Yi Bao, Wang Jigui, Wang Shuang, Yuan Daoli, Sun Jiazeng, Li Zhili, Mao Yaping, Hou Qiang, Liu Weiquan
Department of Biochemistry and Molecular Biology, State Key Laboratory of Agrobiotechnology, College of Biological Sciences, China Agricultural University, No. 2 Yuanmingyuan West Road, Haidian District, Beijing, 100193, China.
Cell Biol Int. 2014 Aug;38(8):918-23. doi: 10.1002/cbin.10285. Epub 2014 Apr 28.
Fatty acid binding protein 3 (H-FABP, FABP3) has been significantly associated with intramuscular fat (IMF) content in pigs, which is positively correlated with palatability of pork. However, its underlying function is not fully elucidated. We have investigated the effects of overexpression of the FABP3 gene on differentiation and adipogenesis of 3T3-L1 preadipocytes in the fat Banna mini-pig inbred line (fBMIL). Eukaryotic vectors that expressed the FABP3 protein were constructed, and stably established in the 3T3-L1 preadipocytes cell line. Cells were induced in a standard differentiation cocktail. Morphological changes and the degree of adipogenesis were measured by Oil Red O staining assay and triacylglycerol content measurement, respectively. mRNA expression levels of triacylglycerol metabolism-related genes were measured by qPCR. FABP3 significantly promoted differentiation of 3T3-L1 cells and enhanced triacylglycerol levels (P < 0.05). mRNA of the peroxisome proliferator-activated receptor γ (PPARγ), adipocyte fatty acid binding protein (422/aP2) and glycerol-3-phosphate dehydrogenase (GPDH) gene increased markedly (P < 0.05). In conclusion, expression of the FABP3 gene enhances adipogenesis in 3T3-L1 preadipocytes primarily by upregulating lipogenic PPARγ, 422/aP2 and GPDH genes.
脂肪酸结合蛋白3(H-FABP,FABP3)与猪的肌内脂肪(IMF)含量显著相关,而肌内脂肪含量与猪肉的口感呈正相关。然而,其潜在功能尚未完全阐明。我们研究了FABP3基因过表达对版纳微型猪近交系脂肪型(fBMIL)3T3-L1前脂肪细胞分化和脂肪生成的影响。构建了表达FABP3蛋白的真核载体,并在3T3-L1前脂肪细胞系中稳定建立。细胞在标准分化混合液中诱导。分别通过油红O染色法和三酰甘油含量测定法测量形态变化和脂肪生成程度。通过qPCR测量三酰甘油代谢相关基因的mRNA表达水平。FABP3显著促进3T3-L1细胞的分化并提高三酰甘油水平(P<0.05)。过氧化物酶体增殖物激活受体γ(PPARγ)、脂肪细胞脂肪酸结合蛋白(422/aP2)和甘油-3-磷酸脱氢酶(GPDH)基因的mRNA显著增加(P<0.05)。总之,FABP3基因的表达主要通过上调生脂PPARγ、422/aP2和GPDH基因来增强3T3-L1前脂肪细胞的脂肪生成。