Division of Cardiology, Shin Kong Wu Ho-Su Memorial Hospital, and School of Medicine, Fu-Jen Catholic University, New Taipei City, Taipei, Taiwan.
J Biomed Sci. 2011 May 4;18(1):27. doi: 10.1186/1423-0127-18-27.
Visfatin, a adipocytokine with insulin-mimetic effect, plays a role in endothelial angiogenesis. Hyperbaric oxygen (HBO) has been used in medical practice. However, the molecular mechanism of beneficial effects of HBO is poorly understood. We sought to investigate the cellular and molecular mechanisms of regulation of visfatin by HBO in human coronary arterial endothelial cells (CAECs).
Human CAECs were exposed to 2.5 atmosphere absolute (ATA) of oxygen in a hyperbaric chamber. Western blot, real-time polymerase chain reaction, and promoter activity assay were performed. In vitro glucose uptake and tube formation was detected.
Visfatin protein (2.55-fold) and mRNA (2.53-fold) expression were significantly increased after exposure to 2.5 ATA HBO for 4 to 6 h. Addition of SP600125 and JNK siRNA 30 min before HBO inhibited the induction of visfatin protein. HBO also significantly increased DNA-protein binding activity of AP-1 and visfatin promoter activity. Addition of SP600125 and TNF-α monoclonal antibody 30 min before HBO abolished the DNA-protein binding activity and visfatin promoter activity induced by HBO. HBO significantly increased secretion of TNF-α from cultured human CAECs. Exogenous addition of TNF-α significantly increased visfatin protein expression while TNF-α antibody and TNF-α receptor antibody blocked the induction of visfatin protein expression induced by HBO. HBO increased glucose uptake in human CAECs as HBO and visfatin siRNA and TNF-α antibody attenuated the glucose uptake induced by HBO. HBO significantly increased the tube formation of human CAECs while visfatin siRNA, TNF-α antibody inhibited the tube formation induced by HBO.
HBO activates visfatin expression in cultured human CAECs. HBO-induced visfatin is mediated by TNF-α and at least in part through JNK pathway.
脂联素是一种具有胰岛素样作用的脂肪细胞因子，在血管内皮生成中起作用。高压氧（HBO）已在医学实践中应用。然而，HBO 有益作用的分子机制尚不清楚。我们试图研究 HBO 对人冠状动脉内皮细胞（CAEC）中脂联素的调节的细胞和分子机制。
人 CAEC 在高压舱中暴露于 2.5 个大气压的氧气下。进行 Western blot、实时聚合酶链反应和启动子活性测定。检测体外葡萄糖摄取和管形成。
暴露于 2.5ATA HBO 4 至 6 小时后，脂联素蛋白（增加 2.55 倍）和 mRNA（增加 2.53 倍）表达显著增加。在 HBO 之前 30 分钟添加 SP600125 和 JNK siRNA 抑制脂联素蛋白的诱导。HBO 还显著增加了 AP-1 的 DNA-蛋白结合活性和脂联素启动子活性。在 HBO 之前 30 分钟添加 SP600125 和 TNF-α 单克隆抗体可消除 HBO 诱导的 DNA-蛋白结合活性和脂联素启动子活性。HBO 显著增加培养的人 CAEC 中 TNF-α 的分泌。外源性添加 TNF-α 显著增加脂联素蛋白表达，而 TNF-α 抗体和 TNF-α 受体抗体阻断了 HBO 诱导的脂联素蛋白表达的诱导。HBO 增加了人 CAEC 的葡萄糖摄取，而 HBO 和脂联素 siRNA 以及 TNF-α 抗体减弱了 HBO 诱导的葡萄糖摄取。HBO 显著增加了人 CAEC 的管形成，而脂联素 siRNA 和 TNF-α 抗体抑制了 HBO 诱导的管形成。
HBO 激活培养的人 CAEC 中的脂联素表达。HBO 诱导的脂联素是由 TNF-α介导的，至少部分是通过 JNK 途径。
