胡洸瑜, 杨容金, 孙芳芳, 宋丹, 郭兰燕, 张玲, 郭雄, 张富洋, 陈希瑶. 过表达Fas激活的丝氨酸/苏氨酸激酶减轻缺氧/复氧损伤导致的心肌细胞线粒体呼吸功能障碍[J]. 心脏杂志, 2021, 33(2): 123-126, 132. DOI: 10.12125/j.chj.202012042
    引用本文: 胡洸瑜, 杨容金, 孙芳芳, 宋丹, 郭兰燕, 张玲, 郭雄, 张富洋, 陈希瑶. 过表达Fas激活的丝氨酸/苏氨酸激酶减轻缺氧/复氧损伤导致的心肌细胞线粒体呼吸功能障碍[J]. 心脏杂志, 2021, 33(2): 123-126, 132. DOI: 10.12125/j.chj.202012042
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    Guang-yu HU, Rong-jin YANG, Fang-fang SUN, Dan SONG, Lan-yan GUO , Ling ZHANG, Xiong GUO, Fu-yang ZHANG, Xi-yao CHEN. Overexpression of Fas-activated serine/threonine kinase ameliorates hypoxia/reoxygenation injury-induced mitochondrial respiratory dysfunction in cardiomyocytes[J]. Chinese Heart Journal, 2021, 33(2): 123-126, 132. DOI: 10.12125/j.chj.202012042
    Citation: Guang-yu HU, Rong-jin YANG, Fang-fang SUN, Dan SONG, Lan-yan GUO , Ling ZHANG, Xiong GUO, Fu-yang ZHANG, Xi-yao CHEN. Overexpression of Fas-activated serine/threonine kinase ameliorates hypoxia/reoxygenation injury-induced mitochondrial respiratory dysfunction in cardiomyocytes[J]. Chinese Heart Journal, 2021, 33(2): 123-126, 132. DOI: 10.12125/j.chj.202012042
    shu

    过表达Fas激活的丝氨酸/苏氨酸激酶减轻缺氧/复氧损伤导致的心肌细胞线粒体呼吸功能障碍

    Overexpression of Fas-activated serine/threonine kinase ameliorates hypoxia/reoxygenation injury-induced mitochondrial respiratory dysfunction in cardiomyocytes

      摘要
    • 摘要:
        目的  研究Fas激活的丝氨酸/苏氨酸激酶（Fas-activated serine/threonine kinase，FASTK）在缺氧/复氧（hypoxia/reoxygenation, H/R）损伤导致的心肌细胞线粒体呼吸功能障碍中的作用。
        方法  分离(1−2)d的野生C57BL/6小鼠原代心肌细胞并进行体外培养。转染对照或FASTK过表达的腺病毒后，心肌细胞接受H/R损伤。分别采用Western blot和RT-PCR检测FASTK蛋白和mRNA表达水平，采用Seahorse能量分析仪检测心肌细胞线粒体呼吸功能，采用caspase-3活性检测试剂盒、乳酸脱氢酶（lactate dehydrogenase，LDH）活性检测试剂盒、MTT细胞活力检测试剂盒评估心肌细胞存活情况。
        结果  较常氧组，H/R损伤导致心肌细胞FASTK蛋白和mRNA表达显著降低（均P<0.01）。转染FASTK过表达腺病毒可显著上调心肌细胞FASTK蛋白和mRNA水平（均P<0.01）。H/R损伤抑制心肌细胞基础和最大线粒体氧耗率并降低三磷酸腺苷（adenosine triphosphate，ATP）生成速率（均P<0.01），而FASTK过表达可显著减轻H/R导致的线粒体呼吸功能障碍（均P<0.001）。FASTK过表达抑制了caspase-3活化，也显著减轻了H/R损伤导致的心肌细胞死亡和LDH释放（均P<0.001）。
        结论  过表达FASTK可显著减轻H/R损伤导致的心肌细胞线粒体呼吸功能障碍。

       

      Abstract:
        AIM  The study aims to investigate the role of Fas-activated serine/threonine kinase (FASTK) in hypoxia/reoxygenation (H/R)-induced mitochondrial respiratory dysfunction in cardiomyocytes.
        METHODS  Primary cardiomyocytes were isolated from 1 to 2-day-old C57BL6/J mice and were in vitro cultured. Cardiomyocytes were transfected with adenovirus vectors carrying control green fluorescent protein (Ad-Control) or full-length FASTK (Ad-FASTK). Mitochondrial respiratory function was evaluated by the Seahorse metabolic analyzer. Cardiomyocyte death was evaluated by caspase-3 activity, lactate dehydrogenase (LDH) release, and MTT cell viability assay.
        RESULTS  H/R obviously suppressed FASTK mRNA and protein expression in cardiomyocytes (all P<0.01). Compared with Ad-Control, Ad-FASTK significantly increased FASTK mRNA and protein expression levels. Overexpression of FASTK significantly preserved mitochondrial respiratory function in H/R-exposed cardiomyocytes, as evidenced by higher basal and maximal respiratory capability as well as stronger adenosine triphosphate (ATP) production (all P<0.01). Moreover, FASTK overexpression ameliorated H/R-induced caspase-3 activation, LDH release, and cardiomyocyte death (all P<0.01).
        CONCLUSION  Overexpression of FASTK ameliorates H/R-associated mitochondrial respiratory dysfunction and cell death in cardiomyocytes.

       

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