Hemichannel-mediated volume regulation contributes to IPC-induced cardiomyocyte protection
Hemichannel-mediated volume regulation contributes to IPC-induced cardiomyocyte protection
复制标题
半通道介导的容量调节有助于 IPC 诱导的心肌细胞保护
DOI：
10.3892/etm.2018.7127
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发表时间：
2019
影响因子：
2.7
通讯作者：
Luo Yukun
中科院分区：
文献类型：
--
作者：
Wang Wenying;Zheng Dedong;Li Huiya;Huang Jinhua;Chen Huijun;Ying Teng;Fang Jun;Luo Yukun
Cx43参与缺血预处理（IPC）。然而，Cx43形成的半通道在IPC中的参与和潜在的潜在机制仍不清楚。本研究通过观察IPC对心肌细胞容积的调节，探讨半通道在IPC心肌保护中的作用。在这项研究中，小鼠心肌细胞分别用半通道阻断剂，辛醇或18 α-甘草酸（18 α-GA）和Cx43沉默的慢病毒处理。随后在低渗溶液中培养以模拟缺血再灌注（SIR）和全身缺血预处理（SIP）。在加入低渗溶液后30 min，通过倒置显微镜检测细胞形态和体积（面积）变化。台盼蓝染色法检测心肌细胞死亡率。分析显示，无论治疗如何，低渗溶液都会加重细胞水肿：与初始条件相比，（加入溶液前的时刻，0 min），30 min后体积面积显著增加（对于低渗+DMSO，分别为5，050 ± 1，511和3，464 ±723 µm2;对于低渗+乱序慢病毒载体，分别为5，517 ± 1，128和2，331 ±536 µm2; P<0.05）。当在低渗添加后30分钟与0分钟之间进行比较时，任一治疗减轻了水肿状况（对于低渗+辛醇，2，990 ±765 vs. 2，821 ±773 µm2;对于低渗+18 a-GA，4，817 ± 1，306 vs. 4，762 ± 1，271 µm2;对于低渗+Cx43-沉默，3，627 ±688 vs. 3，419 ±814 µm2; P>0.05）。值得注意的是，结果表明SIP组与SIR对应组相比具有较低的死亡率;低渗+辛醇、低渗+18 α-GA和低渗+Cx43沉默组与其各自的对照组相比显示出显著降低的死亡率（分别为35.70±1.02、30.76±2.20和53.58±2.14%; 30.89±2.37和54.12±2.55%; P均<0.05）。结果提示，缺血预处理可能通过阻断半通道的开放进而介导心肌细胞的容积调节而提供心肌保护作用。
Cx43 has been documented to be involved in ischemic preconditioning (IPC). However, the participation of Cx43-formed hemichannels in IPC and the potential underlying mechanisms remain unclear. The present study focused on cardiomyocytes' volume regulation during IPC to investigate the role of hemichannels in the IPC-induced cardioprotection. In the study, mice cardiomyocytes were respectively treated with a hemichannel blocker, octanol or 18a-Glycyrrhizic acid (18a-GA), and a Cx43-silenced lentivirus. They were subsequently cultured in hypotonic solution to simulate ischemic reperfusion (SIR) and systemic ischemic preconditioning (SIP). Cell morphology and volumetric (area) change were detected by inverted microscopy at 30 min following the addition of hypotonic solution. Cardiomyocyte mortality was assessed by trypan blue stain assay. The analyses revealed that regardless of the treatments, hypotonic solution aggravated cell edema: Compared with the initial condition (the moment before the solution addition, 0 min), the volumetric area increased significantly 30 min later (for hypotonic+DMSO, 5,050±1,511 vs. 3,464±723 µm2; for hypotonic+scramble lentiviral vector, 5,517±1,128 vs. 2,331±536 µm2; P<0.05, respectively). Either treatment alleviated the edematous condition when a comparison was made between 30 min after the hypotonic addition and 0 min (for hypotonic+octanol, 2,990±765 vs. 2,821±773 µm2; for hypotonic+18a-GA, 4,817±1,306 vs. 4,762±1,271 µm2; for hypotonic+Cx43-silenced, 3,627±688 vs. 3,419±814 µm2; P>0.05 for all). Notably, results indicated that the SIP group had lower mortality rates compared with its SIR counterpart; the hypotonic+octanol, hypotonic+18a-GA, and hypotonic+Cx43-silenced group showed markedly-declined mortality when compared with their respective control groups (respectively, 35.70±1.02, 30.76±2.20 vs. 53.58±2.14%; 30.89±2.37 vs. 54.12±2.55%; P<0.05 for all). The results suggest that ischemic preconditioning may provide cardioprotection by blocking the opening of the hemichannels and further mediating the volume regulation of cardiomyocytes.