Saeedi Ramesh, Saran Varun V, Wu Sherry S Y, Kume Erika S, Paulson Kim, Chan Annie P K, Parsons Hannah L, Wambolt Richard B, Dyck Jason R B, Brownsey Roger W, Allard Michael F
James Hogg iCAPTURE Centre for Cardiovascular and Pulmonary Research, Department of Pathology and Laboratory Medicine, University of British Columbia-St. Paul's Hospital, Vancouver, British Columbia, Canada V6Z 1Y6.
Am J Physiol Heart Circ Physiol. 2009 Jun;296(6):H1822-32. doi: 10.1152/ajpheart.00396.2008. Epub 2009 Apr 17.
Substrate use switches from fatty acids toward glucose in pressure overload-induced cardiac hypertrophy with an acceleration of glycolysis being characteristic. The activation of AMP-activated protein kinase (AMPK) observed in hypertrophied hearts provides one potential mechanism for the acceleration of glycolysis. Here, we directly tested the hypothesis that AMPK causes the acceleration of glycolysis in hypertrophied heart muscle cells. The H9c2 cell line, derived from the embryonic rat heart, was treated with arginine vasopressin (AVP; 1 microM) to induce a cellular model of hypertrophy. Rates of glycolysis and oxidation of glucose and palmitate were measured in nonhypertrophied and hypertrophied H9c2 cells, and the effects of inhibition of AMPK were determined. AMPK activity was inhibited by 6-[4-(2-piperidin-1- yl-ethoxy)-phenyl]-3-pyridin-4-yl-pyrrazolo-[1,5-a]pyrimidine (compound C) or by adenovirus-mediated transfer of dominant negative AMPK. Compared with nonhypertrophied cells, glycolysis was accelerated and palmitate oxidation was reduced with no significant alteration in glucose oxidation in hypertrophied cells, a metabolic profile similar to that of intact hypertrophied hearts. Inhibition of AMPK resulted in the partial reduction of glycolysis in AVP-treated hypertrophied H9c2 cells. Acute exposure of H9c2 cells to AVP also activated AMPK and accelerated glycolysis. These elevated rates of glycolysis were not altered by AMPK inhibition but were blocked by agents that interfere with Ca(2+) signaling, including extracellular EGTA, dantrolene, and 2-aminoethoxydiphenyl borate. We conclude that the acceleration of glycolysis in AVP-treated hypertrophied heart muscle cells is partially dependent on AMPK, whereas the acute glycolytic effects of AVP are AMPK independent and at least partially Ca(2+) dependent.
在压力超负荷诱导的心肌肥大中,底物利用从脂肪酸转向葡萄糖,糖酵解加速是其特征。在肥大心脏中观察到的AMP激活的蛋白激酶(AMPK)的激活为糖酵解加速提供了一种潜在机制。在此,我们直接检验了AMPK导致肥大心肌细胞中糖酵解加速这一假说。源自胚胎大鼠心脏的H9c2细胞系用精氨酸加压素(AVP;1μM)处理以诱导肥大的细胞模型。在未肥大和肥大的H9c2细胞中测量葡萄糖和棕榈酸酯的糖酵解速率及氧化速率,并确定抑制AMPK的作用。AMPK活性通过6-[4-(2-哌啶-1-基-乙氧基)-苯基]-3-吡啶-4-基-吡唑并-[1,5-a]嘧啶(化合物C)或通过腺病毒介导的显性负性AMPK的转移来抑制。与未肥大细胞相比,肥大细胞中糖酵解加速而棕榈酸酯氧化减少,葡萄糖氧化无显著改变,这种代谢特征与完整的肥大心脏相似。抑制AMPK导致AVP处理的肥大H9c2细胞中糖酵解部分降低。H9c2细胞急性暴露于AVP也激活AMPK并加速糖酵解。这些升高的糖酵解速率不受AMPK抑制的影响,但被干扰Ca(2+)信号传导的试剂所阻断,包括细胞外EGTA、丹曲林和2-氨基乙氧基二苯硼酸。我们得出结论,AVP处理的肥大心肌细胞中糖酵解加速部分依赖于AMPK,而AVP的急性糖酵解作用不依赖于AMPK且至少部分依赖于Ca(2+)。