Objective To investigate the effects of increased expression of Omi/HtrA2 on myocardial autophagy in aging myocardiocytes. Methods The rat embryonic myocardial cell line H9c2 was induced by D-galactose (8g/L) to establish aging model. Cell aging was detected by β-galactosidase staining. Cell viability was detected by CCK8 kits and lactic dehydrogenase (LDH) activity analysis. Ucf-101, a specific inhibitor of Omi/HtrA2, was used to inhibit the activity of Omi/HtrA2, and the H9c2 cells with stable transfection of Omi/HtrA2 was constructed to increase the expression of Omi/HtrA2. The expression of Omi/HtrA2, beclin1 and LC3-Ⅱ were detected by Western blotting. Results (1) D-galactose resulted in a significant increase in positive staining to β-galactosidase in the induced cardiacmyocytes than the H9c2 cells [(87.7±3.60)% vs (918.3±2.80)%, P＜0.01]. The cck8 results showed that there was no significant difference between the cells with and without D-galactose inducement (P＞0.05). LDH activity was increased in the induced H9c2 cells than those without (7.07±0.65 vs 5.93±3.36, P＜0.01). (2) The expression of Omi/HtrA2 was increased (P＜0.05), but the expression of beclin1 was decreased (P＜0.01) in cells induced by D-galactose, as compared with H9c2 cells. In cells treated with ucf-101, the expression of Omi/HtrA2 was significantly decreased (P＜0.05), but the expression of beclin1 was further decreased (P＜0.01).（3）The expression of LC3-Ⅱwas significantly increased in Omi/HtrA2-overexpressed cells, as compared with H9c2 cells (P＜0.05). But, ucf-101 resulted in the expression of LC3-Ⅱdecreased in the Omi/HtrA2-overexpressed cells (P＜0.05). Conclusion Increased expression of Omi/HtrA2 promotes autophagy in aging myocardiocytes.