| Summary: | 碩士 === 臺灣大學 === 藥理學研究所 === 98 === Background：Reperfusion injury during recovery phase of tissue ischemia is important in many clinical situations such as cardiac transplantation, myocardial infarction and stroke, which influence outcome of the patients. We are interested in whether the natural ingredients and anti-diabetic medicines are protective against ischemic heart disease. The aim of the present study was to investigate the protective effects of flavonoids and metformin on H9c2 hypoxia-reoxygenation induced injury.
Methods and Results：We used rat heart-derived cell line, H9c2, for the experiments. Cells were placed in hypoxia chamber with no glucose no serum medium to induce hypoxia 6 hr, then changed to normoxic condition for 12 hours. Cells were divided into vehicle and drug group. In drug group, RS47, 642D and metformin were added in the medium 1 hr before hypoxia. MTT assay was used to show cell viability of hypoxia reoxygenation. Western blot、PI3K-Akt inhibitor LY294002、ERK inhibitor PD98059 and AMPK inhibitor Compound C were also used in the present study. In MTT assay, cell viability was significantly increased in the drug group when compared to sham group. Western blot analysis revealed that p-Akt、p-AMPK and p-ERK were increased in the drug group, p-p38 and Bax were decreased, and p-GSK-3β was elevated. The use of Compound C significantly inhibited the protective effect of RS47、642D and metformin. We also measured LDH release, the results matched MTT assay.
Conclusion：RS47, 642D and metformin signigicantly increased the cell viability in H9c2 cell hypoxia reoxygenation injury, and this was attributed to a decrease of cell apoptosis and necrosis. The decreased reperfusion injury was related to activation of Akt、AMPK and ERK.
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