Targeted suppression of calpain-10 expression impairs insulin-stimulated glucose uptake in cultured primary human skeletal muscle cells
Targeted suppression of calpain-10 expression impairs insulin-stimulated glucose uptake in cultured primary human skeletal muscle cells
复制标题
DOI：
10.1016/j.ymgme.2007.05.001
复制
发表时间：
2007-08-01
影响因子：
3.8
通讯作者：
Walker, Mark
中科院分区：
文献类型：
--
作者：
Brown, Audrey E.;Yeaman, Stephen J.;Walker, Mark
Calpain-10被认为是一种新的2型糖尿病易感基因，但其增加2型糖尿病易感性的机制尚不清楚。由于骨骼肌是2型糖尿病中葡萄糖处置的外周胰岛素抵抗的主要部位，我们研究了在培养的人骨骼肌细胞中靶向抑制钙蛋白酶-10表达是否直接影响胰岛素作用。使用短干扰RNA（siRNA）来特异性抑制CAPN 10基因表达。通过定量PCR和蛋白质印迹法评估，在转录和蛋白质水平均观察到抑制。CAPN 10 mRNA表达抑制（75%。与未转染的肌管（1.43 +/-0.16-倍增加）相比，胰岛素刺激的葡萄糖摄取显著减少（p = 0.04）（相对于基础增加1.03 +/-0.06 [平均SEM]-倍）。与此相反，钙蛋白酶-10表达的抑制减少不影响胰岛素刺激的糖原合成，也不影响胰岛素刺激的蛋白激酶13的磷酸化，胰岛素信号通路的关键组成部分，这项研究证实，钙蛋白酶-10在胰岛素刺激的葡萄糖摄取在人类骨骼肌细胞中发挥作用。钙蛋白酶-10表达的抑制不影响胰岛素刺激的糖原合成，也不影响胰岛素信号通过PKB，这表明钙蛋白酶-10可能发挥直接的调节作用后，葡萄糖摄取机制。(c)2007年爱思唯尔公司All rights reserved.
Calpain-10 was identified as a novel type 2 diabetes susceptibility gene, although the mechanisms by which it increases susceptibility to type 2 diabetes remain unclear. As skeletal muscle is the principal site of the peripheral insulin resistance for glucose disposal in type 2 diabetes, we investigated whether targeted suppression of calpain-10 expression directly affects insulin action in cultured human skeletal muscle cells. Short interfering RNAs (siRNAs) were employed to specifically suppress CAPN10 gene expression. Suppression was seen at both the transcript and protein level, as assessed by quantitative PCR and Western blotting. Suppression of CAPN10 mRNA expression (75%. decrease compared to untransfected myotubes) was associated with a significant decrease (p = 0.04) in insulin-stimulated glucose uptake (1.03 +/- 0.06 [mean SEM]-fold increase over basal) compared to the untransfected myotubes (1.43 +/- 0.16-fold increase). In contrast, decreased suppression of calpain-10 expression did not affect insulin-stimulated glycogen synthesis nor insulin-stimulated phosphorylation of protein kinase 13, a key component of the insulin-signalling pathway.This study confirms that calpain-10 plays a role in insulin-stimulated glucose uptake in human skeletal muscle cells. Suppression of calpain-10 expression did not affect insulin-stimulated glycogen synthesis nor insulin-signalling via PKB, suggesting that calpain-10 may exert a direct regulatory effect upon the glucose uptake mechanism. (c) 2007 Elsevier Inc. All rights reserved.