptide for 6 h,8 h,12 h. Cell viability was determined by MTT assay. The protein expression of caspase-3,Bcl-2,Bax,IGF~(-1) R and IGFBP-3 were detected by Western blot. Results Cardiomyopeptide protected H9 c2 cardiomyocytes from doxorubicin induced apoptosis in a dose-and time-dependent manner.The half maximal inhibitory concentration( IC50) wasshifted from( 1. 2 ± 0. 4) μmol · L~(-1) to( 2. 3 ± 0. 2)μmol·L~(-1). The expression of Bcl-2,IGF~(-1) R was upregulated,and Bax,IGFBP-3 and caspase-3 decreased in H9 c2 cells. Conclusions Cardiomyopeptide could prevent from apoptosis induced by doxorubicin in H9 c2 cells via increasing expression of IGF~(-1) R,thus up-regulation of the antiapoptotic protein Bcl-2 and inhibiting caspase-3 activity.
引文
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[3]De Angelis A,Piegari E,Cappetta D,et al.Anthracycline cardiomyopathy is mediated by depletion of the cardiac stem cell pool and is rescued by restoration of progenitor cell function[J].Circulation,2010,121(2):276-92.
[4]Piegari E,De Angelis A,Cappetta D,et al.Doxorubicin induces senescence and impairs function of human cardiac progenitor cells[J].Basic Res Cardiol,2013,108(2):334.
[5]Esaki M,Takemura G,Kosai K,et al.Treatment with an adenoviral vector encoding hepatocyte growth factor mitigates established cardiac dysfunction in doxorubicin-induced cardiomyopathy[J].Am J Physiol Heart Circ Physiol,2008,294(2):1048-57.
[6]杨立平,陈良万,张贵灿,等.注射用心肌肽预处理对大鼠未成熟心肌缺血/再灌注损伤的保护作用及其机制[J].吉林大学学报(医学版),2011,37(6):1037-42.[6]Yang L P,Chen L W,Zhang G C,et al.Protective effect of cardiomypeotidin for injection preconditioning on immature myocardial ischemia-reperfusion injury in young rats and its mechanism[J].JJilin Univ(Med Ed),2011,37(6):1037-42.
[7]阮鹏,潘志锋,舟玲善,等.心肌肽素在心脏手术中对心肌的保护作用[J].中华胸心血管外科杂志,2001,17(5):288-90.[7]Ruan P,Pan Z F,Zhou L S,et al.Protective effect of cardiomyopeptidin on myocardial injuries during open heart operation[J].Chin J Thorac Cardiovasc Surg,2001,17(5):288-90.
[8]周宇,李晶,鲍翠玉.姜黄素纳米粒对高脂诱导的心肌细胞损伤的作用[J].中国药理学通报,2018,34(9):1283-8.[8]Zhou Y,Li J,Bao C Y.Effects of curcumin nanoparticles on hyperlipid-induced cardiomyocyte injury[J].Chin Pharmacol Bull,2018,34(9):1283-8.
[9]张军,谷翔,黄问银,等.GLP-1对AGEs诱导H9C2心肌细胞凋亡的保护作用研究[J].中国药理学通报,2017,33(1):120-6[9]Zhang J,Gu X,Huang W Y,et al.Protective effect of GLP-1 against AGEs-induced H9C2 myocardial cell apoptosis[J].Chin Pharmacol Bull,2017,33(1):120-6.
[10]董雅洁,高维娟.bcl-2、Bax、caspase-3在细胞凋亡中的作用及其关系[J].中国老年学杂志,2012,32(21):4828-30.[10]Dong Y J,Gao W J.Reciprocal effect of bcl-2,bax and caspase-3on apoptosis[J].Chin J Gerontol,2012,32(21):4828-30.
[11]Wang L,Ma W,Markovich R,et al.Regulation of cardiomyocyte apoptotic signaling by insulin-like growth factor I[J].Circ Res,1998,83(5):516-22.
[12]Morales M P,Galvez A,Eltit J M,et al.IGF-1 regulates apoptosis of cardiac myocyte induced by osmotic-stress[J].Biochem Biophys Res Commun,2000,270(3):1029-35.
[13]Wang L,Ma W,Markovich R,et al.Insulin-like growth factor Imodulates induction of apoptotic signaling in H9C2 cardiac muscle cells[J].Endocrinology,1998,139(3):1354-60.
[14]Jogie-Brahim S,Feldman D,Oh Y.Unraveling insulin-like growth factor binding protein-3 actions in human disease[J].Endocr Rev,2009,30(5):417-37.
[15]Patrizia F,Paolo S,Silvano G,et al.Doxorubicin impairs the insulin-like growth factor-1 system and causes insulin-like growth factor-1 resistance in cardiomyocytes[J].PLoS One,2015,10(5):e0124643.