脂多糖与钙离子共处理的牛乳腺上皮细胞miRNA表达谱的比较分析

doi:10.16431/j.cnki.1671-7236.2024.09.035

Abstract

Abstract: 【Objective】 This study was aimed to investigate the regulation of calcium ions on inflammatory responses in bovine mammary epithelial cells, and compare and analyze miRNA expression profiles of bovine mammary epithelial cells co-treated with lipopolysaccharide and calcium ions. 【Method】 A model of inflammatory response was created by treating bovine mammary epithelial cells with lipopolysaccharide.The transcript levels of inflammatory factors were measured in bovine mammary epithelial cells co-treated with different concentrations (0.5-6.6 mmol/L) of calcium ions and lipopolysaccharide (50 μg/mL).RNA was extracted from bovine mammary epithelial cell samples with normal, lipopolysaccharide treated and co-treated with lipopolysaccharide and calcium ions, with 3 samples in each group.Quality assessment was performed on 9 RNA samples, followed by miRNA sequencing and bioinformatics analysis.Finally, Real-time quantitative PCR validation was performed on common differentially expressed miRNA among 3 groups. 【Result】 In a model of inflammation in bovine mammary epithelial cells, high concentrations of calcium ions were found to inhibit the transcription of inflammatory factors.Nine RNA samples were obtained, with clean reads accounting for between 93.86% and 98.18% of raw reads.The sequence distribution lengths were all concentrated between 21 and 24 nt.The samples from all three groups exhibited high intra-group similarity but also high inter-group variability.A total of 47 miRNA were identified as differentially expressed across 3 groups of samples.The samples among 3 groups showed differential expression of bta-miR-11980, bta-miR-1246 and bta-miR-677.The results of GO function and KEGG pathway enrichment analysis showed that the target genes of differentially expressed miRNA were significantly enriched in GO entries related to regulation of cell communication, signaling regulation, endomembrane system, phylogeny, cellular developmental process and cell differentiation, etc.The genes targeted by the differentially expressed miRNA were significantly enriched in various pathways, such as the HIF-1 signaling pathway, ErbB signaling pathway, TNF signaling pathway, synaptic vesicle cycle and inositol phosphate metabolism pathway.Real-time quantitative PCR results confirmed that the expression of common differentially expressed miRNA among 3 sample groups (bta-miR-677 and bta-miR-1246) were consistent with RNA-Seq results. 【Conclusion】 Calcium ions indirectly affected the inflammatory response in bovine mammary epithelial cells by regulating the expression of differentially expressed miRNA, such as bta-miR-11980, bta-miR-1246 and bta-miR-677.

Key words: lipopolysaccharide; calcium ion; bovine mammary epithelial cells; miRNA expression profile

CLC Number:

S852.32

XU Haotian, YU Yuetong, LI Jing, MA Zhiyuan, YANG Bin, WANG Zekun, TUO Haixin, QI Meng. Comparative Analysis of miRNA Expression Profiles in Bovine Mammary Epithelial Cells Treated with Lipopolysaccharide and Calcium Ions[J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(9): 4066-4079.

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Comparative Analysis of miRNA Expression Profiles in Bovine Mammary Epithelial Cells Treated with Lipopolysaccharide and Calcium Ions

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