Key Laboratory of Animal Physiology and Biochemistry, Nanjing Agricultural University, Nanjing 210095, People's Republic of China.
Br J Nutr. 2013 May 28;109(10):1731-8. doi: 10.1017/S0007114512003728. Epub 2012 Sep 10.
To investigate whether the effect of maternal dietary protein on offspring lipid metabolism is mediated by microRNA (miRNA), fourteen Meishan sows were fed either low-protein (LP, half of standard protein (SP) level, n 7) or SP (n 7) diets throughout gestation and lactation periods. PPAR-γ and CCAAT/enhancer-binding protein-β (C/EBP-β) protein expression was evaluated. The expression of miRNA predicted to directly target PPAR-γ and C/EBP-β in the subcutaneous fat of offspring at weaning age was determined, and the functions of these potential miRNA were verified. The results showed that piglet body weight and back fat thickness were significantly decreased in the LP group compared with the SP group (P<0·05). The protein level of PPAR-γ was significantly decreased and C/EBP-β protein expression was also decreased, though not significantly (P=0·056), in the subcutaneous fat of the LP group. Furthermore, miRNA expression analysis showed that miR-130b, targeting the PPAR-γ 3'-untranslated region (UTR), and miR-374b, targeting the C/EBP-β 3'-UTR, were significantly increased in the LP group compared with the SP group; other candidate regulatory miRNA were expressed similarly in both groups. Dual luciferase activity assay results indicated that miR-130b directly recognised and bound to the 3'-UTR of PPAR-γ and thereby suppressed PPAR-γ gene expression. Similar results were found for miR-374b and the 3'-UTR of C/EBP-β. The present study showed that miR-130b and miR-374b are involved in the effect of maternal dietary protein on offspring lipid metabolism in pigs. These results shed new light on our understanding of the maternal effect on offspring lipid deposition.
为了研究母体膳食蛋白质对后代脂质代谢的影响是否通过 microRNA（miRNA）介导，14 头梅山母猪在妊娠和哺乳期分别喂食低蛋白（LP，标准蛋白（SP）水平的一半，n=7）或 SP（n=7）饮食。评估 PPAR-γ 和 CCAAT/增强子结合蛋白-β（C/EBP-β）蛋白表达。测定断奶后代皮下脂肪中预测可直接靶向 PPAR-γ 和 C/EBP-β 的 miRNA 的表达，并验证这些潜在 miRNA 的功能。结果表明，与 SP 组相比，LP 组仔猪体重和背脂厚度显著降低（P<0.05）。LP 组皮下脂肪中 PPAR-γ 蛋白水平显著降低，C/EBP-β 蛋白表达也降低，尽管差异不显著（P=0.056）。此外，miRNA 表达分析显示，miR-130b 靶向 PPAR-γ 3'-UTR，miR-374b 靶向 C/EBP-β 3'-UTR，在 LP 组中显著高于 SP 组；两组中其他候选调节 miRNA 的表达相似。双荧光素酶活性测定结果表明，miR-130b 可直接识别并结合 PPAR-γ 的 3'-UTR，从而抑制 PPAR-γ 基因表达。miR-374b 与 C/EBP-β 的 3'-UTR 也有类似的结果。本研究表明，miR-130b 和 miR-374b 参与了母体膳食蛋白质对后代脂质代谢的影响。这些结果为我们理解母体对后代脂质沉积的影响提供了新的视角。
