Swine acute diarrhoea syndrome coronavirus (SADS-CoV) is an enveloped, single-stranded positive-sense RNA virus that causes acute diarrhoea and death in piglets, resulting in significant economic losses to the pig farming industry. Studying the interaction patterns between SADS-CoV and host proteins can provide guidance for the development of antiviral drugs. In previous work, we identified 289 host proteins interacting with the SADS-CoV M protein through glutathione S-transferase pull down combined with LC-MS/MS. Here, we focus on prohibitin (PHB), which is associated with the stability of mitochondrial function in cells, and demonstrate that there is a physical interaction and cellular co-localization relationship between the SADS-CoV M protein and PHB protein. Additionally, SADS-CoV-mediated infection has a strong correlation with PHB expression, and regulating PHB expression dose-dependently antagonizes SADS-CoV replication. Moreover, we discovered that PHB has an antagonistic effect on apoptosis induced by SADS-CoV infection. Overall, this work helps to elucidate the role of the PHB protein in the SADS-CoV life cycle, providing a potential target for antiviral research.