Shraga Segal Department of Microbiology, Immunology and Genetics, Faculty of Health Sciences, Ben-Gurion University of the Negev, Beer-Sheva 8410501, Israel.
Regenerative Medicine & Stem Cell Research Center, Ben-Gurion University of the Negev, Beer-Sheva 8410501, Israel.
Int J Mol Sci. 2022 Sep 7;23(18):10272. doi: 10.3390/ijms231810272.
Cardiovascular morbidity is the leading cause of death of obstructive sleep apnea (OSA) syndrome patients. Nocturnal airway obstruction is associated with intermittent hypoxia (IH). In our previous work with cell lines, incubation with sera from OSA patients induced changes in cell morphology, NF-κB activation and decreased viability. A decrease in beating rate, contraction amplitude and a reduction in intracellular calcium signaling was also observed in human cardiomyocytes differentiated from human embryonic stem cells (hESC-CMs). We expanded these observations using a new controlled IH in vitro system on beating hESC-CMs. The Oxy-Cycler system was programed to generate IH cycles. Following IH, we detected the activation of Hif-1α as an indicator of hypoxia and nuclear NF-κB p65 and p50 subunits, representing pro-inflammatory activity. We also detected the secretion of inflammatory cytokines, such as MIF, PAI-1, MCP-1 and CXCL1, and demonstrated a decrease in beating rate of hESC-CMs following IH. IH induces the co-activation of inflammatory features together with cardiomyocyte alterations which are consistent with myocardial damage in OSA. This study provides an innovative approach for in vitro studies of OSA cardiovascular morbidity and supports the search for new pharmacological agents and molecular targets to improve diagnosis and treatment of patients.
心血管疾病是阻塞性睡眠呼吸暂停(OSA)综合征患者的主要死亡原因。夜间气道阻塞与间歇性低氧(IH)有关。在我们之前与细胞系的合作中,孵育 OSA 患者的血清会导致细胞形态、NF-κB 激活和活力降低的变化。在从人胚胎干细胞(hESC-CMs)分化而来的人心肌细胞中,还观察到心率下降、收缩幅度减小和细胞内钙信号转导减少。我们使用新的体外搏动 hESC-CMs 受控 IH 系统扩展了这些观察结果。Oxy-Cycler 系统被编程以产生 IH 循环。在 IH 之后,我们检测到 Hif-1α 的激活作为缺氧的指标,以及核 NF-κB p65 和 p50 亚基,代表炎症活性。我们还检测到炎症细胞因子的分泌,如 MIF、PAI-1、MCP-1 和 CXCL1,并证明 IH 后 hESC-CMs 的心率下降。IH 诱导炎症特征的共同激活以及心肌细胞的改变,与 OSA 中的心肌损伤一致。这项研究为 OSA 心血管疾病的体外研究提供了一种创新方法,并支持寻找新的药物和分子靶点,以改善患者的诊断和治疗。