Globular and full-length forms of adiponectin mediate specific changes in glucose and fatty acid uptake and metabolism in cardiomyocytes
Globular and full-length forms of adiponectin mediate specific changes in glucose and fatty acid uptake and metabolism in cardiomyocytes
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DOI：
10.1016/j.cardiores.2007.04.011
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发表时间：
2007-07-01
影响因子：
10.8
通讯作者：
Sweeney, Gary
中科院分区：
文献类型：
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作者：
Palanivel, Rengasamy;Fang, Xiangping;Sweeney, Gary
目的：我们的目的是研究脂联素的球状（gAd）和全长（fAd）形式对心肌细胞中葡萄糖和脂肪酸代谢的调节。方法：我们在哺乳动物表达系统中产生fAd（由高、中、低分子量寡聚物组成），在细菌中产生gAd。这些用于处理原代新生大鼠心肌细胞（长达 48 小时），我们采用 H-3 或 C-14 标记的底物来监测葡萄糖摄取和随后通过氧化、糖原合成或乳酸产生以及脂肪酸摄取和氧化进行的代谢。采用酶法测定乙酰辅酶A羧化酶活性，并通过免疫印迹细胞裂解物测定蛋白质磷酸化和表达。脂联素受体 (AdipoR) 亚型的作用通过 siRNA 介导的敲低来确定。结果：gAd 或 fAd 响应时，葡萄糖摄取和氧化最初（1 小时）增加。 gAd或fAd刺激脂肪酸摄取，并且通过24It观察到乙酰辅酶A羧化酶活性降低和脂肪酸氧化升高。 48小时后，脂肪酸氧化增加与葡萄糖氧化和丙酮酸脱氢酶活性降低相关，而糖原合成和乳酸产量增加。 gAd 和 fAd 均引发 AMP 激酶、胰岛素受体底物 1、Akt 和糖原合酶激酶 3 β 的磷酸化。 AdipoR1 或 AdipoR2 的敲低减弱了 gAd 和 fAd 对脂肪酸摄取和氧化的影响。只有 AdipoR1 敲低才能阻止 gAd (1 h) 增加葡萄糖摄取和氧化的能力；然而，减少 AdipoR1 或 AdipoR2 的表达会减弱 gAd 的长期（24 小时）影响。 结论：这些结果清楚地表明，gAd 和 fAd 通过 AdipoR1 和 AdipoR2 对心肌细胞能量代谢产生不同且时间依赖性的影响。 (c) 2007 年欧洲心脏病学会。由 Elsevier B.V. 出版。保留所有权利。
Objective: Our aim was to investigate the regulation of glucose and fatty acid metabolism in cardiomyocytes by the globular (gAd) and full-length (fAd) forms of adiponectin.Methods: We produced fAd (consisting of high, medium and low molecular weight oligomers) in a mammalian expression system and gAd in bacteria. These were used to treat primary neonatal rat cardiomyocytes (up to 48 h), and we employed H-3- or C-14-labeled substrates to monitor glucose uptake and subsequent metabolism via oxidation, glycogen synthesis or lactate production and fatty acid uptake and oxidation. Enzymatic assay for acetyl CoA carboxylase activity was employed, and protein phosphorylation and expression was determined by immunoblotting cell lysates. The role of adiponectin receptor (AdipoR) isoforms was determined via siRNA-mediated knockdown.Results: There was an initial (I h) increase in glucose uptake and oxidation in response to gAd or fAd. Fatty acid uptake was stimulated by gAd or fAd, and by 24 It a decrease in acetyl CoA carboxylase activity and elevated fatty acid oxidation were observed. After 48 h increased fatty acid oxidation correlated with decreased glucose oxidation and pyruvate dehydrogenase activity, while glycogen synthesis and lactate production increased. Both gAd and fAd elicited phosphorylation of AMP kinase, insulin receptor substrate-1, Akt and glycogen synthase kinase-3 beta. Knockdown of AdipoR1 or AdipoR2 attenuated the effect of both gAd and fAd on fatty acid uptake and oxidation. Only AdipoR1 knockdown prevented the ability of gAd (1 h) to increase glucose uptake and oxidation; however, reducing either AdipoR1 or AdipoR2 expression attenuated the long-term (24 h) effects of gAd.Conclusions: These results clearly demonstrate that gAd and fAd mediate distinct and time-dependent effects on cardiomyocyte energy metabolism via AdipoR1 and AdipoR2. (c) 2007 European Society of Cardiology. Published by Elsevier B.V. All rights reserved.