Wei S, Zan L S, Wang H B, Cheng G, Du M, Jiang Z, Hausman G J, McFarland D C, Dodson M V
College of Animal Science and Technology, Northwest A&F University, Yangling, Shaanxi, P.R. China.
Genet Mol Res. 2013 Feb 27;12(1):494-505. doi: 10.4238/2013.January.4.21.
Fatty acid binding protein 4 (FABP4) is an important adipocyte gene, with roles in fatty acid transport and fat deposition in animals as well as human metabolic syndrome. However, little is known about the functional regulation of FABP4 at the cellular level in bovine. We designed and selected an effective shRNA (small hairpin RNA) against bovine FABP4, constructed a corresponding adenovirus (AD-FABP4), and then detected its influence on mRNA expression of four differentiation-related genes (PPAR(y), CEBPA, CEBPB, and SREBF1) and three lipid metabolism-related genes (ADIPOQ, LEP and LEPR) of adipocytes. The FABP4 mRNA content, derived from bovine adipocytes, decreased by 41% (P < 0.01) after 24 h and 66% (P < 0.01) after 72 h of AD-FABP4 infection. However, lower mRNA content of FABP4 did not significantly alter levels of differentiation-related gene expression at 24 h following AD-FABP4 treatment of bovine-derived preadipocytes (P = 0.54, 0.78, 0.89, and 0.94, respectively). Meanwhile, knocking down (partially silencing) FABP4 significantly decreased ADIPOQ (P < 0.05) and LEP (P < 0.01) gene expression after 24 h of AD-FABP4 treatment, decreased ADIPOQ (P < 0.01) and LEP (P < 0.01) gene expression, but increased LEPR mRNA expression (P < 0.01) after a 72-h treatment of bovine preadipocytes. We conclude that FABP4 plays a role in fat deposition and metabolic syndrome by regulating lipid metabolism-related genes (such as ADIPOQ, LEP and LEPR), without affecting the ability of preadipocytes to differentiate into adipocytes.
脂肪酸结合蛋白4（FABP4）是一种重要的脂肪细胞基因，在动物的脂肪酸运输和脂肪沉积以及人类代谢综合征中发挥作用。然而，在细胞水平上，关于牛FABP4的功能调控知之甚少。我们设计并筛选了一种针对牛FABP4的有效短发夹RNA（shRNA），构建了相应的腺病毒（AD-FABP4），然后检测其对脂肪细胞中四个分化相关基因（PPAR(y)、CEBPA、CEBPB和SREBF1）以及三个脂质代谢相关基因（ADIPOQ、LEP和LEPR）mRNA表达的影响。AD-FABP4感染24小时后，源自牛脂肪细胞的FABP4 mRNA含量下降了41%（P<0.01），72小时后下降了66%（P<0.01）。然而，在AD-FABP4处理牛源前脂肪细胞24小时后，较低的FABP4 mRNA含量并未显著改变分化相关基因的表达水平（分别为P = 0.54、0.78、0.89和0.94）。同时，在AD-FABP4处理牛前脂肪细胞24小时后，敲低（部分沉默）FABP4显著降低了ADIPOQ（P<0.05）和LEP（P<0.01）基因的表达，在处理72小时后，降低了ADIPOQ（P<0.01）和LEP（P<0.01）基因的表达，但增加了LEPR mRNA的表达（P<0.01）。我们得出结论，FABP4通过调节脂质代谢相关基因（如ADIPOQ、LEP和LEPR）在脂肪沉积和代谢综合征中发挥作用，而不影响前脂肪细胞分化为脂肪细胞的能力。
