State Key Laboratory for Zoonotic Diseases, College of Veterinary Medicine, Jilin University, 1977 Xinzhu Road, Changchun, 130062, China.
Department of First Hospital, Jilin University, 1 Xinmin Street, Changchun, 130021, China.
BMC Vet Res. 2023 Feb 11;19(1):44. doi: 10.1186/s12917-023-03588-2.
IFN-γ is a pleiotropic cytokine that has been shown to affect multiple cellular functions of bovine mammary epithelial cells (BMECs) including impaired milk fat synthesis and induction of malignant transformation via depletion of arginine, one of host conditionally essential amino acids. But the molecular mechanisms of these IFN-γ induced phenotypes are still unknown.
BMECs were treated with IFN-γ for 6 h, 12 h, and 24 h. The metabolomic profiling in BMECs upon IFN-γ induction were assessed using untargeted ultra-performance liquid chromatography-mass spectrometry (UPLC-MS) metabolomic analysis. Key differentially expressed metabolites (DEMs) were quantified by targeted metabolomics.
IFN-γ induction resulted in significant differences in the contents of metabolites. Untargeted analysis identified 221 significantly DEMs, most of which are lipids and lipid-like molecules, organic acids and derivatives, organ heterocyclic compounds and benzenoids. According to Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis, DEMs were enriched in fructose and mannose metabolism, phosphotransferase system (PTS), β-alanine metabolism, arginine and proline metabolism, methane metabolism, phenylalanine metabolism, and glycolysis/gluconeogenesis. Quantification of selected key DEMs by targeted metabolomics showed significantly decreased levels of D-(-)-mannitol, argininosuccinate, and phenylacetylglycine (PAG), while increased levels in S-hydroxymethylglutathione (S-HMG) and 2,3-bisphospho-D-glyceric acid (2,3-BPG).
These results provide insights into the metabolic alterations in BMECs upon IFN-γ induction and indicate potential theoretical basis for clarifying IFN-γ-induced diseases in mammary gland.
IFN-γ 是一种多效细胞因子，已被证明会影响牛乳腺上皮细胞 (BMEC) 的多种细胞功能，包括通过耗尽精氨酸（宿主条件必需氨基酸之一）来抑制乳脂合成和诱导恶性转化。但是，这些 IFN-γ 诱导表型的分子机制尚不清楚。
用 IFN-γ 处理 BMEC 6 h、12 h 和 24 h。使用非靶向超高效液相色谱-质谱联用 (UPLC-MS) 代谢组学分析评估 IFN-γ 诱导后 BMEC 中的代谢组学特征。通过靶向代谢组学定量关键差异表达代谢物 (DEM)。
IFN-γ 诱导导致代谢物含量有显著差异。非靶向分析鉴定出 221 个显著 DEM，其中大多数是脂质和类脂分子、有机酸及其衍生物、有机杂环化合物和苯类化合物。根据京都基因与基因组百科全书 (KEGG) 途径分析，DEM 富集在果糖和甘露糖代谢、磷酸转移酶系统 (PTS)、β-丙氨酸代谢、精氨酸和脯氨酸代谢、甲烷代谢、苯丙氨酸代谢和糖酵解/糖异生中。通过靶向代谢组学对选定的关键 DEM 进行定量分析显示，D-(-)-甘露醇、精氨琥珀酸和苯乙酰甘氨酸 (PAG) 的水平明显降低，而 S-羟甲基谷胱甘肽 (S-HMG) 和 2,3-双磷酸-D-甘油酸 (2,3-BPG) 的水平升高。
这些结果提供了 IFN-γ 诱导 BMEC 代谢改变的见解，并为阐明乳腺中 IFN-γ 诱导的疾病提供了潜在的理论基础。
