Mitochondrial oxidative damage in chicken skeletal muscle induced by acute heat stress
Mitochondrial oxidative damage in chicken skeletal muscle induced by acute heat stress
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DOI：
10.2141/jpsa.44.439
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发表时间：
2007-10-01
影响因子：
1.5
通讯作者：
Toyomizu, Masaaki
中科院分区：
文献类型：
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作者：
Mujahid, Ahmad;Pumford, Neil R.;Toyomizu, Masaaki
长期以来，线粒体在生理条件下维持生物能量状态中起着关键作用。我们以前的研究表明，急性热应激增加了鸡骨骼肌线粒体活性氧（ROS）的产生。这种增加的ROS产生可能导致脂质和蛋白质的非特异性修饰，然后可能导致生物能量功能障碍。如果热应激鸡的线粒体ROS产生增加导致氧化损伤，则可以检测到线粒体过氧化脂质和氧化修饰蛋白的变化。为了研究这一点，将3周龄的雄性肉鸡（n = 4-8）暴露于急性热应激（34 ℃持续18小时），而对照鸡保持在热中性条件（25 ℃）。分离骨骼肌肌膜下线粒体，并用于研究线粒体丙二醛（MDA）和蛋白质羰基。用0.01%丁基羟基甲苯（BHT）比色法测定线粒体硫代巴比妥酸反应物质（TBARS）的形成，并表示为MDA当量。为了检测氧化敏感的线粒体蛋白，通过二维（2D）电泳解析的多肽用DNP特异性抗体进行免疫染色，使用蛋白质印迹法检测羰基化蛋白。在热应激鸡，线粒体MDA是2.7倍高，82个线粒体蛋白质被氧化时，与对照组相比。这些结果表明，在热应激鸡，增加线粒体ROS的生产导致线粒体脂质和蛋白质的氧化损伤。
Mitochondria have long been known to play a critical role in maintaining bioenergetic status under physiological conditions. Our previous studies have shown that acute heat stress increases production of mitochondrial reactive oxygen species (ROS) in the skeletal muscle of chickens. This increased ROS production may lead to nonspecific modification of lipids and proteins, which may then result in bioenergetic dysfunctions. If increased mitochondrial ROS production in heat-stressed chickens causes oxidative damage, changes in mitochondrial peroxidized lipids and oxidatively modified proteins can be detected. To study this, 3-week-old male broiler chickens (n = 4-8) were exposed to acute heat stress (34 degrees C for 18 h) while control chickens were kept at thermoneutral condition (25 degrees C). Skeletal muscle subsarcolemmal mitochondria were isolated and used to study mitochondrial malondialdehyde (MDA) and protein carbonyl groups. Mitochondrial thiobarbituric acid reactive substances (TBARS) formation was measured colorimetrically with 0.01% butylated hydroxytoluene (BHT) and expressed as MDA equivalent. To detect oxidation-sensitive mitochondrial proteins, polypeptides resolved by two-dimensional (2D) electrophoresis were immunostained with DNP-specific antibodies for carbonylated proteins using Western blotting. In heat-stressed chickens, mitochondrial MDA was 2.7-fold higher, and 82 mitochondrial proteins were oxidized when compared to that of control chickens. These results suggest that in heat-stressed chickens, increased mitochondrial ROS production leads to oxidative damage to mitochondrial lipids and proteins.