Protective Effect of Fasudil on Hydrogen Peroxide-Induced Oxidative Stress Injury of H9C2 Cardiomyocytes

OBJECTIVE: Oxidative damage is a pathological factor that causes cardiovascular damage in the clinic and is increasingly serious. This study focused on the effect of fasudil on H(2)O(2)-induced oxidative damage in cardiomyocytes. MATERIALS AND METHODS: H9C2 cardiomyocytes were cultured in vitro and divided into three groups: control group (Con group), H(2)O(2) treatment (H(2)O(2) group), and fasudil and H(2)O(2) cotreatment (H(2)O(2)+fasudil group). The content levels of LDH and MDA in the supernatant were detected, and the morphology of H9C2 cardiomyocytes was observed by light microscopy. 8-OHdG staining was observed by a fluorescence inversion microscope. Cell Counting Kit (CCK-8), western blotting, real-time polymerase chain reaction (RT-PCR), and enzyme-linked immunosorbent assay (ELISA) were used to investigate the effect of fasudil on the Rho/ROCK signaling pathway. RESULTS: Our results showed that after H(2)O(2) treatment, the H9C2 cardiomyocytes were irregular in shape and elliptical. But the morphology of the H(2)O(2)+fasudil group was similar to that of the Con group. The green fluorescence of the H(2)O(2) group was significantly enhancer than that of the Con group, while the green fluorescence of the H(2)O(2)+fasudil group was weaker than those of the H(2)O(2) group. By detecting the supernatant, it was found that the contents of LDH were significantly increased, and the contents of SOD and CAT in the H(2)O(2) group were significantly decreased. And the expression of antioxidant indicators in the H(2)O(2) group was significantly decreased by western blotting. The results of RT-PCR showed that SOD1 and SOD2 mRNA in the H(2)O(2) group was significantly reduced, and the contents of GPX1 and GPX3 in the H(2)O(2) group were significantly decreased by enzyme-linked immunosorbent assay (ELISA). The expression of ROCK1, ROCK2, and downstream phosphorylation of myosin phosphatase target subunit-1 (p-MYPT-1) was significantly increased in the H(2)O(2) group, while fasudil inhibited the increase of ROCK1, ROCK2, and p-MYPT-1. CONCLUSIONS: Fasudil can inhibit the Rho/ROCK signaling pathway induced by H(2)O(2) and reduce oxidative stress response, inhibit apoptosis, and improve antioxidant enzyme activity in H9C2 cardiomyocytes thereby delaying cell senescence.