缺氧复氧对原代心肌细胞Wnt/β-catenin信号通路表达及细胞凋亡的影响
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- 英文论文题名：Effects ofhypoxia/reoxygenationcardiomyocyte models on the expression of Wnt/β-catenin signaling pathway and the relationship between apoptosis of cardiomyocytes and the expression of the signaling pathway
- 论文作者：刘玉 ; 胡迎春 ; 谭晓秋 ; 陈睦虎 ; 陈唐葶 ; 刘英 ; 李绍兰 ; 钟武
- 英文论文作者：LIU Yu ; HU Ying-chun ; TAN Xiao-qiu ; CHEN Mu-hu ; CHEN Tang-ting ; LIU Ying ; LI Shao-lan ; ZHONG Wu ; Department of emergency ; the Affiliated Hospital of Southwest Medical University ; Institute of cardiovascular research ; Southwest Medical University
- 年：2017
- 作者机构：西南医科大学附属医院急诊医学部;西南医科大学心血管研究所;
- 论文关键词：原代心肌细胞 ; 缺氧复氧模型 ; wnt/β-catenin信号通路
- 英文论文关键词：Cardiomyocytes of neonatal rat ; Wnt/β-catenin signaling pathway ; Models of hypoxia/reoxygenation
- 会议召开时间：2017-08-07
- 会议录名称：中国解剖学会第六届全国解剖学技术学术会议论文汇编
- 语种：中文
- 分类号：R54
- 学会代码：ZGJP
- 会议名称：中国解剖学会第六届全国解剖学技术学术会议
- 会议地点：中国江西九江
- 主办单位：中国解剖学会科技开发和咨询工作委员会
- 学会名称：中国解剖学会
- 页数：7
- 文件大小：2695k
- 原文格式：D
- 会议级别：全国
摘要
目的探讨缺氧复氧对心肌细胞wnt信号通路的作用及该通路的激动与抑制对心肌细胞凋亡的影响。方法分离培养新生SD大鼠原代心肌细胞,建立原代心肌细胞缺氧复氧模型。随机分为control组、缺氧复氧组(H/R组)、缺氧复氧+agonist 1组(H/R+A组)、缺氧复氧+DKK-1组(H/R+D组)。应用RT-PCR技术检测wnt信号通路关键基因表达水平,流式细胞技术检测心肌细胞凋亡率。结果心肌细胞缺氧8h,复氧2 h,相对于control组,wnt3a、β-catenin、c-mycm RNA表达升高(P<0.05),H/R组低于H/R+A组(P<0.05),高于H/R+D组(P<0.05)。流式细胞检测,H/R组早期及晚期凋亡率均高于control组(P<0.05),低于H/R+A组(P<0.05),高于H/R+D(P<0.05)。结论缺氧复氧激活wnt/β-catenin信号通路,抑制该通路可减少心肌细胞凋亡。
Objective To investigate the expression of wnt3 a,β-catenin and c-myc in cardiomyocyte of hypoxia/reoxygenationon Wnt/β-catenin signaling pathway and the relationship between apoptosis of cardiomyocytes and the expression of the signaling pathway.Methods Thecardiomyocytes of neonatal SD rats were isolated and cultured.The cardiomyocyteswere randomly divided into control group,hypoxia/reoxygenation group(H/R),hypoxia/reoxygenation group + agonist(H/R + A group),hypoxia/reoxygenation group + DKK-1(group H/R + D).RT-PCR were used to detect the expression of key genes in Wnt signaling pathway,and the apoptosis rate was detected by flow cytometry.Results compared with control group,the expression of Wnt3 a,β-catenin,c-myc m RNA were increased(P<0.05),and H/R group was lower than that of H/R + A group(P<0.05),and also was higher than that of H/R + D group(P<0.05),Compared with H/R + A group(P<0.05),the apoptosis rate of H/R group was higher than that of group control(P<0.05),and also was higher than that of H/R + D(P<0.05).Conclusion wnt/β-catenin signaling pathway can be activated with hypoxia/reoxygenation in Cardiomyocytes of neonatal SD rats,and the apoptosis of cardiomyocytes will be increased.
Objective To investigate the expression of wnt3 a,β-catenin and c-myc in cardiomyocyte of hypoxia/reoxygenationon Wnt/β-catenin signaling pathway and the relationship between apoptosis of cardiomyocytes and the expression of the signaling pathway.Methods Thecardiomyocytes of neonatal SD rats were isolated and cultured.The cardiomyocyteswere randomly divided into control group,hypoxia/reoxygenation group(H/R),hypoxia/reoxygenation group + agonist(H/R + A group),hypoxia/reoxygenation group + DKK-1(group H/R + D).RT-PCR were used to detect the expression of key genes in Wnt signaling pathway,and the apoptosis rate was detected by flow cytometry.Results compared with control group,the expression of Wnt3 a,β-catenin,c-myc m RNA were increased(P<0.05),and H/R group was lower than that of H/R + A group(P<0.05),and also was higher than that of H/R + D group(P<0.05),Compared with H/R + A group(P<0.05),the apoptosis rate of H/R group was higher than that of group control(P<0.05),and also was higher than that of H/R + D(P<0.05).Conclusion wnt/β-catenin signaling pathway can be activated with hypoxia/reoxygenation in Cardiomyocytes of neonatal SD rats,and the apoptosis of cardiomyocytes will be increased.
引文
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[21]Koval A,Purvanov V,Egger-Adam D,et al.Yellow submarine of the Wnt/Frizzled signaling:Submerging from the G protein harbor to the targets[J].Biochemical Pharmacology,2011,82(10):1311-1319.
[22]Ozhan G,Weidinger G.Wnt/β-catenin signaling in heart regeneration[J].Cell Regeneration,2015,4(1).
[23]Klaus A,Muller M,Schulz H,et al.Wnt/-catenin and Bmp signals control distinct sets of transcription factors in cardiac progenitor cells[J].Proceedings of the National Academy of Sciences,2012,109(27):10921-10926.
[24]Palevski D,Levin Kotler L P,Kain D,et al.Loss of Macrophage Wnt Secretion Improves Remodeling and Function After Myocardial Infarction in Mice[J].Journal of the American Heart Association,2017,6(1):e4387.
[25]Liu J,Wu X,Mitchell B,et al.A Small-Molecule Agonist of the Wnt Signaling Pathway[J].Angewandte Chemie International Edition,2005,44(13):1987-1990.
[26]Kuncewitch M,Yang W,Jacob A,et al.Stimulation of Wnt/β-catenin signaling pathway with Wnt agonist reduces organ injury after hemorrhagic shock[J].Journal of Trauma and Acute Care Surgery,2015,78(4):793-800.
[27]Bastakoty D,Saraswati S,Joshi P,et al.Temporary,Systemic Inhibition of the WNT/beta-Catenin Pathway promotes Regenerative Cardiac Repair following Myocardial Infarct[J].Cell Stem Cells Regen Med,2016,2(2).
[28]Tao J,Abudoukelimu M,Ma Y,et al.Secreted frizzled related protein 1protects H9C2 cells from hypoxia/re-oxygenation injury by blocking the Wnt signaling pathway[J].Lipids in Health and Disease,2016,15(1).
[29]Ming M,Wang S,Wu W,et al.Activation of Wnt/β-Catenin Protein Signaling Induces Mitochondria-mediated Apoptosis in Hematopoietic Progenitor Cells[J].Journal of Biological Chemistry,2012,287(27):22683-22690.
[30]Zhang Z,Deb A,Zhang Z,et al.Secreted frizzled related protein 2protects cells from apoptosis by blocking the effect of canonical Wnt3a[J].Journal of Molecular and Cellular Cardiology,2009,46(3):370-377.
[31]Korkmaz-Ic?z S,Radovits T,SzabóG.Targeting phosphodiesterase 5as a therapeutic option against myocardial ischaemia/reperfusion injury and for treating heart failure[J].British Journal of Pharmacology,2017.Epub ahead of print
[32]Agrawal V,Gupta J K,Qureshi S S,et al.Role of cardiac renin angiotensin system in ischemia reperfusion injury and preconditioning of heart[J].Indian Heart Journal,2016,68(6):856-861.
[33]Lin J C,Kuo W W,Baskaran R,et al.Enhancement of beta-catenin in cardiomyocytes suppresses survival protein expression but promotes apoptosis and fibrosis[J].Cardiol J,2016.
[34]Kim K,Pang K M,Evans M,et al.Overexpression of beta-catenin induces apoptosis independent of its transactivation function with LEF-1 or the involvement of major G1 cell cycle regulators[J].Mol Biol Cell,2000,11(10):3509-3523.
[35]Prensner J R,Chen W,Han S,et al.The Long Non-Coding RNA PCAT-1 Promotes Prostate Cancer Cell Proliferation through c Myc[J].Neoplasia,2014,16(11):900-908.
[2]Neri M,Riezzo I,Pascale N,et al.Ischemia/Reperfusion Injury following Acute Myocardial Infarction:A Critical Issue for Clinicians and Forensic Pathologists[J].Mediators of Inflammation,2017,2017:1-14.
[3]Korkmaz-Ic?z S,Radovits T,SzabóG.Targeting phosphodiesterase 5as a therapeutic option against myocardial ischaemia/reperfusion injury and for treating heart failure[J].British Journal of Pharmacology,2017.Epub ahead of print
[4]姬婷婷,徐岩,张建华,等.新生SD大鼠心肌细胞的原代培养及荧光鉴定[J].安徽医科大学学报,2014(01):120-122.
[5]储倩雯,李伟秋,鲁诗史,等.利用低氧/厌氧工作站建立H9c2细胞缺氧复氧模型的探讨[J].中国生化药物杂志,2015(11):11-14.
[6]Sun J,Wei T,Bai S,et al.Calcium-sensing receptor-mediated mitogenactivated protein kinase pathway improves the status of transplanted mouse embryonic stem cells in rats with acute myocardial infarction[J].Molecular and Cellular Biochemistry,2017.Epub ahead of print
[7]Ahmad S,Panda B P,Kohli K,et al.Folic acid ameliorates celecoxibcardiotoxicity in a doxorubicin heart failure rat model[J].Pharm Biol,2017,55(1):1295-1303.
[8]Chang G,Zhang D,Liu J,et al.Exenatide protects against hypoxia/reoxygenation-induced apoptosis by improving mitochondrial function in H9c2 cells[J].Experimental Biology and Medicine,2014,239(4):414-422.
[9]Jiang Y,Chang G,Wang Y,et al.Geniposide Prevents Hypoxia/Reoxygenation-Induced Apoptosis in H9c2 Cells:Improvement of Mitochondrial Dysfunction and Activation of GLP-1R and the PI3K/AKT Signaling Pathway[J].Cellular Physiology and Biochemistry,2016,39(1):407-421.
[10]曾石秀,廖伟.原代心肌细胞培养[J].赣南医学院学报,2012,32(2):167-169.
[11]Zhong H,Xin H,Wu L,et al.Salidroside Attenuates Apoptosis in Ischemic Cardiomyocytes:a Mechanism Through a MitochondriaDependent Pathway[J].Journal of Pharmacological Sciences,2010,114(4):399-408.
[12]Ennis I L,Escudero E M,Console G M,et al.Regression of Isoproterenol-Induced Cardiac Hypertrophy by Na+/H+Exchanger Inhibition[J].Hypertension,2003,41(6):1324-1329.
[13]Yuan Y,Pan S,Yang S L,et al.Antioxidant and cardioprotective effects of Ilex cornuta on myocardial ischemia injury[J].Chin J Nat Med,2017,15(2):94-104.
[14]Park E,Kang D,Kang J C,et al.Cardioprotective effect of KR-33889,a novel PARP inhibitor,against oxidative stress-induced apoptosis in H9c2 cells and isolated rat hearts[J].Archives of Pharmacal Research,2017.Epub ahead of print
[15]孙晓阳,王雁玲.Wnt信号通路与哺乳动物生殖[J].生物化学与生物物理进展,2003,30(2):180~184.
[16]Zhang Y,Li X M,Zhang F K,et al.Activation of canonical Wnt signaling pathway promotes proliferation and self-renewal of rat hepatic oval cell line WB-F344 in vitro[J].World J Gastroenterol,2008,14(43):6673-6680.
[17]Shangguan L,Ning G,Luo Z,et al.Fibulin-4 reduces extracellular matrix production and suppresses chondrocyte differentiation via DKK1-mediated canonical Wnt/beta-catenin signaling[J].Int J Biol Macromol,2017,99:293-299.
[18]Rashid S T,Humphries J D,Byron A,et al.Proteomic analysis of extracellular matrix from the hepatic stellate cell line LX-2 identifies CYR61 and Wnt-5a as novel constituents of fibrotic liver[J].Journal of Proteome Research,2012,11(8):4052-4064.
[19]刘玉,冯立波,钟武.心肌梗死与wnt信号通路的相关性研究现状[J].重庆医学.,2017,46(6):845-847.
[20]Rashid S T,Humphries J D,Byron A,et al.Proteomic analysis of extracellular matrix from the hepatic stellate cell line LX-2 identifies CYR61 and Wnt-5a as novel constituents of fibrotic liver[J].Journal of Proteome Research,2012,11(8):4052-4064.
[21]Koval A,Purvanov V,Egger-Adam D,et al.Yellow submarine of the Wnt/Frizzled signaling:Submerging from the G protein harbor to the targets[J].Biochemical Pharmacology,2011,82(10):1311-1319.
[22]Ozhan G,Weidinger G.Wnt/β-catenin signaling in heart regeneration[J].Cell Regeneration,2015,4(1).
[23]Klaus A,Muller M,Schulz H,et al.Wnt/-catenin and Bmp signals control distinct sets of transcription factors in cardiac progenitor cells[J].Proceedings of the National Academy of Sciences,2012,109(27):10921-10926.
[24]Palevski D,Levin Kotler L P,Kain D,et al.Loss of Macrophage Wnt Secretion Improves Remodeling and Function After Myocardial Infarction in Mice[J].Journal of the American Heart Association,2017,6(1):e4387.
[25]Liu J,Wu X,Mitchell B,et al.A Small-Molecule Agonist of the Wnt Signaling Pathway[J].Angewandte Chemie International Edition,2005,44(13):1987-1990.
[26]Kuncewitch M,Yang W,Jacob A,et al.Stimulation of Wnt/β-catenin signaling pathway with Wnt agonist reduces organ injury after hemorrhagic shock[J].Journal of Trauma and Acute Care Surgery,2015,78(4):793-800.
[27]Bastakoty D,Saraswati S,Joshi P,et al.Temporary,Systemic Inhibition of the WNT/beta-Catenin Pathway promotes Regenerative Cardiac Repair following Myocardial Infarct[J].Cell Stem Cells Regen Med,2016,2(2).
[28]Tao J,Abudoukelimu M,Ma Y,et al.Secreted frizzled related protein 1protects H9C2 cells from hypoxia/re-oxygenation injury by blocking the Wnt signaling pathway[J].Lipids in Health and Disease,2016,15(1).
[29]Ming M,Wang S,Wu W,et al.Activation of Wnt/β-Catenin Protein Signaling Induces Mitochondria-mediated Apoptosis in Hematopoietic Progenitor Cells[J].Journal of Biological Chemistry,2012,287(27):22683-22690.
[30]Zhang Z,Deb A,Zhang Z,et al.Secreted frizzled related protein 2protects cells from apoptosis by blocking the effect of canonical Wnt3a[J].Journal of Molecular and Cellular Cardiology,2009,46(3):370-377.
[31]Korkmaz-Ic?z S,Radovits T,SzabóG.Targeting phosphodiesterase 5as a therapeutic option against myocardial ischaemia/reperfusion injury and for treating heart failure[J].British Journal of Pharmacology,2017.Epub ahead of print
[32]Agrawal V,Gupta J K,Qureshi S S,et al.Role of cardiac renin angiotensin system in ischemia reperfusion injury and preconditioning of heart[J].Indian Heart Journal,2016,68(6):856-861.
[33]Lin J C,Kuo W W,Baskaran R,et al.Enhancement of beta-catenin in cardiomyocytes suppresses survival protein expression but promotes apoptosis and fibrosis[J].Cardiol J,2016.
[34]Kim K,Pang K M,Evans M,et al.Overexpression of beta-catenin induces apoptosis independent of its transactivation function with LEF-1 or the involvement of major G1 cell cycle regulators[J].Mol Biol Cell,2000,11(10):3509-3523.
[35]Prensner J R,Chen W,Han S,et al.The Long Non-Coding RNA PCAT-1 Promotes Prostate Cancer Cell Proliferation through c Myc[J].Neoplasia,2014,16(11):900-908.
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