Abstract：Objective To investigate the effect of Isoflurane (Iso) on anoxia/reoxygenation (A/R)-induced cardiocyte injury and underlying mechanism. Methods Cultured cardiocytes (H9c2) were pre-treated with Iso (1%, 2%, 3%) and then stimulated with 3 h of anoxia and 3 h of reoxygenation. Cell proliferation was measured by MTT method, and the Iso concentration was selected. The miR-499 overexpression reagent, miR-499 mimic was transfected into cardiocytes pre-treated with 3% Iso and simulated with anoxia/reoxygenation. Cell apoptosis was detected by flow cytometry assay. The expression level of caspase-3, caspase-9, Bax and Bcl-2 were measured by Western blot. The contents of lactic dehydrogenase (LDH), creatine kinase (CK) and alanine aminotransferase (ALT) were detected by corresponding kit, respectively. Results Compared with the control group, cell proliferation was significantly decreased in A/R group (P ＜ 0.05), and increased dose-dependently in Iso pre-treated cells (P ＜ 0.05), thus 3% was selected to be the concentration of Iso pre-treatment. Compared with the control group, the expression of miR-499 was significantly increased in A/R group (P ＜ 0.05), and decreased in 3% Iso treated cells (P ＜ 0.05). Compared with the control group, cell proliferation was significantly decreased (P ＜ 0.05); cell apoptosis was increased (P ＜ 0.05); the expression of pro-apoptotic protein caspase-3, caspase-9 and Bax were increased (P ＜ 0.05), anti-apoptotic protein Bcl-2 was decreased (P ＜ 0.05); the content of LDH, CK and AST were increased in A/R group (P ＜ 0.05). Iso pre-treat inhibited A/R induced H9c2 injury, and the protective effect of Iso was reversed by miR-499 overexpression in damaged cardiocyte. Conclusion Isoflurane attenuates anoxia/reoxygenation-induced cardiocyte injury through inhibiting miR-499 expression.
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2018, Vol. 15 
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