Fang Cindy X, Dong Feng, Thomas D Paul, Ma Heng, He Leilei, Ren Jun
Center for Cardiovascular Research and Alternative Medicine, University of Wyoming College of Health Sciences, Laramie, WY 82071, USA.
Am J Physiol Heart Circ Physiol. 2008 Sep;295(3):H1206-H1215. doi: 10.1152/ajpheart.00319.2008. Epub 2008 Jul 18.
Cellular hypertrophy is regulated by coordinated pro- and antigrowth machineries. Foxo transcription factors initiate an atrophy-related gene program to counter hypertrophic growth. This study was designed to evaluate the role of Akt, the forkhead transcription factor Foxo3a, and atrophy genes muscle-specific RING finger (MuRF)-1 and atrogin-1 in cardiac hypertrophy and contractile dysfunction associated with high-fat diet-induced obesity. Mice were fed a low- or high-fat diet for 6 mo along with a food-restricted high-fat weight control group. Echocardiography revealed decreased fractional shortening and increased end-systolic diameter and cardiac hypertrophy in high-fat obese but not in weight control mice. Cardiomyocytes from high-fat obese but not from weight control mice displayed contractile and intracellular Ca2+ defects including depressed maximal velocity of shortening/relengthening, prolonged duration of shortening/relengthening, and reduced intracellular Ca2+ rise and clearance. Caspase activities were greater in high-fat obese but not in weight control mouse hearts. Western blot analysis revealed enhanced basal Akt and Foxo3a phosphorylation and reduced insulin-stimulated phosphorylation of Akt and Foxo3a without changes in total protein expression of Akt and Foxo3a in high-fat obese hearts. RT-PCR and immunoblotting results displayed reduced levels of the atrogens atrogin-1 and MuRF-1, the upregulated hypertrophic markers GATA4 and ciliary neurotrophic factor receptor-alpha, as well as the unchanged calcineurin and proteasome ubiquitin in high-fat obese mouse hearts. Transfection of H9C2 myoblast cells with dominant-negative Foxo3a adenovirus mimicked palmitic acid (0.8 mM for 24 h)-induced GATA4 upregulation without an additive effect. Dominant-negative Foxo3a-induced upregulation of pAkt and repression of phosphatase and tensin homologue were abrogated by palmitic acid. These results suggest a cardiac hypertrophic response in high-fat diet-associated obesity at least in part through inactivation of Foxo3a by the Akt pathway.
细胞肥大受促生长和抗生长机制的协同调控。Foxo转录因子启动与萎缩相关的基因程序以对抗肥大生长。本研究旨在评估Akt、叉头转录因子Foxo3a以及萎缩基因肌肉特异性E3泛素连接酶1(MuRF-1)和肌肉萎缩相关基因1(atrogin-1)在高脂饮食诱导的肥胖相关心脏肥大和收缩功能障碍中的作用。将小鼠分为低脂饮食组、高脂饮食组以及饮食限制的高脂体重对照组,喂养6个月。超声心动图显示,高脂肥胖小鼠出现缩短分数降低、收缩末期直径增加和心脏肥大,而体重对照组小鼠未出现这些情况。高脂肥胖小鼠的心肌细胞表现出收缩和细胞内钙离子缺陷,包括最大缩短/舒张速度降低、缩短/舒张持续时间延长以及细胞内钙离子升高和清除减少。高脂肥胖小鼠心脏中的半胱天冬酶活性更高,但体重对照组小鼠心脏中未出现这种情况。蛋白质免疫印迹分析显示,高脂肥胖心脏中基础Akt和Foxo3a磷酸化增强,胰岛素刺激的Akt和Foxo3a磷酸化降低,而Akt和Foxo3a的总蛋白表达没有变化。逆转录聚合酶链反应(RT-PCR)和免疫印迹结果显示,高脂肥胖小鼠心脏中萎缩基因atrogin-1和MuRF-1水平降低,肥大标志物GATA4和睫状神经营养因子受体α上调,而钙调神经磷酸酶和蛋白酶体泛素水平未改变。用显性负性Foxo3a腺病毒转染H9C2成肌细胞可模拟棕榈酸(0.8 mM,处理24小时)诱导的GATA4上调,且无累加效应。棕榈酸可消除显性负性Foxo3a诱导的pAkt上调和磷酸酶及张力蛋白同源物的抑制。这些结果表明,高脂饮食相关肥胖中的心脏肥大反应至少部分是通过Akt途径使Foxo3a失活介导的。