Department of Cardiology, The Affiliated Huaian No. 1 People's Hospital of Nanjing Medical University, Huai'an, Jiangsu, China (mainland).
Med Sci Monit. 2019 Jul 22;25:5426-5434. doi: 10.12659/MSM.915108.
BACKGROUND The neutrophil inflammatory protein, lipocalin-2 (NGAL), is elevated in certain forms of cardiac hypertrophy and acute heart failure. However, the specific role of NGAL in cardiac hypoxia injury is unclear. This study aimed to elucidate the functional role of NGAL in cardiomyocyte hypoxia injury. MATERIAL AND METHODS Neonatal rat cardiomyocytes were transfected with adenovirus [(Ad-NGAL] to overexpress human-NGAL and then were exposed to hypoxia for 24 h to establish a hypoxia model. Cell inflammation was detected by RT-PCT and ELISA assay. Cell apoptosis was detected by TUNEL assay. Oxidative stress was also detected by commercial kits. RESULTS An increased inflammatory response, apoptosis, and augmented oxidative stress were observed after exposure to hypoxia, while NGAL overexpression in cells increased the expression and release of inflammatory cytokines. NGAL overexpression also increased the number of apoptotic cells and the imbalance of Bax/Bcl-2 protein expression. Moreover, NGAL overexpression increased the levels of reactive oxygen species and oxidase activity, but reduced anti-oxidase activity. Mechanistically, we found that NGAL decreased the expression of integrin ß3, but not the expression of integrin avß3 and avß5, thus inhibiting the downstream protein AKT. When we used the constitutively activated AKT overexpression adenovirus to activate AKT, the deteriorated phenotype by NGAL was counteracted. CONCLUSIONS NGAL can directly affect cardiomyocytes and cause cardiomyocyte deteriorated hypoxia injury through inhibiting integrin ß3 signaling.
中性粒细胞炎症蛋白脂联素-2(NGAL)在某些形式的心脏肥大和急性心力衰竭中升高。然而,NGAL 在心脏缺氧损伤中的具体作用尚不清楚。本研究旨在阐明 NGAL 在心肌细胞缺氧损伤中的功能作用。
用腺病毒[(Ad-NGAL)]转染新生大鼠心肌细胞以过表达人-NGAL,然后将其暴露于缺氧 24 小时以建立缺氧模型。通过 RT-PCT 和 ELISA 检测细胞炎症。通过 TUNEL 检测细胞凋亡。通过商业试剂盒检测氧化应激。
暴露于缺氧后观察到炎症反应、细胞凋亡和氧化应激增强增加,而细胞中 NGAL 的过表达增加了炎症细胞因子的表达和释放。NGAL 的过表达还增加了凋亡细胞的数量和 Bax/Bcl-2 蛋白表达的失衡。此外,NGAL 的过表达增加了活性氧的水平和氧化酶的活性,但降低了抗氧化酶的活性。从机制上讲,我们发现 NGAL 降低了整合素 β3 的表达,但不降低整合素 avβ3 和 avβ5 的表达,从而抑制下游蛋白 AKT。当我们使用组成型激活的 AKT 过表达腺病毒激活 AKT 时,NGAL 引起的恶化表型被抵消。
NGAL 可直接影响心肌细胞,并通过抑制整合素 β3 信号导致心肌细胞恶化的缺氧损伤。