| 向威,许涛.初级纤毛在调控软骨细胞自噬中的作用研究.骨科,2018,9(6): 474-478. |
| 初级纤毛在调控软骨细胞自噬中的作用研究 |
| Role of primary cilia in the regulation of autophagy in chondrocytes |
| 投稿时间：2018-08-26 |
| DOI：DOI：10.3969/j.issn.1674-8573.2018.06.011 |
| 中文关键词: 软骨 初级纤毛 自噬 |
| 英文关键词: Chondrocyte Primary cilia Autophagy |
| 基金项目:国家自然科学基金（81772440） |
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| 中文摘要: |
| 目的 探究初级纤毛和自噬在软骨细胞中的表达关系，以及调控初级纤毛表达对软骨细胞自噬的影响。方法 首先通过免疫荧光染色检测软骨细胞中自噬小体和初级纤毛的表达水平及二者在软骨细胞分裂周期中的定位表达关系；通过使用无血清培养基诱导初级纤毛表达，水合氯醛破坏纤毛结构后，统计分析初级纤毛发生率、长度和自噬的表达，Western blot法检测初级纤毛和自噬相关蛋白IFT88、Beclin1和LC3-Ⅰ/Ⅱ的表达情况。结果 软骨细胞中存在初级纤毛和自噬的共表达，初级纤毛的表达率为（62.16±11.74）%，长度为（2.342±0.392） μm，单个细胞内自噬的相对表达量为26.38±5.31；在软骨细胞分裂周期中，自噬小体的数量、大小和分布会随着初级纤毛的解聚和重构发生变化；无血清培养基可诱导初级纤毛表达率上调，促进纤毛延长，同时会上调软骨细胞自噬水平及IFT88、Beclin1和LC3-Ⅱ的表达，而水合氯醛破坏纤毛结构后，初级纤毛表达率和长度下调，细胞自噬表达下调，同时抑制IFT88、Beclin1和LC3-Ⅱ蛋白的表达。结论 在软骨细胞中存在初级纤毛和自噬的定位表达关联性，调控初级纤毛的表达会影响软骨细胞的自噬水平。 |
| 英文摘要: |
| Objective To explore the expression relationship between primary cilia and autophagy in chondrocytes, and to detect the effect of regulating primary cilia expression on autophagy of chondrocytes. Methods Immunofluorescence staining was used to detect the expression of autophagosomes and primary cilia in chondrocytes, and their localization and expression relationship during cells division cycle. Then, primary cilia were induced by serum-free medium and destroyed by chloral hydrate. Primary cilia incidence, length and autophagy expression were analyzed and their related proteins IFT88, Beclin1 and LC3-Ⅰ/Ⅱ were detected by Western blotting. Results Primary cilia and autophagy could be co-expressed in chondrocytes. The incidence of primary cilia was (62.16±11.74)% and the length was (2.342±0.392) μm, and the relative expression of autophagy was 26.38±5.31 in a single cell. The number, size and distribution of autophagosomes could change with the disassembly and assembly of primary cilia during cell cycle. Serum free medium could induce primary cilia expression, promote prolongation, and up-regulate the autophagy level of chondrocytes and their related proteins IFT88, Beclin1 and LC3-Ⅱ expression. But after disrupting cilia structure by chloral hydrate, the incidence and length of primary cilia were reduced, the expression of autophagy was down-regulated, and the expression of IFT88, Beclin1 and LC3-Ⅱ proteins was inhibited. Conclusion There exist a localization and expression correlation between primary cilia and autophagy in chondrocytes, and regulation primary cilia expression could influence the level of autophagy in chondrocytes. |
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