| 摘要: |
| 目的探讨17β-雌二醇对成骨细胞中维生素D受体(vitamin D receptor, VDR)表达的调节，以及丝裂原激活的蛋白激酶(motigen-activated protein kinase, MAPK)信号通路是否参与该过程。方法小鼠前成骨细胞MC3T3-E1亚克隆14用添加10% 小牛血清的无酚红α-MEM培养液培养,在干预细胞时,换为不含血清的无酚红α-MEM培养液。用实时荧光定量RT-PCR法和蛋白质印迹法检测17β-雌二醇干预前后细胞中VDR基因和蛋白的表达,用蛋白质印迹法检测17β-雌二醇干预前后细胞中MAPK信号通路的激活。然后用MAPK信号通路阻断剂U0126和17β-雌二醇同时处理细胞,观察VDR表达的变化。结果17β-雌二醇作用72 h可以上调MC3T3-E1细胞中VDR基因和蛋白的表达；17β-雌二醇可在15 min内激活ERK信号通路,并持续到60 min,但是不能激活JNK和p38信号通路；应用U0126后,17β-雌二醇对成骨细胞中VDR表达水平的上调作用可被部分抑制。 结论17β-雌二醇可上调成骨细胞中VDR的表达,并能快速激活成骨细胞中MAPK信号通路;MAPK信号通路参与了雌激素上调VDR表达的过程。 |
| 关键词: 雌二醇 维生素D受体 成骨细胞 丝裂原活化蛋白激酶类 |
| DOI：10.3724/SP.J.1008.2011.01291 |
| 投稿时间：2011-05-25修订日期：2011-10-15 |
| 基金项目:国家自然科学基金(30872726). |
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| MAPK signal pathway is involved in 17β-estrodial-induced upregulation of vitamin D receptor in osteoblasts |
| ZHOU Yun,ZHANG Xiu-zhen,SONG Li-ge* |
(Department of Endocrinology, Tongji Hospital, Tongji University School of Medicine, Shanghai 200065, China
*Corresponding author.) |
| Abstract: |
| ObjectiveTo explore the regulatory effect of 17β-estrodial on vitamin D receptor (VDR) expression in pre-osteoblasts and the involvement of MAPK signal pathway in the process. MethodsMC3T3-E1 subclone 14 cells were cultured in phenol-red free medium supplemented with 10% fetal bovine serum (FBS). Serum free medium was used when the cells were experimentally treated. After the cells were treated with 17β-estrodial for pre-determined time periods, expressions of vitamin D receptor (VDR) mRNA and protein were determined by SYBR green-based quantitative PCR and Western blotting analysis, respectively; and the activation of MAPK in MC3T3-E1 cells was examined by Western blotting analysis. Then the changes of VDR mRNA and protein in MC3T3-E1 cells were detected after the cells were treated with MAPKs inhibitors and 17β-estrodial. ResultsVDR mRNA and protein were upregulated in MC3T3-E1 cells after treatment with 17β-estrodial for 72 h. ERK/MAPK signal in MC3T3-E1 cells was activated within 15 min after treatment with 17β-estrodial and the activation remained for 60 min; but it did not activate JNK and p38 MAPK pathways. 17β-estrodial-induced VDR upregulation in MC3T3-E1 cells could be partly inhibited by ERK/MAPK inhibitor U0126. Conclusion17β-estrodial can upregulate VDR expression in osteoblasts and can rapidly activate MAPK signal pathway, which is involved in the estrogen-induced upregulation of VDR. |
| Key words: estrodiol vitamin D receptors osteoblasts mitogen-activated protein kinases |
