Xu Jiajun, Peng Zhaoyun, Li Runping, Dou Tonghai, Xu Weigang, Gu Guojun, Liu Yun, Kang Zhimin, Tao Hengyi, Zhang John H, Ostrowski Robert P, Lu Jian, Sun Xuejun
Department of Diving Medicine, Second Military Medical University, Shanghai 200433, People's Republic of China.
Neurosci Lett. 2009 Feb 27;451(3):274-8. doi: 10.1016/j.neulet.2009.01.008. Epub 2009 Jan 9.
Acetazolamide has been recognized as an effective treatment for acute mountain sickness. The efficacy of acetazolamide is related to metabolic acidosis, which promotes chemoreceptors to respond to hypoxic stimuli at altitude. In this study, adult male Sprague-Dawley rats were treated with acetazolamide (100mg/kg or 50mg/kg, I.P.) for 3 days. Primary cultured cortical neurons and PC12 cell lines were exposed to acidosis-permissive (pH 6.5) or standard (pH 7.2) media for 20h. HIF-1alpha and its target genes were assayed by Western blot, real-time PCR, HIF-1 DNA-binding assay and chloramphenicol acetyltransferase reporter gene assay. HIF-1alpha protein level and HIF-1 DNA-binding activities were increased in cerebral cortices of rats treated with acetazolamide. Moreover, the mRNA levels of erythropoietin, vascular endothelial growth factor, and glucose transporter-1 also increased. The HIF-1alpha protein level and activity of HIF-driven chloramphenicol acetyltransferase reporters of cortical neurons and PC12 cells treated with acidosis media were significantly enhanced. We conclude that the normoxic induction of HIF-1alpha and HIF-1 mediated genes by acetazolamide may mediate the effect of acetazolamide in the reduction of symptoms of acute mountain sickness.
乙酰唑胺已被公认为是治疗急性高原病的有效药物。乙酰唑胺的疗效与代谢性酸中毒有关，代谢性酸中毒可促进化学感受器对高原低氧刺激作出反应。在本研究中，成年雄性Sprague-Dawley大鼠接受乙酰唑胺（100mg/kg或50mg/kg，腹腔注射）治疗3天。将原代培养的皮质神经元和PC12细胞系置于允许酸中毒的（pH 6.5）或标准的（pH 7.2）培养基中20小时。通过蛋白质免疫印迹法、实时聚合酶链反应、HIF-1 DNA结合试验和氯霉素乙酰转移酶报告基因试验检测HIF-1α及其靶基因。接受乙酰唑胺治疗的大鼠大脑皮质中HIF-1α蛋白水平和HIF-1 DNA结合活性增加。此外，促红细胞生成素、血管内皮生长因子和葡萄糖转运蛋白-1的mRNA水平也升高。用酸中毒培养基处理的皮质神经元和PC12细胞的HIF-1α蛋白水平和HIF驱动的氯霉素乙酰转移酶报告基因的活性显著增强。我们得出结论，乙酰唑胺对HIF-1α和HIF-1介导基因的常氧诱导可能介导了乙酰唑胺减轻急性高原病症状的作用。
