| Summary: | 碩士 === 國立臺灣大學 === 藥理學研究所 === 101 === Background: Ischemic heart disease is one of the most common heart diseases worldwide. Although restore blood flow after acute coronary artery occlusion is inevitable, reperfusion injury might cause more severe damage to the heart. This study examines whether the flavonoid-like compound, 36-13, has cardioprotective effects under oxidative stress, and investigates the possible underlying mechanism.
Methods and Results: This study used 50μM H2O2 to induce oxidative stress of NRVM and H9C2 cell line. We first used MTT assay to observe the cardioprotective effect of 36-13, compared the effective concentration with antioxidant NAC, and then compared their influence on the intracellular ROS. Next, the activation of different proteins ( eNOS, AMPK, Akt, and ERK ) at different time points under oxidative stress were shown by western blot. Then we pretreated different protein inhibitors to investigate the importance of different protein on cell viability, and followed by western blot to investigate detail mechanism. The result of MTT assay shows that 3μM 36-13 and 50μM NAC can enhance the cell viability significantly under oxidative stress, but 36-13 can’t reduce intracellular ROS induced by H2O2 as NAC. Western blot shows that 36-13 can enhance expressions of peNOS, pAMPK, and pAkt under oxidative stress. The following MTT assay reveals that inhibition of eNOS and AMPK phosphorylation can attenuate the protective effect of 36-13 significantly. The result of further western blot study indicates that the activation of eNOS caused by 36-13 was regulated by AMPK.
Conclusion: 36-13 reveals protective effects to cardiomyocytes under oxidative stress, not through antioxidant or reduces intracellular ROS generation, but activates AMPK to regulate eNOS.
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