| 成骨细胞来源的IL-7通过激活mTORC1信号通路抑制骨形成 |
| 投稿时间：2017-03-31 修订日期：2017-04-17 点此下载全文 |
| 引用本文：王华磊.成骨细胞来源的IL-7通过激活mTORC1信号通路抑制骨形成[J].医学研究杂志,2017,46(12):157-160 |
| DOI：
10.11969/j.issn.1673-548X.2017.12.039 |
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| 中文摘要:目的 本研究探讨成骨细胞分泌的白介素-7（IL-7）通过哺乳动物雷帕霉素靶蛋白（mammalian target of rapamycin，mTOR）信号来调节骨形成。方法 将IL-7基因的编码框构建于慢病毒表达载体，导入MC3T3-E1成骨细胞中，应用qRT-PCR和ELISA方法检测IL-7基因mRNA和蛋白的表达效率。Western blot方法检测MC3T3-E1细胞P-S6、S6、Ⅰ型胶原（collagen Ⅰ，Col Ⅰ）以及骨钙素（osteocalcin，Ocn）表达情况。用0.1nmol/L mTORC1特异性抑制剂雷帕霉素刺激过表达IL-7基因的MC3T3-E1成骨细胞，Western blot方法检测P-S6、S6、Col Ⅰ以及Ocn表达情况。结果 过表达IL-7基因的慢病毒载体感染MC3T3-E1细胞可使其IL-7 mRNA表达上调4.6倍，ELISA从蛋白水平证实过表达组细胞上清的IL-7含量增加3.9倍；过表达IL-7的MC3T3-E1细胞P-S6蛋白表达显著增强，而Col Ⅰ和Ocn蛋白表达显著降低；雷帕霉素处理IL-7过表达的MC3T3-E1细胞导致其P-S6蛋白表达显著降低，而Col Ⅰ和Ocn蛋白表达上调。结论 成骨细胞来源的IL-7通过促进mTORC1信号通路抑制其发育成熟。 |
| 中文关键词:成骨细胞 白介素-7 mTOR 骨形成 |
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| Osteoblast-derived IL-7 Prohibits Bone Formation through Activating mTORC1 Signaling |
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| Abstract:Objective To investigate the regulation of bone formation by osteoblast-derived interleukin-7 (IL-7) trough the mammalian target of rapamycin (mTOR) signaling pathway.Methods The lentiviral vector with encoding frame of IL-7 gene was transfected into mouse MC3T3-E1 osteoblasts, and the expression levels of IL-7 mRNA and protein were detected, respectively, by qRT-PCR and ELISA method. Western blot method was used to detect the expression levels of P-S6, S6, collagen Ⅰ (Col Ⅰ) and Osteocalcin (Ocn) in MC3T3-E1 cells and in MC3T3-E1 cells stimulated by 0.1nmol/L mTORC1-specific inhibitor rapamycin.Results The IL-7 mRNA level of MC3T3-E1 cells transfected with upregulated 4.6 times, and ELISA assay showed an increase of 3.9 times of IL-7 level in osteoblastic supernatants. After transfected using lentiviral vector with encoding frame of IL-7 gene, the expression levels of P-S6 protein increased significantly, while the expression levels of Col Ⅰ and Ocn protein decreased significantly. However, rapamycin treatment of over-expressed-IL-7 MC3T3-E1 cells resulted in a decline of P-S6 protein levels, but an increase of Col Ⅰ and Ocn protein levels.Conclusion Osteoblast-derived IL-7 may inhibit its maturation by promoting mTORC1 signaling pathway. |
| keywords:Osteoblast Interleukin-7 mTOR Bone formation |
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