College of Animal Science and Veterinary Medicine, Shandong Agricultural University, Tai'an 271018, People's Republic of China.
Mol Biol Rep. 2011 Mar;38(3):1975-81. doi: 10.1007/s11033-010-0319-4. Epub 2010 Sep 16.
Experiments were conducted to investigate the effect of betaine supplementation on mRNA expression levels of lipogenesis genes and CpG methylation of lipoprotein lipase gene (LPL) in broilers. From 22 days of age, 78 broilers were feed basal diet without betaine and basal diet supplemented with 0.1% betaine, respectively, and at 56 and 66 days of age, the traits of 15 chickens (7 males and 8 females) of each group were recorded and abdominal fat pads were collected. The mRNA expression levels of several lipogenesis gene were analyzed by semi-quantitative RT-PCR and real-time quantitative RT-PCR (qPCR), respectively. The CpG methylation profile at the promoter region of LPL gene in 66-day-old broilers was determined by bisulfite sequencing. The average daily gain and percent abdominal fat traits were slightly improved in 56-day-old and 66-day-old broilers after dietary supplementation of betaine to diet. After adding 0.1% betaine to diet, the mRNA levels of fatty acid synthase (FAS) and adipocyte-type fatty acid-binding protein genes in abdominal adipose were significantly decreased in 56-day-old broilers, and those of LPL and FAS genes in abdominal adipose were significantly decreased in 66-day-old broilers comparing with the control group (P < 0.05 and P < 0.001). Moreover, in 66-day-old broilers fed 0.1% betaine diet, a different CpG methylation pattern was observed: the CpG dinucleotides of 1st, 6th, 7th, 8th and from 10th to 50th were less methylated; however, those of 2nd, 5th and 9th were more heavily methylated. The results suggest that transcription of some lipogenesis genes was decreased by betaine supplementation and betaine may decrease LPL mRNA expression by altering CpG methylation pattern on LPL promoter region.
试验旨在研究甜菜碱对肉鸡脂肪生成基因 mRNA 表达水平和脂蛋白脂肪酶(LPL)基因 CpG 甲基化的影响。从 22 日龄开始,78 只肉鸡分别饲喂基础日粮和基础日粮+0.1%甜菜碱,56 日龄和 66 日龄时,记录每组 15 只鸡(7 只公鸡和 8 只母鸡)的性状,采集腹部脂肪垫。采用半定量 RT-PCR 和实时定量 RT-PCR(qPCR)分别分析几种脂肪生成基因的 mRNA 表达水平。采用亚硫酸氢盐测序法测定 66 日龄肉鸡 LPL 基因启动子区 CpG 甲基化谱。日粮中添加甜菜碱可略微提高 56 日龄和 66 日龄肉鸡的平均日增重和腹部脂肪百分率。日粮中添加 0.1%甜菜碱可使 56 日龄肉鸡腹部脂肪中脂肪酸合酶(FAS)和脂肪细胞型脂肪酸结合蛋白基因的 mRNA 水平显著降低,66 日龄肉鸡腹部脂肪中 LPL 和 FAS 基因的 mRNA 水平也显著降低(P<0.05 和 P<0.001)。此外,在饲喂 0.1%甜菜碱日粮的 66 日龄肉鸡中,观察到不同的 CpG 甲基化模式:第 1、6、7、8 位和第 10 位至第 50 位的 CpG 二核苷酸甲基化程度降低;然而,第 2、5 和 9 位的 CpG 二核苷酸甲基化程度增加。结果表明,甜菜碱的添加降低了一些脂肪生成基因的转录水平,甜菜碱可能通过改变 LPL 启动子区的 CpG 甲基化模式来降低 LPL mRNA 的表达。