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首页> 外文期刊>Journal of Cellular Physiology >Challenging of AS160/TBC1D4 Alters Intracellular Lipid milieu in L6 Myotubes Incubated With Palmitate
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Challenging of AS160/TBC1D4 Alters Intracellular Lipid milieu in L6 Myotubes Incubated With Palmitate

机译：挑战AS160 / TBC1D4改变细胞内脂质在16种肌管孵化环境棕榈酸酯

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The Akt substrate of 160 kDa (AS160) is a key regulator of GLUT4 translocation from intracellular depots to the plasma membrane in myocytes. Likely, AS160 also controls LCFAs transport, which requires relocation of fatty acid transporters. The aim of the present study was to determine the impact of AS160 knockdown on lipid milieu in L6 myotubes incubated with palmitate (PA). Therefore, we compared two different settings, namely: 1) AS160 knockdown prior to palmitate incubation (pre-PA-silencing, AS160(-)/PA); 2) palmitate incubation with subsequent AS160 knockdown (post-PA-silencing, PA/AS160(-)). The efficiency of AS160 silencing was checked at mRNA and protein levels. The expression and localization of FA transporters were determined using Western Blot and immunofluorescence analyses. Intracellular lipid content (FFA, DAG, TAG, and PL) and FA composition were estimated by GLC, whereas basal palmitate uptake was analyzed by means of scintigraphy. Both groups with silenced AS160 were characterized by a greater expression of FA transporters (FAT/CD36, FATP-1, 4) which had contributed to an increased FA cellular influx. Accordingly, we observed that post-PA-silencing of AS160 resulted in a marked decrement in DAG, TAG, and PL contents, but increased FFA content (PA/AS160(-) vs. PA). The opposite effect was observed in the group with pre-PA-silencing of AS160 in which AS160 knockdown did not affect the lipid pools (AS160(-)/PA vs. PA). Our results indicate that post-PA-silencing of AS160 has a capacity to decrease the lipotoxic effect(s) of PA by decreasing the content of lipids (DAG and PL) that promote insulin resistance in myotubes. (C) 2016 The Authors. Journal of Cellular Physiology Published by Wiley Periodicals Inc.
机译：160 kDa的Akt衬底(AS160)是一个关键监管机构的GLUT4易位细胞内仓库的质膜细胞。运输,需要搬迁的脂肪酸转运蛋白。是确定的影响AS160击倒脂质在16种肌管孵化环境棕榈酸酯(PA)。不同的设置,即:1)AS160击倒棕榈酸酯前孵化(pre-PA-silencingAS160 (-) / PA);随后AS160击倒(post-PA-silencingPA / AS160(-))。检查在mRNA和蛋白水平。FA转运蛋白的表达和定位测定用免疫印迹和吗免疫荧光分析。内容(FFA、DAG、标签和PL)和足总成分被相关估计,而基底棕榈酸酯进行了分析通过吸收显像。特点是更多表达的FA转运蛋白(脂肪/ CD36 FATP-1 4)导致增加了FA细胞涌入。因此,我们观察到post-PA-silencingDAG AS160导致显著衰减,标签,和PL内容,但FFA含量增加(PA / AS160(-)和PA)。观察组与pre-PA-silencing的AS160 AS160击倒并没有影响脂质池(AS160 (-) / PA与PA)。表明AS160 post-PA-silencing的能力降低lipotoxic效应(年代)PA减少脂质(DAG的内容PL),促进肌管的胰岛素抵抗。作者(C) 2016。生理学由威利期刊出版公司。

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