Chen Manli, Liu Qin, Chen Lijian, Zhang Lei, Gu Erwei
Department of Anesthesiology, The First Affiliated Hospital, Medical School of Zhejiang University, Hangzhou, China.
Department of Anesthesiology, The First Affiliated Hospital of Anhui Medical University, No. 218 Jixi Road, Hefei, Anhui, China.
Apoptosis. 2017 May;22(5):662-671. doi: 10.1007/s10495-017-1347-5.
Remifentanil postconditioning (RPC) elicits cardioprotection against ischemia/reperfusion injury (IRI) by attenuating apoptosis associated with endoplasmic reticulum stress (ERS). Histone H3, acetylation modifications of histone H3, and histone deacetylases (HDAC) also have key roles in the mediation of the survival and apoptosis of cardiomyocytes. In this study, an in vitro IRI model was established with H9c2 cardiomyoblasts to investigate the role of histone H3 acetylation and HDAC3 in RPC-induced attenuation of ERS-associated apoptosis. Briefly, H9c2 cardiomyoblasts were randomly subjected to hypoxia/reoxygenation with and without remifentanil administered at the onset of reoxygenation. Results showed that RPC increased cell viability and prevented cell apoptosis (evidenced by CCK-8 cell viability assays and flow cytometry), and these effects were accompanied by lower levels of expression of GRP78, CHOP, cleaved caspase-12, and cleaved caspase-3. RPC also mimicked the effects of SAHA by increasing the amount of histone H3 deacetylation and decreasing up-regulation of HDAC at both the mRNA and protein levels in response to HR. Finally, RPC-induced protective effects against HR, including attenuation of ERS-associated protein markers, deacetylation of histone H3, and down-regulation of HDAC3 were completely abolished by pretreatment with thapsigargin (TG, a specific ERS activator). In contrast, these effects were not found to be enhanced after pretreatment with 4-phenyl butyric acid (4-PBA, a widely used ERS inhibitor). The present results demonstrate that RPC protects H9c2 cardiomyoblasts from HR injury, and this protection involves an attenuation of ERS-associated apoptosis, which mediates a reduction in HDAC3 expression and an increase in histone H3 deacetylation.
瑞芬太尼后处理（RPC）通过减轻与内质网应激（ERS）相关的细胞凋亡，对缺血/再灌注损伤（IRI）发挥心脏保护作用。组蛋白H3、组蛋白H3的乙酰化修饰以及组蛋白去乙酰化酶（HDAC）在心肌细胞存活和凋亡的介导中也起着关键作用。在本研究中，利用H9c2心肌母细胞建立体外IRI模型，以研究组蛋白H3乙酰化和HDAC3在RPC诱导减轻ERS相关凋亡中的作用。简要地说，H9c2心肌母细胞在复氧开始时随机接受有无瑞芬太尼的缺氧/复氧处理。结果显示，RPC提高了细胞活力并防止细胞凋亡（CCK-8细胞活力测定和流式细胞术证明），并且这些作用伴随着GRP78、CHOP、裂解的caspase-12和裂解的caspase-3表达水平降低。RPC还通过增加组蛋白H3去乙酰化量并在mRNA和蛋白质水平上降低HR诱导的HDAC上调，模拟了SAHA的作用。最后，用毒胡萝卜素（TG，一种特异性ERS激活剂）预处理完全消除了RPC诱导的对HR的保护作用，包括减轻ERS相关蛋白标志物、组蛋白H3去乙酰化以及HDAC3下调。相反，用4-苯基丁酸（4-PBA，一种广泛使用的ERS抑制剂）预处理后未发现这些作用增强。本研究结果表明，RPC保护H9c2心肌母细胞免受HR损伤，且这种保护涉及减轻ERS相关凋亡，其介导HDAC3表达减少和组蛋白H3去乙酰化增加。
