Zhang Ruiyang, Zhu Weiyun, Mao Shengyong
Laboratory of Gastrointestinal Microbiology, College of Animal Science and Technology, Nanjing Agricultural University, Nanjing, 210095 Peoples Republic of China.
J Anim Sci Biotechnol. 2016 Jul 29;7:42. doi: 10.1186/s40104-016-0100-1. eCollection 2016.
The objective of this study was to characterize the mRNA expression profile related to rumen epithelial inflammation through the in vivo and in vitro experiments. In the in vivo experiment, rumen papillae were collected from four dairy cows adapted to either a 40 % (LC) or 70 % (HC) concentrate feeds for microarray analysis.
Results showed that 245 differentially expressed genes (DEGs) were detected in the cows fed the HC relative to the LC diet. The DEGs were first annotated, and results revealed that the expression of inflammation-related genes, including IL-1β, IL-2, IL-22, CCL19, CCL8, CX3CR1, CXCL6, INHBE, LEPR, PRL, and TNFRSF9 found in the cytokine-cytokine receptor pathway were up-regulated in the HC-fed cows, indicating local inflammation in the rumen epithelium was triggered. The expression of IL-1β, IL-2, and IL-6 was further validated by qRT-PCR. To demonstrate whether there were relationships between cytokine mRNA expression and ruminal factors (pH and LPS), the isolated ruminal epithelial cells were cultured in vitro. Results showed that the mRNA expression of IL-1β, IL-2, IL-6, and IL-8 increased after the LPS treatment, while low-pH treatment elevated the mRNA expression of TNF-α, suggesting that low-pH coupled with higher levels of LPS in rumen of cows fed the HC may be mainly responsible for the triggered local ruminal inflammation.
Our results indicate that ruminal local inflammation response might be triggered during HC feeding, and these findings also enhance the knowledge of rumen epithelial adaptation to HC at the molecular level.
本研究的目的是通过体内和体外实验来表征与瘤胃上皮炎症相关的mRNA表达谱。在体内实验中,从四头适应40%(低浓缩料组,LC)或70%(高浓缩料组,HC)浓缩饲料的奶牛采集瘤胃乳头进行微阵列分析。
结果显示,与LC日粮组相比,HC日粮组奶牛中检测到245个差异表达基因(DEG)。首先对这些DEG进行注释,结果表明,在细胞因子-细胞因子受体途径中发现的炎症相关基因,包括IL-1β、IL-2、IL-22、CCL19、CCL8、CX3CR1、CXCL6、INHBE、LEPR、PRL和TNFRSF9在HC日粮组奶牛中的表达上调,表明瘤胃上皮局部炎症被触发。通过qRT-PCR进一步验证了IL-1β、IL-2和IL-6的表达。为了证明细胞因子mRNA表达与瘤胃因子(pH值和LPS)之间是否存在关系,体外培养分离的瘤胃上皮细胞。结果显示,LPS处理后IL-1β、IL-2、IL-6和IL-8的mRNA表达增加,而低pH处理提高了TNF-α的mRNA表达,表明HC日粮组奶牛瘤胃中低pH值与较高水平的LPS可能是触发局部瘤胃炎症的主要原因。
我们的结果表明,在HC日粮喂养期间可能会触发瘤胃局部炎症反应,这些发现也在分子水平上增强了对瘤胃上皮对HC适应的认识。