Cullin 3 mitigates nonesterified fatty acid–induced oxidative stress in mammary epithelial cells: Involvement of BCL2/BECN1 and autophagy

• DOI: 10.3168/jds.2024-25879
• Publication Date: 2025-03-06T00:06:22Z
• Authors: Yifei Dong; Wenwen Gao; Kai Liu; Cai Lin; Muhammad Usman; Juan J. Loor; Guojin Li; Liguang Cao; Zifeng Yang; Jianchun Luo; Depeng Li; Qianwen Sun; Lin Lei; Xiliang Du; Yuxiang Song; Guowen Liu; Xinwei Li
• Keywords: NEFA; GCLM

High nonesterified fatty acid (NEFA) concentrations in cows with clinical ketosis lead to metabolic dysfunction mammary cells, resulting oxidative stress. Studies have shown that autophagy is impaired the glands of ketotic cows, while enhancing mitigate stress these animals. Cullin3 (CUL3), an E3 ubiquitin ligase, integral for maintaining cellular homeostasis, particularly regulation and autophagy. Whether CUL3 involved mitigating NEFA-induced unknown. This study aimed investigate protective effects underlying mechanisms whereby mitigates epithelial cells. First, gland tissue blood samples were collected from healthy [n = 12, β-hydroxybutyrate (BHB) < 0.6 mM] (n BHB > 3.0 mM). Compared had reduced productive performance, decreased expression, autophagic activity, increased status tissue. In vitro, incubating MAC-T 1.2 mM NEFA downregulated autophagy, Adenovirus-mediated overexpression attenuated accumulation peroxides reactive oxygen species (ROS), silencing via siRNA exacerbated effects. Even when nuclear factor erythroid 2 related (NFE2L2) expression was by CUL3, there no worsening reductions mRNA levels NFE2L2 downstream target genes [NADPH quinone oxidoreductase 1 (NQO1), Heme oxygenase-1 (HMOX1), Glutamate-cysteine ligase catalytic (GCLC) Glutamate cysteine modifier subunit (GCLM)]. The reduction diminished upon 5 (ATG5) suggesting alleviates Additionally, aggravated decrease BCL2 apoptosis regulator (BCL2) alleviating beclin1(BECN1) expression. Under treatment, partly mitigated CUL3-induced elevation BECN1. Overall, characterize ketosis. activation, likely through BCL2-BECN1 pathway, enhances Thus, targeting CUL3-mediated could be a promising therapeutic strategy reduce stress-induced damage bovine