Hypoxia Affects Mitochondrial Protein Expression in Rat Skeletal Muscle
Hypoxia Affects Mitochondrial Protein Expression in Rat Skeletal Muscle
复制标题
缺氧影响大鼠骨骼肌中线粒体蛋白的表达。
DOI：
10.1089/omi.2011.0023
复制
发表时间：
2012-03-01
影响因子：
3.3
通讯作者：
Gao, Wenxiang
中科院分区：
文献类型：
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作者：
Chen, Jian;Gao, Yuqi;Gao, Wenxiang
缺氧影响哺乳动物线粒体功能，以及基于线粒体的能量代谢。详细的机制尚未完全理解。本研究应用蛋白质组学方法检测了低压缺氧大鼠线粒体各组分蛋白质的表达水平。成年雄性Wistar大鼠随机分为低氧组（海拔4,500 m，30天）和常氧对照组（海平面）。提取并纯化腓肠肌线粒体。用Clark氧电极测定线粒体耗氧量，用罗丹明123荧光探针测定线粒体跨膜电位。利用2-DE和MALDI-TOF MS分析，我们确定了8个线粒体蛋白质点，在缺氧组与常氧对照组相比，差异表达。这些蛋白质包括F1-ATP酶A链、电压依赖性阴离子通道1（VDAC）、羟酰辅酶A脱氢酶α亚基、线粒体F1复合物γ亚基、雄激素调节蛋白和三联体基序蛋白50。其中两个点，VDAC和ATP合成酶α-亚基，通过Western印迹分析得到证实。状态3呼吸时的耗氧量、呼吸控制率（RCR）对照组显著高于低氧组;线粒体跨膜电位低氧组显著高于对照组。利用多种蛋白质组学分析技术的成功应用，证明了30 d低氧暴露对线粒体ATP合成和脂质代谢相关蛋白的表达有影响，降低了线粒体膜的稳定性，并影响了线粒体电子传递链。
Hypoxia affects mammalian mitochondrial function, as well as mitochondria-based energy metabolism. The detail mechanism has not been fully understood. In this study, we detected protein expression levels in mitochondrial fractions of Wistar rats exposed to hypobaric hypoxia by use of proteomic methods. Adult male Wistar rats were randomized into an hypoxic (4,500 m, 30 days) group and a normoxic control group (sea level). Gastrocnemius muscles mitochondria were extracted and purified. Mitochondrial oxygen consumption was measured with a Clark oxygen electrode; mitochondrial transmembrane potential was detected with Rhodamine 123 as a fluoresce probe. Using 2-DE and MALDI-TOF MS analysis, we identified eight mitochondrial protein spots that were differentially expressed in the hypoxic group compared with the normoxic control. These proteins included Chain A of F1-ATPase, voltage dependent anion channel 1 (VDAC), hydroxyacyl Coenzyme A dehydrogenase α-subunit, mitochondrial F1 complex γ-subunit, androgen-regulated protein and tripartite motif protein 50. Two of the spots, VDAC and ATP synthase α-subunit, were confirmed by Western blotting analysis. Oxygen consumption during State 3 respiration, as well as the respiratory control ratio (RCR) was significantly higher in the control than that in the hypoxic group; mitochondrial transmembrane potential was significantly higher in hypoxic group than that in the control. With successful use of multiple proteomic analysis techniques, we demonstrates that 30 days hypoxia exposure has effects on the expression of mitochondrial proteins involved in ATP production and lipid metabolism, decrease the stability of mitochondrial membrane, and affect the mitochondrial electron transport chain.