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Abstract：
BACKGROUND: Folic acid is a water-soluble B vitamin (B9), which is closely related to the body\'s immune and other metabolic pathways. The folic acid synthesized by rumen microbes has been unable to meet the needs of high-yielding dairy cows. The incidence rate of subclinical mastitis in dairy herds worldwide ranged between 25%~65% with no obvious symptoms, but it significantly causes a decrease in lactation and milk quality. Therefore, this study aims at exploring the effects of folic acid supplementation on the expression profile of lncRNAs, exploring the molecular mechanism by which lncRNAs regulate immunity in subclinical mastitic dairy cows.
RESULTS: The analysis identified a total of 4384 lncRNA transcripts. Subsequently, differentially expressed lncRNAs in the comparison of two groups (SF vs. SC, HF vs. HC) were identified to be 84 and 55 respectively. Furthermore, the weighted gene co-expression network analysis (WGCNA) and the KEGG enrichment analysis result showed that folic acid supplementation affects inflammation and immune response-related pathways. The two groups have few pathways in common. One important lncRNA MSTRG.11108.1 and its target genes (ICAM1, CCL3, CCL4, etc.) were involved in immune-related pathways. Finally, through integrated analysis of lncRNAs with GWAS data and animal QTL database, we found that differential lncRNA and its target genes could be significantly enriched in SNPs and QTLs related to somatic cell count (SCC) and mastitis, such as MSTRG.11108.1 and its target gene ICAM1, CXCL3, GRO1.
CONCLUSIONS: For subclinical mastitic cows, folic acid supplementation can significantly affect the expression of immune-related pathway genes such as ICAM1 by regulating lncRNAs MSTRG.11108.1, thereby affecting related immune phenotypes. Our findings laid a ground foundation for theoretical and practical application for feeding folic acid supplementation in subclinical mastitic cows.
RESULTS: The analysis identified a total of 4384 lncRNA transcripts. Subsequently, differentially expressed lncRNAs in the comparison of two groups (SF vs. SC, HF vs. HC) were identified to be 84 and 55 respectively. Furthermore, the weighted gene co-expression network analysis (WGCNA) and the KEGG enrichment analysis result showed that folic acid supplementation affects inflammation and immune response-related pathways. The two groups have few pathways in common. One important lncRNA MSTRG.11108.1 and its target genes (ICAM1, CCL3, CCL4, etc.) were involved in immune-related pathways. Finally, through integrated analysis of lncRNAs with GWAS data and animal QTL database, we found that differential lncRNA and its target genes could be significantly enriched in SNPs and QTLs related to somatic cell count (SCC) and mastitis, such as MSTRG.11108.1 and its target gene ICAM1, CXCL3, GRO1.
CONCLUSIONS: For subclinical mastitic cows, folic acid supplementation can significantly affect the expression of immune-related pathway genes such as ICAM1 by regulating lncRNAs MSTRG.11108.1, thereby affecting related immune phenotypes. Our findings laid a ground foundation for theoretical and practical application for feeding folic acid supplementation in subclinical mastitic cows.
摘要：
背景：叶酸是一种水溶性B族维生素（B9），它与人体的免疫和其他代谢途径密切相关。瘤胃微生物合成的叶酸已不能满足高产奶牛的需要。全球奶牛群亚临床乳腺炎的发病率在25%~65%之间,无明显症状,但它显著导致泌乳和牛奶质量下降。因此,本研究旨在探索补充叶酸对lncRNAs表达谱的影响，探索lncRNAs调节亚临床奶牛免疫的分子机制。
结果:分析鉴定了总共4384个lncRNA转录物。随后，两组比较中差异表达的lncRNAs(SFvs.SC,HFvs.HC)分别鉴定为84和55。此外，加权基因共表达网络分析(WGCNA)和KEGG富集分析结果显示,补充叶酸影响炎症和免疫应答相关通路。两组有几个共同的途径。lncRNA重要有MSTRG.11108.1及其靶基因(ICAM1、CCL3、CCL4等。)参与免疫相关途径。最后,通过与GWAS数据和动物QTL数据库整合分析lncRNAs，我们发现差异lncRNA及其靶基因可以在与体细胞计数（SCC）和乳腺炎相关的SNP和QTLs中显著富集，如MSTRG.11108.1及其靶基因ICAM1、CXCL3、GRO1。
结论：对于亚临床乳牛，补充叶酸可以通过调节lncRNAsMSTRG.11108.1显著影响免疫相关通路基因如ICAM1的表达,从而影响相关免疫表型。我们的研究结果为亚临床奶牛饲喂叶酸的理论和实践应用奠定了基础。
结果:分析鉴定了总共4384个lncRNA转录物。随后，两组比较中差异表达的lncRNAs(SFvs.SC,HFvs.HC)分别鉴定为84和55。此外，加权基因共表达网络分析(WGCNA)和KEGG富集分析结果显示,补充叶酸影响炎症和免疫应答相关通路。两组有几个共同的途径。lncRNA重要有MSTRG.11108.1及其靶基因(ICAM1、CCL3、CCL4等。)参与免疫相关途径。最后,通过与GWAS数据和动物QTL数据库整合分析lncRNAs，我们发现差异lncRNA及其靶基因可以在与体细胞计数（SCC）和乳腺炎相关的SNP和QTLs中显著富集，如MSTRG.11108.1及其靶基因ICAM1、CXCL3、GRO1。
结论：对于亚临床乳牛，补充叶酸可以通过调节lncRNAsMSTRG.11108.1显著影响免疫相关通路基因如ICAM1的表达,从而影响相关免疫表型。我们的研究结果为亚临床奶牛饲喂叶酸的理论和实践应用奠定了基础。
