인간 골수유래 중간엽 줄기세포에서 H2O2로 유도한 산화성 손상에 대한 ethyl pyruvate의 세포 보호 효과

Abstract

Pyruvate is an important metabolic intermediate, and also is an effective scavenger of hydrogen peroxide and other reactive oxygen species (ROS). And, pyruvate is relatively unstable in aqueous solutions, which could limit the therapeutic potential of this compound. However, Ethyl pyruvate(EP) is stable and highly soluble agent that makes it convenient as a ROS scavenger. Recent studies have indicated that EP is an effective anti-inflammatory agent in variety of in vivo and in vitro model. In this study, we examined the effect of EP on H2O2 induced oxidative stress in hBM-MSCs by measurement of viability, intracellular ROS, immunohistochemistry and cellular characterization of hBM-MSCs. When comparing with the H2O2 only treated cells, pretreatment with optimal concentration EP (10 μM) after H2O2 induced stress significantly increased cell viability and reduced intracellular ROS production. Also, gene expressions of VEGFA, TGFB1, IGF1, FGF1, BDNF, PIGF, IL4, IL10 were increased by real time PCR analysis. And immunohistochemistry, revealed that expressions of Oct4, Nanog, H-CAM, I-CAM were increased in pretreated hBM-MSC. These results suggest that ethyl pyruvate has cytoprotective effect against H2O2 induced oxidative stress on hBM-MSCs by regulating intracellular ROS. And, there was not any change in hBM-MSCs characterization. Therefore, ethyl pyruvate pretreatment in hBM-MSCs culture process would be safe strategies for preventing inevitable oxidative stress happened in routine culture process.