饥饿诱导环境下人牙周膜成纤维细胞的自噬作用研究

目的:探讨饥饿环境中自噬作用在人牙周膜细胞中的发生情况。方法:原代培养人牙周膜成纤维细胞,将其随机分为正常对照组与实验组。正常对照组细胞用含10%胎牛血清的DMEM培养基培养;实验组细胞用饥饿诱导方法,分别用厄尔氏平衡盐溶液(EBSS)培养1h、2h、3h、4h、6h;应用透射电子显微镜观察组间人牙周膜细胞中自噬体数量的变化,采用细胞免疫荧光染色和免疫印迹方法检测人牙周膜细胞中自噬相关蛋白LC3的表达。结果:透射电子显微镜结果显示经EBSS饥饿诱导的实验组细胞均出现自噬体,数量明显多于对照组,且在EBSS诱导2h组、3h组、4h组较多。细胞免疫荧光结果表明,经EBSS饥饿诱导的细胞LC3表达量高于对照组,且在EBSS诱导2h、3h、4h组相对较高。免疫印迹检测结果显示,经EBSS饥饿诱导的细胞LC3-Ⅱ/LC3-Ⅰ比值明显高于对照组(P<0.05)。结论:饥饿诱导环境可诱导人牙周膜成纤维细胞出现自噬作用增多表现。

Study on the autophagy in human periodontal ligament fibroblasts induced by starvation

Objective ：To investigate the autophagy in periodontal ligament cells induced by starvation . Methods ：Periodontal ligament cells were cultured and divided into control group and experimental group .The cells in control group were cultured in DMEM medium with 10% FBS .The cells in the experimental group were treated with EBSS for 1h ,2h ,3h ,4h and 6h ,respectively .The autophagosomes in periodontal ligament cells were observed under transmission electron microscope .The expression of autophagy related prote LC3 was detected through west-ern blot and immunofluorescence staining .Results ：The autophagosomes and the expression of LC3 in the cells trea-ted with EBSS were more than those in the control cells ,especially in the 2h ,3h and 4h EBSS treated group .The ra-tio of LC3-II/LC3-I in the cells treated with EBSS was significantly higher than that in the control group cells （P＆lt;0 . 05） .Conclusion：The starvation condition can induce autophagy of periodontal ligament cells .