Objective: The present study was designed to investigate whether mitochondrial dysfunction and oxidative stress in HTN-injured cardiomyocytes are associated with SIRT3 ubiquitination and whether SIRT3 deubiquitination can ameliorate the related cardiomyocyte injury. Design and method: The human cardiomyocyte AC16 cell line was selected as the study target. The SIRT3-K288R cell line construction was verified by immunoprecipitation and the effect of Ang-II on SIRT3 class ubiquitination was explored; the expression of SOD2, Acetyl-SOD2 (MnSOD), protein acetylation level, inflammatory vesicles NLRP3, etc. in different subgroups were determined by Western Blot to explore the ubiquitination-like modifications and SIRT3 deacetylation in HTN-related cell injury Results: (1) Immunoprecipitation results showed that the level of SIRT3 class ubiquitination was reduced after SIRT3-K288R transfection; compared to the control group, the level of SIRT3 class ubiquitination was increased in the presence of AngII; (2) Western Blot results showed that compared to the HTN cell injury model, the cardiomyocytes in the SIRT3-K288R intervention group had increased Acetyl- SOD2/SOD2 levels were increased (p < 0.05) and protein acetylation levels were decreased in the SIRT3-K288R intervention group compared with the HTN cell injury model, indicating that SIRT3 deubiquitination increased the deacetylation effect on mitochondria and increased cellular antioxidant stress enzyme activity; (3) Western Blot results showed that compared with the HTN cell injury model, cardiomyocytes in the SIRT3-K288R intervention group NLRP3 and NOX4 levels were significantly reduced, indicating that SIRT3 deubiquitination could improve the inflammatory response to Ang-II-related cell injury. Conclusions: In the HTN-associated cell injury model, the level of SIRT3 class ubiquitination was significantly increased, and the removal of ubiquitination modification by viral transfection could improve oxidative stress, protein acetylation levels and associated inflammatory responses in HTN-injured cardiomyocytes, indicating that SIRT3 class ubiquitination may play an important mechanism in HTN-associated cell injury, and reducing SIRT3 class ubiquitination modification may be a possible direction for future research.