RNA-Seq data of ruminal epithelial tissue and 16S rRNA sequencing data of rumen digesta in goats infusion of three short-chain fatty acids [RNA-seq]

Emerging data has highlighted the importance of short-chain fatty acids (SCFAs), particularly butyrate, in regulating ruminal homeostasis in vivo isolated epithelial cells. However, little is known about other SCFAs like acetate or propionate, and the interaction between rumen microbes and epithelial immunity are rarely reported. Here, we firstly combined infusion of three SCFAs, to study their different roles in ruminal development, antioxidant capacity, barrier functions, and immunity, as well as cross-talk with ruminal microbiome (16S rRNA sequencing data of rumen digesta) and derived transcriptome (RNA-Seq) and metabolism using an in vivo goat model. A total of twenty-four healthy multiparous Guanzhong goats with an average body weight of 47.44 ± 3.38 kg at 1.5 years old in latter half of gestation period were selected in this study. All dairy goats were then randomly divided into four groups (n = 6 for each group) after adaption with the diet and light conditions to receive different SCFAs infusion treatments. They were assigned to a normal control group infusion with normal saline (NC), an infusion with sodium acetate solution (SA), an infusion with sodium propionate solution (SP), and an infusion with sodium butyrate solution (SB) according to previous studies in ruminants. The same volume of SCFAs and normal saline solutions at 1 L were slowly perfused into the rumen with a rumen tube daily before morning feeding. All solutions were calibrated to be consistent with a pH meter before infusion. The experiment lasted for 13 days, which combined with a 12-days of infusion period and a 1-day of sampling period. All goats were fasted for 12 hours before the sampling day. The animals were then weighed, and slaughtered and dressed by professional butchers. Firstly, the rumen digesta was immediately collected and stored directly in liquid nitrogen for the analysis of rumen microbiome using 16S rRNA sequencing. Then, another part of 5 g rumen epithelial tissue was rapidly frozen in liquid nitrogen for rumen epithelial RNA-Seq.

近年来的新兴研究数据表明，短链脂肪酸（short-chain fatty acids, SCFAs），尤其是丁酸，在调控体内分离的瘤胃上皮细胞稳态方面具有重要意义。然而，目前对于乙酸、丙酸等其他短链脂肪酸的研究仍较为匮乏，瘤胃微生物与上皮免疫之间的互作机制也鲜有报道。本研究首次联合输注三种短链脂肪酸，利用山羊体内模型，探究其在瘤胃发育、抗氧化能力、屏障功能与免疫调控中的不同作用，以及与瘤胃微生物组（瘤胃食糜16S rRNA测序数据）、转录组（RNA-Seq）和代谢组的互作关系。本研究共选取24只健康的经产关中奶山羊，实验时处于妊娠中后期，年龄为1.5岁，平均体重为47.44±3.38 kg。所有奶山羊在适应日粮与光照条件后，被随机分为4组（每组n=6），分别接受不同的短链脂肪酸输注处理。根据反刍动物既往研究，将各组分别设为：生理盐水输注正常对照组（normal saline, NC）、乙酸钠溶液输注组（sodium acetate, SA）、丙酸钠溶液输注组（sodium propionate, SP）以及丁酸钠溶液输注组（sodium butyrate, SB）。每日于晨间饲喂前，通过瘤胃插管将1 L等体积的短链脂肪酸溶液或生理盐水缓慢灌注至瘤胃内。输注前，所有溶液均通过pH计校准至一致的pH值。本实验共计13天，其中包含12天的输注周期与1天的采样周期。采样前12小时，所有山羊禁食。随后对山羊进行称重，由专业屠宰人员进行屠宰与胴体处理。首先，立即采集瘤胃食糜并直接置于液氮中保存，用于后续16S rRNA测序分析瘤胃微生物组。此外，另取5 g瘤胃上皮组织快速置于液氮中冷冻，用于瘤胃上皮的RNA-Seq分析。