Pterostilbene attenuates oxidative stress induced by hydrogen peroxide in MAC-T cells through activating PINK1/Parkin-mediated mitophagy

Abstract

High‐producing dairy cows are susceptible to altered redox balance owing to their high secretion and strong metabolism during transition to lactation. Previously reports demonstrated that pterostilbene (PTE) alleviate hydrogen peroxide (H₂O₂)-induced oxidative damage. However, molecular mechanism underlying protective role of PTE in H₂O₂-elicited oxidative stress in bovine mammary epithelial cells (MAC-T cells) remains unclear. The aims of our research is to clarify the potential molecular mechanisms of PTE in H₂O₂-triggered oxidative damage. MAC-T cells were pre-treated with PTE (0, 10, 25 and 50 μM) for 12 h and then cultured with H₂O₂ (0, 100, 200, 400, 600, 800 and 1000 μM) for another 24 h. The results were interpreted by CCK8, western blot, and immunofluorescence assay. Results showed that PTE significantly attenuated H₂O₂-mediated reduction in T-AOC, SOD and GSH-Px activity. Mechanistic studies found that PTE restricted the H₂O₂-induced protein expression of HO-1, GPX4 and SOD1, and PTE reversed intracellular MDA and ROS generation and decreased the MMP in MAC-T cells. Moreover, H₂O₂ exposure markedly inhibited mitophagy, whereas PTE pre-treatment activated PINK1/Parkin-mediated mitophagy pathway, which further limited ROS production. Overall, PTE can be used to improve oxidative stress and as a novel antioxidant agent in dairy cows.

Highlights

• PTE restored the levels of H₂O₂-induced oxidative stress markers (T-AOC, SOD, GSH-Px and MDA) in MAC-T cells.

• PTE effectively blocked the increase of protein expression (HO-1, SOD1 and GPX4) in H₂O₂-induced MAC-T cells.

• PTE remarkably decreased the accumulation of ROS and improved MMP in H₂O₂-induced MAC-T cells.

• PTE exerted a significant protective effect against oxidative stress injury by reducing the inhibition of H₂O₂ on mitophagy.

Keywords:

• Pterostilbene
• oxidative stress
• PINK1/Parkin
• mitophagy
• MAC-T

Ethical approval

All procedures were approved by the Animal Care and Use Committee of Xuzhou University of Technology, Xuzhou, China.

Disclosure statement

No potential conflict of interest was reported by the author(s).

Data availability statement

The original data of the paper are available upon request from the corresponding author.

Additional information

Funding

The present study were supported by the Natural Science Foundation of Jiangsu Province [BK20210077, BK20211051, and BK20210078], the Natural Science Foundation of the Jiangsu Higher Education Institutions of China [22KJB230008 and 22KJA240003] and Key Research and Development Plan of Xuzhou City [KC23034, KC21296 and KC22133].