Wang Jing, Yin Xuemei, Sun Li, Sun Shouyong, Zi Chen, Zhu Guoqiang, Wu Shenglong, Bao Wenbin
Key Laboratory for Animal Genetics, Breeding, Reproduction and Molecular Design of Jiangsu Province, College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China.
College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China.
Int J Mol Sci. 2014 Jun 18;15(6):10989-98. doi: 10.3390/ijms150610989.
Diarrhea and edematous disease are two major causes of mortality in postweaning piglets, and these conditions lead to huge economic losses in the swine industry. E. coli F18 is the primary causative agent of these two diseases. Bactericidal/permeability-increasing protein (BPI) plays an important role in the natural defense of the host. The aim of this study was to determine the correlation between BPI gene upstream CpG island methylation and mRNA expression. In this study, bisulfite sequencing PCR (BSP) was used to detect the methylation status of the BPI gene upstream CpG island and fluorescence quantitative PCR was used to detect BPI expression in the duodenum of piglets from birth to weaning age. BPI upstream CpG islands were shown to have many putative transcription factor binding sites, 10 CpG sites and every CpG site was methylated. The CpG island methylation level was lowest in 30-day piglets and was significantly lower than levels in 8-day piglets (p<0.05). BPI mRNA expression was significantly higher in 30-day piglets than at any other age (p<0.05). Pearson's correlation analysis showed that the methylation status of the CpG island was negatively correlated with BPI mRNA expression. Statistical significances were found in CpG_1, CpG_3, CpG_4, CpG_7 and CpG_10 (p<0.05). The data indicate that BPI expression is improved by demethylation of the BPI gene upstream CpG island. Furthermore, CpG_1, CpG_3, CpG_4, CpG_7 and CpG_10 may be critical sites in the regulation of BPI gene expression.
腹泻和水肿病是断奶仔猪死亡的两大主要原因，这些病症给养猪业造成了巨大的经济损失。大肠杆菌F18是这两种疾病的主要病原体。杀菌/通透性增加蛋白（BPI）在宿主的天然防御中发挥着重要作用。本研究的目的是确定BPI基因上游CpG岛甲基化与mRNA表达之间的相关性。在本研究中，采用亚硫酸氢盐测序PCR（BSP）检测BPI基因上游CpG岛的甲基化状态，并采用荧光定量PCR检测出生至断奶期仔猪十二指肠中BPI的表达。结果显示，BPI上游CpG岛有许多假定的转录因子结合位点、10个CpG位点，且每个CpG位点均发生甲基化。30日龄仔猪的CpG岛甲基化水平最低，且显著低于8日龄仔猪（p<0.05）。30日龄仔猪的BPI mRNA表达显著高于其他任何年龄段（p<0.05）。Pearson相关性分析表明，CpG岛的甲基化状态与BPI mRNA表达呈负相关。在CpG_1、CpG_3、CpG_4、CpG_7和CpG_10位点发现有统计学意义（p<0.05）。数据表明，BPI基因上游CpG岛去甲基化可提高BPI表达。此外，CpG_1、CpG_3、CpG_4、CpG_7和CpG_10可能是BPI基因表达调控的关键位点。
