Effects of hypoxia stress on the milk synthesis in bovine mammary epithelial cells

Milk synthesis is an energy-intensive process influenced by oxygen availability. This study investigates how hypoxia affects milk synthesis in BMECs, focusing on key genes involved in lactation and energy metabolism. BMECs were cultured in a normoxic environment and then transferred to a hypoxia chamber with 1% O2 for specified durations. The study evaluated cellular responses through various molecular experiments and RNA sequencing. Small interfering RNA was employed to knock down HIF-1α to investigate whether the lactation-related phenotype alteration depends on HIF-1α. Hypoxia disrupted milk protein production by reducing mTOR/P70S6K/4EBP1 signaling and downregulating genes critical for amino acid transport and protein synthesis. Triglyceride synthesis increased due to enhanced fatty acid uptake and the upregulation of regulatory proteins, including FASN and PPARγ. Although glucose uptake was elevated under hypoxia, key enzymes for lactose synthesis were downregulated, suggesting a redirection of glucose toward energy production. Mitochondrial function was impaired under hypoxia, with reduced gene expression in TCA cycle, ETC, cytosol-mitochondrial transport, decreased ATP levels, increased ROS levels, and structural alterations. Additionally, lipid synthesis and glucose uptake depend on HIF-1α, while milk protein synthesis alterations occurred independently of HIF-1α. Hypoxia alters milk synthesis in BMECs by disrupting milk protein synthesis, enhancing lipid metabolism, and impairing energy production. These findings provide valuable insights into the molecular mechanisms underlying the effect of oxygen deprivation on lactation efficiency, offering potential targets for mitigating hypoxic stress in the mammary glands of dairy animals.