The effects of fibroblast growth factor-21 knockdown and over-expression on its signaling pathway and glucose-lipid metabolism in vitro
The effects of fibroblast growth factor-21 knockdown and over-expression on its signaling pathway and glucose-lipid metabolism in vitro
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成纤维细胞生长因子21敲低和过表达对其信号通路和体外糖脂代谢的影响
DOI：
10.1016/j.mce.2011.07.026
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发表时间：
2012-01-02
影响因子：
4.1
通讯作者：
Yang, Gangyi
中科院分区：
文献类型：
--
作者：
Li, Ke;Li, Ling;Yang, Gangyi
目的：探讨成纤维细胞生长因子-21高表达和低表达对肝细胞和脂肪细胞糖脂代谢的影响。方法：将成纤维细胞生长因子-21高表达载体和短发夹状RNA表达载体分别导入肝癌细胞和3T3-L1脂肪细胞。用酶联免疫吸附试验检测细胞培养上清液中成纤维细胞生长因子-21的水平。Western印迹分析检测成纤维细胞生长因子-21蛋白水平，2-脱氧-D-葡萄糖摄取测定葡萄糖摄取率(Cur)。结果：在肝6肝细胞和3T3-L1脂肪细胞中，表达均显著增加(分别为4.8和4.2倍，p<0.05)，而pGenesil-fgf-21表达分别显著降低86%和78%(p<0.05)。在3T3-L1脂肪细胞中，FGFR、GLUT-1、IRS-1、ATGL、HSL和AP2mRNA的表达显著上调(p<；0.05)。除IRS-1外，FGF21基因敲除的脂肪细胞发生了相反的变化。在肝细胞中，成纤维细胞生长因子-21上调降低了HMGR和PEPCK的mRNA表达，增加了β-klotho、FGFR4和LDLR的表达(p<；0.05)。而下调则相反。结论：这些数据表明，下调和过度表达的成纤维细胞生长因子21都导致了血脂水平的变化。Cur和转录因子参与糖脂代谢，但其在脂肪细胞和肝细胞中的作用机制可能不同。(C)2011爱思唯尔爱尔兰有限公司。保留所有权利。
Objective: The aim of this study was to examine the effects of FGF-21 over- and underexpression on glucose and lipid metabolism in hepatocytes and adipocytes.Methods: FGF-21 over-expressive vectors (pcDNA-FGF-21) and FGF-21 shRNA-expressing vectors (pGenesil-FGF-21) were transfected into Hepa1-6 hepatocyte and 3T3-L1 adipocyte. The levels of FGF-21 in the incubation medium were measured by ELISA. FGF-21 protein levels by Western blot analysis, and glucose uptake rates (CUR) by measuring 2-deoxy-D-glucose uptake. The mRNA expression of transcription factors were determined by real-time quantitative PCR.Results: Transfection of pcDNA-FGF-21 significantly increased FGF-21 expression in both Hepal-6 hepatocytes and 3T3-L1 adipocytes (4.8- and 4.2-fold, respectively p < 0.05), while transfection of pGenesil-FGF-21 significantly decreased FGF-21 expressions by 86% and 78%, respectively (p < 0.05). In 3T3-L1 adipocytes, the up-regulation of FGF-21 markedly increased CUR, decreased intracellular triglyceride (TG) content, up-regulated beta-klotho, FGFR1, GLUT-1, IRS-1, ATGL, HSL, and ap2 mRNA expressions (p < 0.05). Opposite changes occurred in FGF-21 knockdown adipocytes, except for IRS-1. In hepatocyte, FGF-21 up-regulation reduced HMGR and PEPCK mRNA expression and increased beta-klotho, FGFR4 and LDLr expression (p < 0.05). whereas down-regulation had the opposite effects.Conclusion: These data suggest that both FGF-21 knockdown and over-expression led to changes in lipid levels. CUR and transcription factors involved in glucose and lipid metabolism, but that the mechanisms of FGF-21 actions in adipocytes and hepatocytes might be different. (C) 2011 Elsevier Ireland Ltd. All rights reserved.