Effect of exogenous spermine on hypoxia-induced cardiomyocyte apoptosis in suckling rats and its mechanism

doi:10.12047/j.cjap.2646.2018.046

Abstract

Abstract: Objective: To observe the effect of exogenous spermine on H/I-induced cardiomyocyte apoptosis in suckling rat, and to explore its mechanism.Methods: The hypoxia injury model was established on primarily cultured suckling rat cardiomyocytes (Hank's balanced salt solution (pH=6.8) was used as cell culture medium, the oxygen in culturing environment was discharged, and the suckling rat cardiomyocytes were cultured in an anaerobic incubator for 24 hours). Cultured suckling rat cardiomyocytes were randomly assigned into control, hypoxia and hypoxia+spermine (Hypoxia+Sp) groups. The protein expressions of key enzyme catalyzing polyamine metabolism (ODC, SSAT) in cardiomyocytes were detected by Western blot. The cell apoptosis was detected by using Cell Counting Kit-8 (CCK-8) and Hoechst 33342 dye. The activities of total-superoxide dismutase (SOD), caspase-3/-9 and the contents of malondialdehyde (MDA), glutathione (GSH) in cell medium were analyzed. Intracellular reactive oxygen species (ROS) generation was estimated by using DCFH-DA staining.Results: Compared with the control group, the expression of SSAT protein, apoptosis rate, MDA content, the activity of caspase-3/-9 and ROS generation were increased, but the expression of ODC protein, SOD activity and GSH content were decreased in hypoxia group. Compared with the hypoxia group, the changes of all index above-mentioned were weakened in the Hypoxia+Sp group.Conclusion: Exogenous spermine can attenuate cardiomyocyte injury and cell apoptosis of suckling rat induced by hypoxia,which mechanism is related with restoring polyamines homostasis and scavenging ROS.

Key words: spermine, cardiomyocytes, ROS, apoptosis, hypoxia, suckling rat

YUAN Di, WANG Yue-hong, YUAN Hui, SHAO Yi-ying, FAN Yu-qi, SHAO Xiao-ting, HU Jing, WEI Can, XU Chang-qing. Effect of exogenous spermine on hypoxia-induced cardiomyocyte apoptosis in suckling rats and its mechanism[J]. CJAP, 2018, 34(3): 193-196.

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