Hei Bingchang, Zhang Anzhe, Yang Meiming, Jiang Yunfei, Guan Zhanjiang
Intensive Care Unit, The Third Affiliated Hospital of Qiqihar Medical University, Qiqihar, 161099, China.
Respiratory Medicine, Tailai County People's Hospital, Qiqihar, 162400, China.
Biochem Genet. 2024 Dec 25. doi: 10.1007/s10528-024-11007-w.
Pulmonary hypertension (PH) is a progressive disease characterized by vascular reHypoxiaing, endothelial cell dysfunction, and inflammation. Liver Kinase B1 (LKB1, also known as STK11) is a central regulator of cell polarity and energy homeostasis. However, its specific role and mechanism of action in PH remain unclear. Human pulmonary arterial endothelial cells (hPAECs) were cultured in vitro to establish PH cell Hypoxias under normoxic and hypoxic conditions. The expression of LKB1 was detected by reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR) and western blotting, and its effect on hPAECs function was investigated by overexpression and inhibition of LKB1. Furthermore, cell proliferation was assessed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, apoptosis was measured by flow cytometry, inflammatory cytokine secretion was evaluated using enzyme-linked immunosorbent assay (ELISA), and the expression of AMP-activated protein kinase (AMPK) signaling pathway-related proteins was analyzed by western blotting. LKB1 expression was significantly reduced in hypoxia-treated hPAECs compared with that in normoxic controls, and LKB1 overexpression significantly ameliorated the hypoxia-induced decrease in cell proliferation and increase in apoptosis as well as inflammatory factor secretion. The AMPK agonist (GSK621) reversed the dysfunction caused by LKB1 inhibition, indicating that LKB1 regulates hPAECs function through the AMPK signaling pathway. LKB1 plays a protective role in PH by inhibiting hPAECs dysfunction via activation of the AMPK pathway.
肺动脉高压(PH)是一种以血管重塑、内皮细胞功能障碍和炎症为特征的进行性疾病。肝脏激酶B1(LKB1,也称为STK11)是细胞极性和能量稳态的核心调节因子。然而,其在PH中的具体作用和作用机制仍不清楚。体外培养人肺动脉内皮细胞(hPAECs),在常氧和缺氧条件下建立PH细胞模型。通过逆转录定量聚合酶链反应(RT-qPCR)和蛋白质印迹法检测LKB1的表达,并通过过表达和抑制LKB1来研究其对hPAECs功能的影响。此外,使用3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐(MTT)法评估细胞增殖,通过流式细胞术检测细胞凋亡,使用酶联免疫吸附测定(ELISA)评估炎性细胞因子分泌,并通过蛋白质印迹法分析AMP活化蛋白激酶(AMPK)信号通路相关蛋白的表达。与常氧对照组相比,缺氧处理的hPAECs中LKB1表达显著降低,LKB1过表达显著改善了缺氧诱导的细胞增殖减少、凋亡增加以及炎性因子分泌增加。AMPK激动剂(GSK621)逆转了LKB1抑制引起的功能障碍,表明LKB1通过AMPK信号通路调节hPAECs功能。LKB1通过激活AMPK途径抑制hPAECs功能障碍,在PH中发挥保护作用。