Department of Biotechnology, Daegu University, Kynungsan, Kyungbuk, Republic of Korea.
Proteomics. 2010 Aug;10(15):2870-81. doi: 10.1002/pmic.200900815.
In this study, the effects of capsaicin on expression of skeletal muscle proteins in Sprague-Dawley rats fed with a high-fat diet (HFD) were investigated. Rats were fed a HFD with or without capsaicin treatment for 8 wk. After HFD feeding, capsaicin-treated rats weighed an average of 8% less than those of the HFD control group. Gastrocnemius muscle tissue from lean and obese rats with or without capsaicin treatment was arrayed using 2-DE for detection of HFD-associated markers. Proteomic analysis using 2-DE demonstrated that 36 spots from a total of approximately 600 matched spots showed significantly different expression; 27 spots were identified as gastrocnemius muscle proteins that had been altered in response to capsaicin feeding, and 6 spots could not be identified by mass fingerprinting. Expression of various muscle proteins was determined by immunoblot analysis for the determination of molecular mechanisms, whereby capsaicin caused inhibition of adipogenesis. Immunoblot analysis revealed increased uncoupling protein 3 (UCP3) protein expression in HFD-fed rats, whereas contents were reduced with capsaicin treatment. Compared with the HFD control group, capsaicin treatment increased phosphorylation of AMP-activated protein kinase (AMPIC) CP3 and acetyl-CoA carboxylase (ACC). To support this result, we also analyzed in vitro differential protein expression in L6 skeletal muscle cells. These data suggest that the AMPK-ACC-malonyl-CoA metabolic signaling pathway is one of the targets of capsaicin action. To the best of our knowledge, this is the first proteomic study to report on analysis of diet-induced alterations of protein expression that are essential for energy expenditure in rat muscle.
本研究旨在探讨辣椒素对高脂肪饮食(HFD)喂养的 Sprague-Dawley 大鼠骨骼肌蛋白表达的影响。大鼠给予 HFD 喂养,并分别用或不用辣椒素处理 8 周。HFD 喂养后,辣椒素处理组大鼠的体重平均比 HFD 对照组低 8%。用 2-DE 排列瘦鼠和肥胖鼠的腓肠肌组织,以检测 HFD 相关标记物。用 2-DE 进行蛋白质组学分析表明,在总共约 600 个匹配点中,有 36 个点的表达差异显著;27 个点被鉴定为对辣椒素喂养有反应的腓肠肌蛋白,6 个点的质量指纹图谱无法识别。通过免疫印迹分析确定了各种肌肉蛋白的表达,以确定分子机制,即辣椒素抑制脂肪生成。免疫印迹分析显示,UCP3 蛋白在 HFD 喂养的大鼠中表达增加,而用辣椒素处理后含量降低。与 HFD 对照组相比,辣椒素处理增加了 AMP 激活蛋白激酶(AMPIC)CP3 和乙酰辅酶 A 羧化酶(ACC)的磷酸化。为了支持这一结果,我们还分析了 L6 骨骼肌细胞中体外差异蛋白表达。这些数据表明,AMPK-ACC-丙二酰辅酶 A 代谢信号通路是辣椒素作用的靶点之一。据我们所知,这是首次对饮食诱导的大鼠肌肉能量消耗相关蛋白表达变化进行蛋白质组学分析的研究。