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RNA-seq 揭示了 miR-223 在缓解奶牛乳腺上皮细胞炎症中的作用。

RNA-seq reveals the role of miR-223 in alleviating inflammation of bovine mammary epithelial cells.

机构信息

School of Agriculture, Ningxia University, Yinchuan, China; Key Laboratory of Ruminant Molecular Cell Breeding, Ningxia Hui Autonomous Region, Yinchuan, China.

School of Agriculture, Ningxia University, Yinchuan, China; Key Laboratory of Ruminant Molecular Cell Breeding, Ningxia Hui Autonomous Region, Yinchuan, China.

出版信息

Res Vet Sci. 2023 Jun;159:257-266. doi: 10.1016/j.rvsc.2023.04.012. Epub 2023 Apr 19.

Abstract

Bovine mammary epithelial cells (bMECs) are involved in the early defense against the invasion of intramammary pathogens and are essential for the health of bovine mammary gland. MicroRNA (MiRNA) is a key factor that regulates cell state and physiological function. In the present study, the transcriptome profiles of miR-223 inhibitor transfection group (miR-223_Inhibitor) and negative control inhibitor transfection group (NC_Inhibitor) within bMECs were detected via the RNA sequencing (RNA-seq) platform. Based on these experiments, the differentially expressed mRNAs (DE-mRNAs) of the miR-223_Inhibitor transfection group were screened, and the Gene Ontology and Kyoto Encyclopedia of Genes and Genomes functional analyses of DE-mRNAs were performed. The results revealed that compared with the NC_Inhibitor, 224 differentially expressed genes (DEGs) were identified in the miR-223_Inhibitor, including 184 upregulated and 40 downregulated genes. The functional annotation of the above DEGs indicated that some of these genes are involved in the immune response generated by extracellular substance stimulation, regulation of the activity of cytokines and chemokines, and the immune signaling pathways of NF-κB and TNF. Meanwhile, miR-223_inhibitor upregulated the immune key genes IRF1 and NFκBIA, cytokines IL-6 and IL-24, as well as chemokines CXCL3, CXCL5, and CCR6, triggering a signaling cascade response that exacerbated inflammation in bMECs. These results suggested that miR-223 plays an important role in inhibiting the inflammatory response and maintaining the stability of bMECs, and is a potential target for treating mastitis in dairy cows.

摘要

牛乳腺上皮细胞(bMEC)参与了对乳腺内病原体入侵的早期防御,是牛乳腺健康的关键。microRNA(miRNA)是调节细胞状态和生理功能的关键因素。本研究通过 RNA 测序(RNA-seq)平台检测了 miR-223 抑制剂转染组(miR-223_Inhibitor)和阴性对照抑制剂转染组(NC_Inhibitor)内 bMEC 中的转录组谱。基于这些实验,筛选出 miR-223_Inhibitor 转染组的差异表达 mRNA(DE-mRNA),并对 DE-mRNA 进行了基因本体论和京都基因与基因组百科全书功能分析。结果表明,与 NC_Inhibitor 相比,miR-223_Inhibitor 中转录水平上调的基因有 184 个,下调的基因有 40 个,共鉴定出 224 个差异表达基因(DEGs)。上述 DEGs 的功能注释表明,其中一些基因参与了细胞外物质刺激产生的免疫反应、细胞因子和趋化因子活性的调节以及 NF-κB 和 TNF 的免疫信号通路。同时,miR-223_inhibitor 上调了免疫关键基因 IRF1 和 NFκBIA、细胞因子 IL-6 和 IL-24 以及趋化因子 CXCL3、CXCL5 和 CCR6,触发了信号级联反应,加剧了 bMEC 中的炎症。这些结果表明,miR-223 在抑制炎症反应和维持 bMEC 稳定性方面发挥着重要作用,是治疗奶牛乳腺炎的潜在靶点。

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