[目的]研究禁抗后饲粮中添加罗伊氏乳杆菌LR1对仔猪各肠段微小RNA(microRNA,miRNA)表达的影响,为罗伊氏乳杆菌LR1调控仔猪肠道功能的机制研究筛选潜在miRNA.[方法]选取21日龄体重相近的杜×长×大断奶仔猪144头,随机分为3组,每组8个重复,每个重复6头仔猪.对照组仔猪饲喂基础饲粮,抗生素组和益生菌组仔猪在饲喂基础饲粮的基础上分别添加复合抗生素(100 mg/kg喹乙醇+75 mg/kg金霉素)及5×1010 CFU罗伊氏乳杆菌LR1,试验期42 d.试验结束后每个重复随机屠宰1头仔猪,采集空肠组织进行miRNA高通量测序分析,采用实时荧光定量PCR检测筛选miRNA在仔猪十二指肠、空肠、回肠和结肠中的表达量.[结果]饲粮中添加罗伊氏乳杆菌LR1可引起仔猪肠道组织miRNA表达发生明显变化,改变了空肠高表达miRNA的排序.差异表达miRNA的潜在靶基因主要富集在代谢相关通路及PI3K-Akt、MAPK、Chemokine、Ras信号通路.实时荧光定量PCR显示,3个组间仔猪空肠差异miRNA的表达结果与转录组测序结果大致相同,其中,表达上调的miRNA在对照组和抗生素组间表达趋势较为一致,表达下调的miRNA在对照组和益生菌组间表达趋势较为一致,而肠道高表达miRNA在3个组中均有较为一致的表达趋势.[结论]禁抗后饲粮中添加罗伊氏乳杆菌LR1会引起肠道miRNA表达发生变化,筛选出的差异表达miRNA可为罗伊氏乳杆菌LR1调控仔猪肠道发育及营养吸收代谢等研究提供基础.
[Objective]This study was conducted to investigate the effects of Lactobacillus reuteri LR1 replacing antibiotics on the expression of miRNA in different intestinal segments of piglets,so as to screen potential miRNA for studying the mechanism of Lactobacillus reuteri LR1 on regulating intestinal function of piglets.[Method]A total of 144 cross-bred(DurocX LandraceX Yorkshire)weaned piglets with similar body weight at 21 days old were selected and randomly divided into 3 groups,with 8 replicates in each group and 6 piglets in each replicate.The piglets in control group were fed with the basic diet,the piglets in antibiotic and probiotic groups were fed with the basic diet supplemented with compound antibiotics(100 mg/kg quinoline ethanol+75 mg/kg aureomycin)and 5X1010 CFU Lactobacillus reuteri LR1,respectively.The experiment lasted for 42 days.At the end of the experiment,1 piglet from each replicate was randomly selected and slaughtered,the jejunum tissue was collected for miRNA high-throughput sequencing and analysis.The expression of the selected miRNA in duodenum,jejunum,ileum and colon were detected by Real-time quantitative PCR.[Result]Lactobacillus reuteri LR1 supplementation caused significant changes in miRNA expression,altering the sort of highly expressed miRNA in jejunum.The potential target genes of differentially expressed miRNA were mainly enriched in metabolic related pathways,PI3K-Akt,MAPK,Chemokine and Ras signaling pathways.Real-time quantitative PCR results showed that the expression of differential miRNA in jejunum among 3 groups had the same result with RNA-Seq.Moreover,the expression of up-regulated miRNA showed a relatively consistent trend between control and antibiotic groups,down-regulated miRNA showed a relatively consistent trend between control and probiotic groups,while the highly expressed miRNA in intestine showed a relatively consistent expression trend among 3 groups.[Conclusion]Adding Lactobacillus reuteri LR1 in diet replacing antibiotics could cause changes in the expression of intestinal miRNA.The differentially expressed miRNA screened could provide a foundation for research on the regulating intestinal development and nutrient absorption metabolism by Lactobacillus reuteri LR1 in piglets.
李虹仪;杨雪芬;林泽森;吴晓菲;吕祺欣;余佳莉;张茂
韶关学院生物与农业学院,韶关 512000广东省农业科学院动物科学研究所,猪禽种业全国重点实验室,农业农村部华南动物营养与饲料重点实验室,广东省畜禽育种与营养研究重点实验室,岭南现代农业科学与技术广东省实验室茂名分中心,广州 510640
畜牧业
罗伊氏乳杆菌;仔猪;肠道miRNA
Lactobacillus reuteri;piglets;intestinal miRNA
《中国畜牧兽医》 2024 (001)
74-85 / 12
广东省基础与应用基础研究(2021A1515010483);韶关市支持科研工作者项目(210715124531392、210725204531009)
10.16431/j.cnki.1671-7236.2024.01.008
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