| 张津铭,吕正涛,卢伟伟,等.基质细胞衍生因子-1对小鼠关节软骨细胞自噬水平的影响.骨科,2016,7(4): 268-273. |
| 基质细胞衍生因子-1对小鼠关节软骨细胞自噬水平的影响 |
| Effect of stromal derived factor-1 on autophagy level of articular chondrocyte in mice |
| 投稿时间：2015-12-26 |
| DOI：10.3969/j.issn.1674-8573.2016.04.011 |
| 中文关键词: 基质细胞衍生因子-1 软骨细胞 自噬 骨关节炎 |
| 英文关键词: Stromal cell-derived factor-1 Chondrocyte Autophagy Osteoarthritis |
| 基金项目:国家自然科学基金（81472082，81572094） |
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| 中文摘要: |
| 目的 通过研究基质细胞衍生因子-1（stromal derived factor-1, SDF-1）对小鼠关节软骨细胞自噬的影响，探讨SDF-1在骨性关节炎中的作用及机制。方法 分离并体外培养小鼠关节软骨细胞，分别予以0、1、10、100、1 000 μg/L浓度的五组重组SDF-1蛋白干预24 h。运用RT-PCR检测各组细胞的微管相关蛋白1轻链3（microtubule associated protein 1 light chain 3, LC3）和UNC-51样激酶复合物1（uncoordinated-51 like kinase 1, ULK1）mRNA表达情况，运用Western Blot方法检测LC3-Ⅱ、LC3-Ⅰ、ULK1及泛素连接蛋白62（ubiquitin-binding protein p62, p62）蛋白表达水平，通过透射电镜观察自噬溶酶体。结果 RT-PCR结果显示10、100、1 000 μg/L浓度条件下LC3、ULK1的mRNA水平明显高于对照组，且差异具有统计学意义（P＜0.05）。Western Blot结果显示1、10、100、1 000 μg/L的各组细胞的LC3-Ⅱ/LC3-Ⅰ比值、ULK-1表达水平较对照组升高，p62蛋白表达水平较对照组明显降低，差异具有统计学意义（P＜0.05）。透射电镜结果显示经SDF-1干预后，自噬溶酶体较对照组增多，差异具有统计学意义（P＜0.05）。结论 SDF-1可诱导小鼠关节软骨细胞自噬的发生，SDF-1可能通过调节软骨细胞自噬水平参与了骨性关节炎的进展。 |
| 英文摘要: |
| Objective To investigate the effect of stromal derived factor-1 (SDF-1) on autophagy level of articular chondrocyte in mice and to explore the effect and mechanism of SDF-1 in osteoarthritis. Methods The articular chondrocytes were obtained from 3- to 4-days-old C57BL/6 mice, cultured in vitro, and treated by recombinant SDF-1 protein for 24 h. The concentrations of SDF-1 were 0, 1, 10, 100 and 1 000 μg/L respectively. The mRNA levels of LC3 and ULK1 were detected by the real-time polymerase chain reaction (RT-PCR). The expression levels of LC3-Ⅱ, LC3-Ⅰ,ULK1 and p62 proteins were examined by Western Blot. Results After chondrocytes were treated with SDF-1 of 10, 100 and 1 000 μg/L, the mRNA expression levels of LC3 and ULK1 were up-regulated as compared with control group. The ratio of LC3-Ⅱ to LC3-Ⅰ and the expression of ULK-1 protein were increased, and the expression of p62 protein was reduced in SDF-1-treated groups (1, 10, 100 and 1 000 μg/L) as compared with control group (P＜0.05 for all). Transmission electron microscopy revealed that as compared with control group, autophagic lysosomes were increased significantly after chondrocytes were treated with SDF-1 (P＜0.05). Conclusion SDF-1 can induce autophagy of articular chondrocytes in mice. SDF-1 maybe participate the progress of osteoarthritis through regulating autophagy level of articular chondrocyte. |
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