Gomisin A inhibits hypoxia/reoxygenation induced myocardial cell injury by modulating TLR4 NF κB pathway

Abstract

Purpose: To determine the impact of Gomisin A (GomA) on myocardial cell damage caused by hypoxia/reoxygenation (H/R). Methods: Various methods, including MTT, western blot and flow cytometry were used to to assess the viability of H9c2 cardiomyocytes and cell apoptosis. Expression levels of various enzymes and cytokines, including superoxide dismutase (SOD), malondialdehyde (MDA), catalase (CAT), tumor necrosis factor-alpha (TNF-α), interleukin (IL)-1β and IL-6 were measured by enzyme-linked immunosorbent assay (ELISA). Western blot was used to evaluate the expression levels of inducible nitric oxide synthase (iNOS), cyclooxygenase 2 (COX-2) and Toll-like receptor 4 (TLR4), as well as the phosphorylation of p65 and nuclear factor kappa B (NF-κB) inhibitor alpha (IκBα). Results: Low concentrations of GomA had no toxic effect on H9c2 cells. In H/R-stressed H9c2 cells, GomA increased cell viability and reduced cell apoptosis, suggesting that GomA inhibited H/R-induced cell apoptosis. Additionally, GomA alleviated H/R-induced oxidative stress and inflammation by increasing the expression of SOD and CAT, while decreasing the expression of MDA, iNOS, COX-2, TNF-α, IL-1β, and IL-6. GomA also suppressed the H/R-induced TLR4–NF-κB pathway by reducing the expression of TLR4 and the phosphorylation of p65 and IκBα. Conclusion: These results indicate that GomA is a potential candidate for the treatment of myocardial ischemia/reperfusion (MI/R) injury by increasing cell viability, reducing cell apoptosis, alleviating oxidative stress, and reducing inflammation by inhibiting TLR4–NF-κB pathway.