CAJ | 학술논문

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。
목적：탐토기아배경중자서작용재인아주막세포중적발생정황。방법：원대배양인아주막성섬유세포，장기수궤분위정상대조조여실험조。정상대조조세포용함10％태우혈청적DMEM 배양기배양；실험조세포용기아유도방법，분별용액이씨평형염용액（EBSS）배양1h、2h、3h、4h、6h ；응용투사전자현미경관찰조간인아주막세포중자서체수량적변화，채용세포면역형광염색화면역인적방법검측인아주막세포중자서상관단백LC3적표체。결과：투사전자현미경결과현시경EBSS기아유도적실험조세포균출현자서체，수량명현다우대조조，차재EBSS유도2h조、3h조、4h조교다。세포면역형광결과표명，경EBSS 기아유도적세포LC3표체량고우대조조，차재EBSS유도2h、3h、4h조상대교고。면역인적검측결과현시，경 EBSS 기아유도적세포 LC3-Ⅱ/LC3-Ⅰ비치명현고우대조조（P＜0．05）。결론：기아유도배경가유도인아주막성섬유세포출현자서작용증다표현。
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<0 . 05) .Conclusion:The starvation condition can induce autophagy of periodontal ligament cells .