AMPK-endoplasmic reticulum stress axis contributes to lipopolysaccharide-caused mitochondrial dysfunction by regulating mitochondria-associated membrane function in bovine hepatocytes

• Published in: Journal of dairy science 2023-07, Vol.106 (7), p.5146-5164
• Main Authors: Xie, Wan, Chen, Mengru, Loor, Juan J., Song, Xiaokun, Ma, Nana, Zhou, Shendong, Zhang, Hongzhu, Chang, Guangjun, Shen, Xiangzhen
• Format: Article
• Language: English
• Subjects: adenosine 5′-monophosphate-activated protein kinase; AMP-Activated Protein Kinases - metabolism; Animals; Apoptosis; bovine hepatocytes; Cattle; Endoplasmic Reticulum Stress; Endoribonucleases - metabolism; Female; Hepatocytes - metabolism; JNK Mitogen-Activated Protein Kinases - metabolism; Lipopolysaccharides - metabolism; Lipopolysaccharides - pharmacology; Mitochondria - metabolism; mitochondria-associated membranes; mitochondrial dysfunction; Protein Serine-Threonine Kinases - metabolism; Proto-Oncogene Proteins c-bcl-2 - metabolism
• ISSN: 0022-0302; 1525-3198
• DOI: 10.3168/jds.2022-22879

Mitochondrial homeostasis is closely associated with cellular homeostasis process, whereas mitochondrial dysfunction contributes to apoptosis and mitophagy. Hence, analyzing the mechanism of lipopolysaccharide (LPS)-caused mitochondrial damage is necessary to understand how cellular homeostasis is maintained in bovine hepatocytes. Mitochondria-associated membranes (MAM), a connection between endoplasmic reticulum (ER) and mitochondria, is important to control mitochondrial function. To investigate the underlying mechanisms of the LPS-caused mitochondrial dysfunction, hepatocytes isolated from dairy cows at ∼160 d in milk (DIM) were pretreated with the specific inhibitors of adenosine 5′-monophosphate-activated protein kinase (AMPK), ER stress, RNA-activated protein kinase-like ER kinase (PERK), inositol-requiring enzyme 1α (IRE1α), c-Jun N-terminal kinase, and autophagy followed by a 12 I1/4g/mL LPS treatment. The results showed that inhibiting ER stress with 4-phenylbutyric acid decreased the levels of autophagy and mitochondrial damage with AMPK inactivation in LPS-treated hepatocytes. The AMPK inhibitor compound C pretreatment alleviated LPS-induced ER stress, autophagy and mitochondrial dysfunction by regulating the expression of MAM-related genes, such as mitofusin 2 (MFN2), PERK, and IRE1α. Moreover, inhibiting PERK and IRE1α mitigated autophagy and mitochondrial dynamic disruption by regulating the MAM function. Additionally, blocking c-Jun N-terminal kinase, the downstream sensor of IRE1α, could reduce the levels of autophagy and apoptosis and restore the balance of mitochondrial fusion and fission by modulating the B cell leukemia 2 (BCL-2)/BCL-2 interacting protein 1 (BECLIN1) complex in the LPS-treated bovine hepatocytes. Furthermore, autophagy blockage with chloroquine could intervene in LPS-caused apoptosis to restore mitochondrial function. Collectively, these findings suggest that the AMPK-ER stress axis is involved in the LPS-caused mitochondrial dysfunction by mediating the MAM activity in bovine hepatocytes.