Song Jae-Yen, Kim Min-Joung, Jo Hyun-Hee, Hwang Seong-Jin, Chae Boah, Chung Jae-Eun, Kwon Dong-Jin, Lew Young-Ok, Lim Yong-Taik, Kim Jang-Heub, Kim Jin-Hong, Kim Mee-Ran
Department of Obstetrics and Gynecology, The Catholic University of Korea, Republic of Korea.
J Steroid Biochem Mol Biol. 2009 Oct;117(1-3):74-80. doi: 10.1016/j.jsbmb.2009.07.006. Epub 2009 Jul 25.
This study discussed the role of estrogen as an antioxidant in the damage of vascular endothelial cells.
We treated bovine aortic endothelial cells (bAEC) either with 1mM of H(2)O(2) alone or with 1 microM of 17beta-estradiol (E(2)) for 24h followed by 1mM of H(2)O(2) for 3h. The cell survival was evaluated by MTT assay, cellular apoptosis by fluorescence activated cell sorter (FACS) and Hoechst 33342 staining, oxidative stress by intracellular reactive oxygen species (ROS) and apoptosis after oxidative stress by western blotting for phospho-p38, p38, and Bcl-2.
MTT assay showed that bAEC viability was reduced to 55.7+/-3.0% and 39.1+/-3.7% after 30 and 60 min of H(2)O(2) treatment, respectively. E(2) and H(2)O(2) treated cells did not show significant decrease in the cell survival. Similarly the FACS analysis and Hoechst 33342 stain showed that the latter decreased cellular apoptosis induced by H(2)O(2). Intracellular ROS increased by 181.6+/-68.9% in the former and by 37.0+/-3.9% in the latter (P<0.05). The expression of phospho-p38 mitogen-activated protein kinase (MAPK) was higher in the latter.
E(2) mediates antioxidant effects on the oxidative stress induced by H(2)O(2). This antioxidant effect on bAEC may elucidate the scientific basis of hormone therapy for maintaining cardiovascular integrity in postmenopausal women.
本研究探讨雌激素作为抗氧化剂在血管内皮细胞损伤中的作用。
我们将牛主动脉内皮细胞(bAEC)单独用1mM过氧化氢(H₂O₂)处理,或先用1μM 17β-雌二醇(E₂)处理24小时,随后再用1mM H₂O₂处理3小时。通过MTT法评估细胞存活率,用荧光激活细胞分选仪(FACS)和Hoechst 33342染色评估细胞凋亡,用细胞内活性氧(ROS)评估氧化应激,并用蛋白质免疫印迹法检测磷酸化p38、p38和Bcl-2来评估氧化应激后的细胞凋亡。
MTT法显示,H₂O₂处理30分钟和60分钟后,bAEC活力分别降至55.7±3.0%和39.1±3.7%。E₂和H₂O₂处理的细胞在细胞存活率上没有显著下降。同样,FACS分析和Hoechst 33342染色显示,后者可减少H₂O₂诱导的细胞凋亡。前者细胞内ROS增加了181.6±68.9%,后者增加了37.0±3.9%(P<0.05)。后者磷酸化p38丝裂原活化蛋白激酶(MAPK)的表达更高。
E₂介导了对H₂O₂诱导的氧化应激的抗氧化作用。这种对bAEC的抗氧化作用可能阐明了激素疗法维持绝经后女性心血管完整性的科学依据。