Department of Anaesthesia, General Intensive Care and Pain Management, Medical University of Vienna, Vienna, Austria.
Department of Anaesthesiology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
Shock. 2018 May;49(5):556-563. doi: 10.1097/SHK.0000000000000998.
The noble gas argon induces cardioprotection in a rabbit model of myocardial ischemia and reperfusion. However, no studies in human primary cells or subjects have been performed so far. We used human cardiac myocyte-like progenitor cells (HCMs) to investigate the protective effect on the cellular level.
HCMs were pretreated with 30% or 50% argon before oxygen-glucose deprivation (OGD) and reperfusion. We evaluated apoptotic states by flow cytometry and the activation of mitogen-activated protein kinase (MAPKs) members extracellular signal-regulated kinase (ERK), c-jun N-terminal kinase (JNK), p38 MAPkinase, and protein kinase B (Akt) by Westernblot analysis and by activity assays of downstream transcription factors. Specific inhibitors were used to proof a significant participation of these pathways in the protection by argon. Beneficial effects were further assessed by TdT-mediated dUTP-biotin nick end labeling (TUNEL) assay, lactate dehydrogenase (LDH), mitochondrial deoxyribonucleic acid (mtDNA), and cytokine release.
Pretreatment with 30% or 50% argon for 90 min before OGD resulted in a significant protection of HCMs against apoptosis. This effect was reversed by the application of MAPK and Akt inhibitors during argon exposure. Argon 30% reduced the release of LDH by 33% and mtDNA by 45%. The release of interleukin 1β was reduced by 44% after OGD and more than 90% during reperfusion.
Pretreatment with argon protects HCMs from apoptosis under ischemic conditions via activation of Akt, Erk, and biphasic regulation of JNK. Argon gas is cheap and easily administrable, and might be a novel therapy to reduce myocardial ischemia-reperfusion injury.
稀有气体氩气可诱导兔心肌缺血再灌注模型中的心脏保护作用。然而,目前尚未在人体原代细胞或受试者中进行研究。我们使用人心肌细胞样祖细胞(HCM)来研究其在细胞水平上的保护作用。
在氧葡萄糖剥夺(OGD)和再灌注之前,用 30%或 50%氩气预处理 HCM。我们通过流式细胞术评估细胞凋亡状态,并通过 Westernblot 分析和下游转录因子的活性测定来评估丝裂原激活的蛋白激酶(MAPKs)成员细胞外信号调节激酶(ERK)、c-jun N 末端激酶(JNK)、p38 MAPkinase 和蛋白激酶 B(Akt)的激活。使用特异性抑制剂证明这些途径在氩气保护中的显著参与。通过 TdT 介导的 dUTP-生物素缺口末端标记(TUNEL)测定、乳酸脱氢酶(LDH)、线粒体脱氧核糖核酸(mtDNA)和细胞因子释放进一步评估有益作用。
在 OGD 前用 30%或 50%氩气预处理 90 分钟可显著保护 HCM 免受细胞凋亡。在氩气暴露期间应用 MAPK 和 Akt 抑制剂可逆转此作用。氩气 30%可使 LDH 释放减少 33%,mtDNA 释放减少 45%。OGD 后,白细胞介素 1β的释放减少了 44%,再灌注期间则减少了 90%以上。
在缺血条件下,用氩气预处理可通过激活 Akt、Erk 和 JNK 的双相调节来保护 HCM 免受细胞凋亡。氩气廉价且易于管理,可能是减少心肌缺血再灌注损伤的新疗法。