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[Y-27632 protects H9c2 cardiomyocytes against hypoxia/reoxygenation injury by inhibiting autophagy pathway].

Objective To investigate the protective effect of preconditioning with Rho-kinase inhibitor Y-27632 on H9c2 cardiomyocytes against hypoxia/reoxygenation (H/R) injury, and explore its underlying mechanism. Methods H9c2 cardiomyocytes were divided into normal group, H/R group, Y-27632 group and Y-27632 plus 3-methyladenine (Y-27632/3-MA) group. The cells of the normal group were cultured with 950 mL/L air and 50 mL/L CO2 at 37DegreesCelsius for 6 hours. The cells of the H/R group were pretreated with hypoxia (940 mL/L N2, 10 mL/L O2, 50 mL/L CO2) for 3 hours, followed by 3-hour reoxygenation (950 mL/L air, 50 mL/L CO2). The cells of the Y-27632 group were preconditioned with 10 μmol//L of Rho-kinase inhibitor Y-27632 for 1 hour followed by H/R treatment as the H/R group did. The cells in the Y-27632/3-MA group were preconditioned with 10 μmol//L of Y- 27632 and 5 mmol/L of autophagy inhibitor 3-MA for 1 hour before H/R treatment. After the treatments, CCK-8 assay was used to detect cell survival rate and chemical colorimetry was used to detect lactate dehydrogenase (LDH) activity. Cell apoptosis was measured by flow cytometry. The protein levels of phosphorylation myosin light chain (p-MLC) and beclin 1 were analyzed by Western blot analysis. Results After Y-27632 pretreatment, the survival rate of H9c2 cells were improved; the LDH activity, the apoptosis rate and the protein level of p-MLC were reduced; and the level of beclin 1 was raised. However, in the Y-27632/3-MA group, the anti-apoptotic effect on H9c2 cells was significantly attenuated, the protein level of p-MLC increased and the level of beclin 1 decreased. Conclusion Preconditioning with Y-27632 can alleviate H/R-induced myocardium injury through inhibiting Rho-kinase and promoting autophagy pathway.