阿魏酸抗成骨细胞凋亡作用及其GPR30相关性研究

黄天一, 陈婷婷, 崔杰, 李梦雨, 华永庆

黄天一, 陈婷婷, 崔杰, 李梦雨, 华永庆. 阿魏酸抗成骨细胞凋亡作用及其GPR30相关性研究[J]. 南京中医药大学学报, 2020, 36(3): 346-351.
引用本文: 黄天一, 陈婷婷, 崔杰, 李梦雨, 华永庆. 阿魏酸抗成骨细胞凋亡作用及其GPR30相关性研究[J]. 南京中医药大学学报, 2020, 36(3): 346-351.
HUANGTian-yi, CHENTing-ting, CUIJie, LIMeng-yu, HUAYong-qing. Study on the Anti-Apoptosis of Ferulic Acid on Osteoblast and Its Correlation with GPR30[J]. Journal of Nanjing University of traditional Chinese Medicine, 2020, 36(3): 346-351.
Citation: HUANGTian-yi, CHENTing-ting, CUIJie, LIMeng-yu, HUAYong-qing. Study on the Anti-Apoptosis of Ferulic Acid on Osteoblast and Its Correlation with GPR30[J]. Journal of Nanjing University of traditional Chinese Medicine, 2020, 36(3): 346-351.

阿魏酸抗成骨细胞凋亡作用及其GPR30相关性研究

Study on the Anti-Apoptosis of Ferulic Acid on Osteoblast and Its Correlation with GPR30

  • 摘要: 目的 观察阿魏酸(FA)对成骨细胞凋亡的影响及其机制,探讨FA对骨质疏松的潜在治疗作用。方法 选择前成骨细胞系MC3T3-E1,MTT法观察FA对细胞活力的影响;流式细胞术检测细胞凋亡;Western blot法检测凋亡相关蛋白Bcl-2的表达;流式细胞术及DCFH-DA荧光染色观察活性氧(ROS)水平;免疫荧光法观察叉头框蛋白(Fox)家族FoxO3a的核定位;采用G蛋白偶联受体GPR30特异性拮抗剂G15进行干预,观察FA效应的变化。结果 FA在25~400 μmol/L范围内对正常细胞无显著影响,但对过氧化氢(H2O2)损伤后的细胞活力有促进作用;流式结果表明,FA可以减少过氧化损伤后的细胞凋亡率和降低ROS水平;免疫荧光结果表明,FA能阻碍氧化应激条件下细胞凋亡和氧化应激相关蛋白FoxO3a的核内含量。G15处理后,FA引起的抗细胞凋亡、降低ROS水平及FoxO3a核内含量减少作用均被显著阻断。结论 FA具有抗成骨细胞凋亡、降低细胞内ROS水平、阻止过氧化损伤条件下FoxO3a核易位的作用,其机制可能与膜雌激素受体GPR30有关。
    Abstract: OBJECTIVE To investigate the effects of ferulic acid (FA) on osteoblast apoptosis and its mechanism, and explore the potential therapeutic effects of FA on osteoporosis. METHEODS The pre-osteoblast cell line MC3T3-E1 was selected. The effect of FA on cell viability was observed by MTT assay. Apoptosis was detected by flow cytometry. The expression of apoptosis-related protein Bcl-2 was detected by Western blot. Flow cytometry and DCFH-DA fluorescence staining were performed to observe the reactive oxygen species (ROS) level. The nuclear localization of FoxO3a, one member in the forkhead box (Fox) family, was observed by immunofluorescence. The G protein coupled receptor 30 (GPR30) specific antagonist G15 was used to observe FA effect. RESULTS FA showed no significant effect on normal cells in the dose range of 25~400 μmol/L, but promoted the cell viability after hydrogen peroxide injury. Flow results showed that FA could reduce cell apoptosis rate and ROS level after peroxidation injury. Immunofluorescence results indicated that FA could inhibit the apoptosis and the nuclear content of oxidative stress-related protein FoxO3a under peroxidative conditions. After treatment with G15, the effects of FA on decreasing apoptosis, lowering ROS levels, and reducing the content of FoxO3a in the nucleus were significantly blocked. CONCLUSION FA shows the ability to resist osteoblast apoptosis, reduce intracellular ROS levels, and prevent FoxO3a nuclear translocation under conditions of peroxidative damage. The mechanism may be related to the membrane estrogen receptor GPR30.
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