AMPK磷酸化障碍导致烧伤后骨骼肌脂质沉积的-第三军医大学学报.DOC

DOI：10.16016/j.1000-5404.201412252 腺苷酸激活蛋白激酶磷酸化障碍导致烧伤后骨骼肌脂质沉积的实验研究 王 超，吴 丹，王宏宇，胡建红，张 勇，彭 曦（400038 重庆，第三军医大学西南医院全军烧伤研究所，创伤、烧伤与复合伤国家重点实验室） [摘要] 目的 研究腺苷酸激活蛋白激酶（AMP activated protein kinase, AMPK）/乙酰辅酶a羧化酶(Acetyl CoA carboxylase, ACC）信号通路对烧伤后骨骼肌脂质沉积(intramyocellular lipids, IMCLs)的影响并探讨其机制。 方法 采用30％体表面积Ⅲ度烧伤小鼠模型，将54只BALB/C小鼠随机分为正常对照组、AMPK激活剂(acadesine，AICAR)组、烧伤组、烧伤+AICAR组，同位素标记法检测各组小鼠腓肠肌中肉碱脂酰转移酶-1(carnitine palmitoyl transferase-1, CPT1)活性变化，Western blot检测各组小鼠腓肠肌中AMPK-α、p-AMPK-α、ACC及p-ACC表达水平，通过油红O染色和甘油三酯(triglyceride，TG)测定法评估各组小鼠腓肠肌IMCLs情况。 结果 烧伤后小鼠腓肠肌中ACC表达量较伤前显著升高(P0.05)，而AMPK-α、p-AMPK-α、p-ACC以及CPT1活性均显著降低(P0.05)。在AICAR作用下，正常小鼠腓肠肌中p-AMPK-α表达显著升高[(1.16±0.08)vs(2.38±0.22)，P0.05],p-ACC表达显著升高[(1.74±0.10)vs(3.72±0.18)，P0.05]，CPT1活性显著升高[(2.95±0.39)vs(6.35±0.68)，P0.05]，TG含量显著降低[(3.88±0.40)vs(2.89±0.54)，P0.05]。烧伤+AICAR组小鼠腓肠肌中p-AMPK-α、p-ACC、CPT1活性及甘油三酯含量较烧伤组无显著差异(P0.05)。正常对照组和AICAR组正常小鼠腓肠肌组织切片油红O染色未见明显脂质沉积，而烧伤组和烧伤+AICAR组烧伤小鼠腓肠肌组织切片油红O染色均显示有大量脂质沉积，两组间并无显著差异(P0.05)。结论 骨骼肌中AMPK磷酸化障碍是烧伤后IMCLs的重要分子机制之一。 [关键词]烧伤；骨骼肌脂质沉积；腺苷酸激活蛋白激酶；乙酰辅酶a羧化酶；肉碱脂酰转移酶-1 [中图法分类号] R644 ［文献标志码］ A [基金项目] 国家自然科学基金面上项目 [通信作者] 彭 曦，E-mail: pxlrmm@163.com AMPK phosphorylation disorder induces intramyocellular lipids deposition in mice after burn injury Wang Chao, Wu Dan, Wang Hongyu, Hu Jianhong, Zhang Yong, Peng Xi (State Key Laboratory of Trauma, Burns and Combined Injury, Institute of Burns, Southwest Hospital, Third Military Medical University, Chongqing, 400038, China) [Abstract]Objective To determine the effect of AMP activated protein kinase (AMPK)/acetyl CoA carboxylase (ACC) signal pathway on the deposition of intramyocellular lipids (IMCLs) and investigate its underlying mechanisms. Methods Fifty-four BALB/C mice were inflicted with 30% total body surface area full-thickness burns, and then randomly divided into 4 groups, that is, control, acadesine treatment (AICAR, AMPK activator), burn control, and burn-AICAR treatment groups. First, mitochondria were isolated to tes