Protective effects of IL28RA siRNA on cardiomyocytes in hypoxia/reoxygenation injury

Abstract

Objective:We demonstrate the protective effects of the siRNA-mediated inhibition of the interleukin-28 receptor alpha (IL28RA) subunit on cardiomyocytes in hypoxia/reoxygenation (H/R) injury and explore the associated mechanism.Methods:After designing and synthesizing three pairs of siRNA that effectively reduced IL28RA gene expression in vitro (siRNA-6158, siRNA-6160, and siRNA-6162), primary neonatal rat cardiomyocytes were transfected using a liposome transfection method. Six groups were included based on the siRNA that was used and the treatment simulating reperfusion injury: control group, H/R group, H/R+negative control group, H/R+siRNA-6158 group, H/R+siRNA-6160 group, and H/R+siRNA-6162 group. Cell survival and apoptosis rates were measured along with lactate dehydrogenase levels in the cell culture supernatant. Protein levels of IL28RA, phosphatidylinositol 3-kinase, catalytic subunit gamma (PI3KCG), Bcl-2, Bax, and b-actin were also measured.Results:The H/R+siRNA-6158 and H/R+siRNA-6160 groups had significantly higher survival rates and increased PI3KCG-to-b-actin and Bcl-2-to-Bax ratios than the the H/R and H/R+negative control groups (p<0.05). The H/R+siRNA-6158 and H/R+siRNA-6160 groups also exhibited reduced rates of apoptosis and reduced IL28RA-to-b-actin ratios (p<0.05). No significant difference was observed among the H/R+siRNA-6162, H/R, and H/R+negative control groups.Conclusion:IL28RA siRNA-6158 and -6160 were able to protect cardiomyocytes from H/R injury by inhibiting apoptosis. This strategy of inhibiting IL28RA gene expression may reduce reperfusion injury in the treatment of patients with acute myocardial infarction.

Protective effect of celastrol on myocardial ischemia–reperfusion injury

“…H/R process was performed as previously described ( 14 ). Briefly, H9c2 cells were first transferred to the medium without serum and glucose and then placed in a hypoxic chamber saturated with a mixed gas containing 5% CO 2 and 95% N 2 at 37°C for 8 h. Then, cells were reoxygenated by incubating in a fresh medium with serum and glucose under normoxic conditions for 4 h.…”