Department of Anesthesiology. First Affiliated Hospital of Hebei North College, Zhangjiakou, China.
Bioengineered. 2022 Apr;13(4):9369-9386. doi: 10.1080/21655979.2022.2054762.
Myocardial hypoxia/reoxygenation (H/R) injury is a common pathological change in patients with acute myocardial infarction undergoing reperfusion therapy. Dexmedetomidine (DEX) has been found to substantially improve ischemia-mediated cell damage. Here, we focus on probing the role and mechanism of DEX in ameliorating myocardial H/R injury. Oxygen-glucose deprivation and reoxygenation (OGD/R) were applied to construct the H/R injury model in human myocardial cell lines. After different concentrations of DEX's treatment, cell counting kit-8 (CCK-8) assay and BrdU assay were employed to test cell viability. The profiles of apoptosis-related proteins Bcl2, Bax, Bad and Caspase3, 8, 9 were determined by Western blot (WB). The expression of inflammatory factors interleukin 1β (IL-1β) and tumor necrosis factor-α (TNF-α) was checked by reverse transcription-polymerase chain reaction (RT-PCR). By conducting WB, we examined the expression of NF-κB, Sirt1, Tet methylcytosine dioxygenase 1 (TET1) and DNA methylation-related proteins (DNA methyltransferase 1, DNMT1; DNA methyltransferase 3 alpha, DNMT3A; and DNA methyltransferase 3 beta, DNMT3B). Our data showed that OGD/R stimulation distinctly hampered the viability and elevated apoptosis and inflammatory factor expression in cardiomyocytes. DEX treatment notably impeded myocardial apoptosis and inflammation and enhanced cardiomyocyte viability. OGD/R enhanced total DNA methylation levels in cardiomyocytes, while DEX curbed DNA methylation. In terms of mechanism, inhibiting TET1 or Sirtuin1 (Sirt1) curbed the DEX-mediated myocardial protection. TET1 strengthened demethylation of the Sirt1 promoter and up-regulated Sirt1. DEX up-regulates Sirt1 by accelerating TET1 and mediating demethylation of the Sirt1 promoter and improves H/R-mediated myocardial injury.
心肌缺氧/复氧(H/R)损伤是急性心肌梗死患者再灌注治疗中常见的病理变化。右美托咪定(DEX)已被发现可显著改善缺血介导的细胞损伤。在这里,我们重点探讨 DEX 改善心肌 H/R 损伤的作用和机制。采用氧葡萄糖剥夺和再氧合(OGD/R)构建人心肌细胞系 H/R 损伤模型。用不同浓度的 DEX 处理后,采用细胞计数试剂盒-8(CCK-8)法和 BrdU 法检测细胞活力。用 Western blot(WB)法测定凋亡相关蛋白 Bcl2、Bax、Bad 和 Caspase3、8、9 的表达。采用逆转录-聚合酶链反应(RT-PCR)法检测炎症因子白细胞介素 1β(IL-1β)和肿瘤坏死因子-α(TNF-α)的表达。通过 WB,我们检测了 NF-κB、Sirt1、Tet 甲基胞嘧啶双加氧酶 1(TET1)和 DNA 甲基化相关蛋白(DNA 甲基转移酶 1、DNMT1;DNA 甲基转移酶 3α、DNMT3A;和 DNA 甲基转移酶 3β、DNMT3B)的表达。我们的数据表明,OGD/R 刺激明显抑制了心肌细胞的活力,并升高了凋亡和炎症因子的表达。DEX 处理明显抑制心肌细胞凋亡和炎症,增强心肌细胞活力。OGD/R 增强了心肌细胞的总 DNA 甲基化水平,而 DEX 抑制了 DNA 甲基化。就机制而言,抑制 TET1 或 Sirtuin1(Sirt1)抑制了 DEX 介导的心肌保护。TET1 增强了 Sirt1 启动子的去甲基化并上调了 Sirt1。DEX 通过加速 TET1 并介导 Sirt1 启动子的去甲基化而上调 Sirt1,改善 H/R 介导的心肌损伤。