Kong Zhiwei, Pan Haichang, Wang Zi, Abla Alida, Wei Yingming
Guangxi Key Laboratory of Animal Breeding, Disease Control and Prevention, Department of Animal Sciences, Guangxi University, Nanning 530004, China.
Institute for Agricultural and Animal Husbandry Industry Development, Guangxi University, Nanning 530004, China.
Animals (Basel). 2024 Oct 18;14(20):3016. doi: 10.3390/ani14203016.
Hypoxia in the mammary gland epithelial cells of milk buffalo (BMECs) can affect milk yield and composition, and it can even cause metabolic diseases. Nitidine chloride (NC) is a natural alkaloid with antioxidant properties that can scavenge excessive reactive oxygen species (ROS). However, the effect of NC on the hypoxic injury of BMECs and its molecular mechanisms are still unknown. Here, an immunofluorescence assay, transmission electron microscopy (TEM), and flow cytometry, combined with untargeted metabolomics, were used to investigate the protective effect of NC on hypoxic stress injury in BMECs. It was found that NC can significantly reduce cell activity ( < 0.05) and inhibit cellular oxidative stress ( < 0.05) and cell apoptosis ( < 0.05). A significant decrease in mitophagy mediated by the PINK1-Parkin pathway was observed after NC pretreatment ( < 0.05). In addition, a metabolic pathway enrichment analysis demonstrated that the mechanisms of NC against hypoxic stress may be related to the downregulation of pathways involving aminoacyl tRNA biosynthesis; arginine and proline metabolism; glycine, serine, and threonine metabolism; phenylalanine, tyrosine, and tryptophan biosynthesis; and phenylalanine metabolism. Thus, NC has a protective effect on hypoxic mitochondria, and it can regulate amino acid metabolism in response to hypoxic stress. The present study provides a reference for the application of nitidine chloride to regulate the mammary lactation function of milk buffalo.
奶水牛乳腺上皮细胞（BMECs）中的缺氧会影响产奶量和乳汁成分，甚至会引发代谢性疾病。氯化两面针碱（NC）是一种具有抗氧化特性的天然生物碱，能够清除过量的活性氧（ROS）。然而，NC对BMECs缺氧损伤的影响及其分子机制仍不清楚。在此，采用免疫荧光测定法、透射电子显微镜（TEM）、流式细胞术，并结合非靶向代谢组学，研究NC对BMECs缺氧应激损伤的保护作用。结果发现，NC可显著降低细胞活性（<0.05），抑制细胞氧化应激（<0.05）和细胞凋亡（<0.05）。NC预处理后，观察到由PINK1-Parkin途径介导的线粒体自噬显著减少（<0.05）。此外，代谢途径富集分析表明，NC对抗缺氧应激的机制可能与下调涉及氨酰基tRNA生物合成、精氨酸和脯氨酸代谢、甘氨酸、丝氨酸和苏氨酸代谢、苯丙氨酸、酪氨酸和色氨酸生物合成以及苯丙氨酸代谢的途径有关。因此，NC对缺氧的线粒体具有保护作用，并且可以调节氨基酸代谢以应对缺氧应激。本研究为氯化两面针碱在调节奶水牛乳腺泌乳功能方面的应用提供了参考。
