Department of Medical Pharmacology and Physiology, University of Missouri, Columbia, Missouri 65212, USA.
Am J Physiol Heart Circ Physiol. 2011 Jan;300(1):H84-93. doi: 10.1152/ajpheart.00749.2010. Epub 2010 Oct 29.
The present study determined whether AMP-activated protein kinase (AMPK) regulates heme oxygenase (HO)-1 gene expression in endothelial cells (ECs) and if HO-1 contributes to the biological actions of this kinase. Treatment of human ECs with the AMPK activator 5-aminoimidazole-4-carboxamide-1-β-d-ribofuranoside (AICAR) stimulated a concentration- and time-dependent increase in HO-1 protein and mRNA expression that was associated with a prominent increase in nuclear factor-erythroid 2-related factor 2 (Nrf2) protein. Induction of HO-1 was also observed in rat carotid arteries after the in vivo application of AICAR. Induction of HO-1 by AICAR was blocked by the AMPK inhibitor compound C, the adenosine kinase inhibitor 5'-iodotubercidin, and by silencing AMPK-α(1/2) and was mimicked by the AMPK activator A-769662 and by infecting ECs with an adenovirus expressing constitutively active AMPK-α(1). AICAR also induced a significant rise in HO-1 promoter activity that was abolished by mutating the antioxidant responsive elements of the HO-1 promoter or by the overexpression of dominant negative Nrf2. Finally, activation of AMPK inhibited cytokine-mediated EC death, and this was prevented by the HO inhibitor tin protoporphyrin-IX or by silencing HO-1 expression. In conclusion, AMPK stimulates HO-1 gene expression in human ECs via the Nrf2/antioxidant responsive element signaling pathway. The induction of HO-1 mediates the antiapoptotic effect of AMPK, and this may provide an important adaptive response to preserve EC viability during periods of metabolic stress.
本研究旨在确定 AMP 激活的蛋白激酶(AMPK)是否调节内皮细胞(ECs)中的血红素加氧酶(HO)-1 基因表达,以及 HO-1 是否有助于该激酶的生物学作用。用 AMPK 激活剂 5-氨基咪唑-4-甲酰胺-1-β-D-呋喃核糖苷(AICAR)处理人 ECs 可刺激 HO-1 蛋白和 mRNA 表达的浓度和时间依赖性增加,这与核因子-红细胞 2 相关因子 2(Nrf2)蛋白的明显增加有关。体内应用 AICAR 后,还观察到大鼠颈动脉中的 HO-1 诱导。AICAR 诱导的 HO-1 被 AMPK 抑制剂化合物 C、腺苷激酶抑制剂 5'-碘尿苷和 AMPK-α(1/2)沉默所阻断,并被 AMPK 激活剂 A-769662 模拟,以及通过感染表达组成型激活 AMPK-α(1)的腺病毒来感染 ECs。AICAR 还诱导 HO-1 启动子活性的显著升高,该升高被 HO-1 启动子的抗氧化反应元件突变或显性负 Nrf2 的过表达所消除。最后,AMPK 的激活抑制细胞因子介导的 EC 死亡,而这可被 HO 抑制剂锡原卟啉-IX 或沉默 HO-1 表达所预防。总之,AMPK 通过 Nrf2/抗氧化反应元件信号通路刺激人 ECs 中的 HO-1 基因表达。HO-1 的诱导介导了 AMPK 的抗细胞凋亡作用,这可能为在代谢应激期间维持 EC 活力提供了一种重要的适应性反应。