棕榈酸对模拟高原缺氧大鼠离体脑线粒体解耦联蛋白活性和质子漏的影响.pdfVIP

生理学报 Acta Physiologica Sinica, February 25, 2008, 60 (1): 59-64 59 研究论文 棕榈酸对模拟高原低氧大鼠离体脑线粒体解耦联蛋白活性及 质子漏的影响 * 徐 瑜，柳君泽 ，夏 琛 第三军医大学病理生理学教研室，重庆 400038 摘 要：本文旨在通过观察棕榈酸对模拟高原低氧大鼠离体脑线粒体解耦联蛋白(uncoupling proteins, UCPs)活性的影响及脑 线粒体质子漏与膜电位的改变，探讨UCPs 在介导游离脂肪酸对低氧时线粒体氧化磷酸化功能改变中的作用。将Sprague- Dawley 大鼠随机分为对照组、急性低氧组和慢性低氧组。低氧大鼠于低压舱内模拟海拔5 000 m 高原23 h/d 作低氧暴露， 分别连续低氧3 d 和30 d 。用差速密度梯度离心法提取脑线粒体，[3H]-GTP 法测定UCPs 含量与活性，TPMP+ 电极与Clark 氧电极结合法测量线粒体质子漏，罗丹明123 荧光法测定线粒体膜电位。结果显示，低氧使脑线粒体内UCPs 含量与活性 升高、质子漏增加、线粒体膜电位降低；同时，低氧暴露降低脑线粒体对棕榈酸的反应性，UCPs 活性的改变率低于对 照组，且线粒体UCPs 含量、质子漏、膜电位变化率亦出现相同趋势。线粒体质子漏与反映UCPs 活性的Kd 值呈线性负 相关(P0.01, r = -0.906) ，与反映UCPs 含量的Bmax 呈线性正相关(P0.01, r = 0.856) ，与膜电位呈线性负相关(P0.01, r = -0.880) 。以上结果提示，低氧导致的脑线粒体质子漏增加及膜电位降低与线粒体内UCPs 活性升高有关，同时低氧暴 露能降低脑线粒体对棕榈酸的反应性，提示在高原低氧环境下，游离脂肪酸升高在维持线粒体能量代谢中起着自身保护和 调节机制。 关键词：棕榈酸；线粒体解耦联蛋白；低氧；脑；高原病 中图分类号：R 3 39 . 5 ；Q 4 9 4 Effects of palmitic acid on activity of uncoupling proteins and proton leak in in vitro cerebral mitochondria from the rats exposed to simulated high altitude hypoxia * XU Yu, LIU Jun-Ze , XIA Chen Department of Pathophysiology and Altitude Physiology, the Third Military Medical University, Chongqing 400038, China Abstract: To reveal the roles of uncoupling proteins (UCPs) in disorder of mitochondrial oxidative phosphorylation induced by free fatty acid during hypoxic exposure, the effects of palmitic acid on activity of UCPs, proton leak and mitochondrial membrane potential in hypoxia-exposed rat brain mitochondria were observed in vitro. Adult Sprague-Dawley (SD) rats were set randomly into control, acute hypoxia and chronic hypoxia groups (n=8 in each group). The acute and chronic hypoxic rats were exposed to simulated 5 000 m high altitude in a hypobaric chamber 23 h/d for 3 d and 30 d, respectively. The brain mitochondria were isolated by centrifugation. UCP conten