Hagman Derek K, Hays Lori B, Parazzoli Susan D, Poitout Vincent
Pacific Northwest Research Institute, Seattle, Washington 98122, USA.
J Biol Chem. 2005 Sep 16;280(37):32413-8. doi: 10.1074/jbc.M506000200. Epub 2005 Jun 8.
Abnormalities in lipid metabolism have been proposed as contributing factors to both defective insulin secretion from the pancreatic beta cell and peripheral insulin resistance in type 2 diabetes. Previously, we have shown that prolonged exposure of isolated rat islets of Langerhans to excessive fatty acid levels impairs insulin gene transcription. This study was designed to assess whether palmitate alters the expression and binding activity of the key regulatory factors pancreas-duodenum homeobox-1 (PDX-1), MafA, and Beta2, which respectively bind to the A3, C1, and E1 elements in the proximal region of the insulin promoter. Nuclear extracts of isolated rat islets cultured with 0.5 mm palmitate exhibited reduced binding activity to the A3 and C1 elements but not the E1 element. Palmitate did not affect the overall expression of PDX-1 but reduced its nuclear localization. In contrast, palmitate blocked the stimulation of MafA mRNA and protein expression by glucose. Combined adenovirus-mediated overexpression of PDX-1 and MafA in islets completely prevented the inhibition of insulin gene expression by palmitate. These results demonstrate that prolonged exposure of islets to palmitate inhibits insulin gene transcription by impairing nuclear localization of PDX-1 and cellular expression of MafA.
脂质代谢异常被认为是2型糖尿病中胰腺β细胞胰岛素分泌缺陷和外周胰岛素抵抗的促成因素。此前,我们已经表明,将分离的大鼠胰岛长期暴露于过量脂肪酸水平会损害胰岛素基因转录。本研究旨在评估棕榈酸酯是否会改变关键调节因子胰腺十二指肠同源盒-1(PDX-1)、MafA和Beta2的表达及结合活性,这些因子分别与胰岛素启动子近端区域的A3、C1和E1元件结合。用0.5 mM棕榈酸酯培养的分离大鼠胰岛的核提取物对A3和C1元件的结合活性降低,但对E1元件的结合活性未降低。棕榈酸酯不影响PDX-1的整体表达,但会降低其核定位。相反,棕榈酸酯阻断了葡萄糖对MafA mRNA和蛋白质表达的刺激作用。胰岛中联合腺病毒介导的PDX-1和MafA过表达完全阻止了棕榈酸酯对胰岛素基因表达的抑制。这些结果表明,胰岛长期暴露于棕榈酸酯会通过损害PDX-1的核定位和MafA的细胞表达来抑制胰岛素基因转录。