Key Laboratory of Ruminant Molecular and Cellular Breeding of Ningxia Hui Autonomous Region, College of Animal Science and Technology, Ningxia University, Yinchuan, China.
Ningxia Borui Ruminant Nutrition Research Center Co., Ltd, Yinchuan, China.
Anim Biotechnol. 2024 Nov;35(1):2381080. doi: 10.1080/10495398.2024.2381080. Epub 2024 Aug 1.
Mastitis, a serious threat to the health and milk production function of dairy cows decreases milk quality. Blood from three healthy cows and three mastitis cows were collected in this study and their transcriptome was sequenced using the Illumina HiSeq platform. Differentially expressed genes (DEGs) were screened according to the |log2FoldChange| > 1 and -value < 0.05 criteria. Pathway enrichment and functional annotation were performed through KEGG and GO analyses. Finally, the mechanism of the AMP-activated protein kinase (AMPK) mediation of (-)-epigallocatechin-3-gallate (EGCG) to promote lipid metabolism in mastitis cows was analyzed in bovine mammary epithelial cells (BMECs). Transcriptome analysis revealed a total of 825 DEGs, with 474 genes showing increased expression and 351 genes showing decreased expression. The KEGG analysis of DEGs revealed that they were mainly linked to tumour necrosis factor, nuclear factor-κB signalling pathway, and lipid metabolism-related signalling pathway, whereas GO functional annotation found that DEGs were enriched in threonine and methionine kinase activity, cellular metabolic processes, and cytoplasm. AMPK expression, which is involved in several lipid metabolism pathways, was downregulated in mastitis cows. The results of in vitro experiments showed that the inhibition of AMPK promoted the expression of lipid synthesis genes in lipopolysaccharide-induced BMECs and that EGCG could promote lipid synthesis by decreasing the expression of AMPK and downregulating the expression of inflammatory factors in inflammatory BMECs. In conclusion, our study demonstrated that AMPK mediated EGCG to inhabit of inflammatory responses and promote of lipid synthesis in inflammatory BMECs.
乳腺炎是奶牛健康和产奶功能的严重威胁，会降低牛奶质量。本研究采集了 3 头健康奶牛和 3 头乳腺炎奶牛的血液，使用 Illumina HiSeq 平台进行转录组测序。根据 |log2FoldChange| > 1 和 -value < 0.05 的标准筛选差异表达基因 (DEGs)。通过 KEGG 和 GO 分析进行通路富集和功能注释。最后，在牛乳腺上皮细胞 (BMEC) 中分析了 AMP 激活蛋白激酶 (AMPK) 介导 (-)-表没食子儿茶素没食子酸酯 (EGCG) 促进乳腺炎奶牛脂代谢的机制。转录组分析共发现 825 个 DEGs，其中 474 个基因表达上调，351 个基因表达下调。DEGs 的 KEGG 分析表明，它们主要与肿瘤坏死因子、核因子-κB 信号通路和脂代谢相关信号通路有关，而 GO 功能注释发现 DEGs 富集于苏氨酸和蛋氨酸激酶活性、细胞代谢过程和细胞质。参与多种脂代谢途径的 AMPK 表达在乳腺炎奶牛中下调。体外实验结果表明，AMPK 的抑制促进了脂多糖诱导的 BMECs 中脂质合成基因的表达，而 EGCG 可以通过降低炎症 BMECs 中 AMPK 的表达和下调炎症因子的表达来促进脂质合成。总之，本研究表明 AMPK 介导 EGCG 抑制炎症反应并促进炎症 BMECs 中的脂质合成。
