低氧环境下自噬对颏舌肌卫星细胞增殖与凋亡的影响

口腔医学研究 ›› 2015, Vol. 31 ›› Issue (3): 209-212.

低氧环境下自噬对颏舌肌卫星细胞增殖与凋亡的影响

江丽斌¹, 秦文¹, 刘文佳², 邵金陵¹, 郭涛^1*

1. 第四军医大学口腔医院正畸科,军事口腔医学国家重点实验室陕西西安 710032;
2. 第四军医大学口腔医院组织工程研究中心,军事口腔医学国家重点实验室

收稿日期:2014-11-02 出版日期:2015-03-28 发布日期:2016-04-29
通讯作者: 郭涛,Email: freegt@fmmu.edu.cn
作者简介:江丽斌(1988~ ),女,福建莆田人,硕士,主要从事口腔正畸基础与临床研究工作。
基金资助:
国家自然科学基金资助项目(编号:81100058)

Effect of Autophagy on Proliferation Activity and the Apoptosis of Genioglossus Muscle Satellite Cells in Hypoxia Condition.

JIANG Li-Bin, QIN Wen, LIU Wen-Jia, et al.

State Key Laboratory of Military Stomatology, Department of Orthodontics, School of Stomatology, The Fourth Military Medical University, Xi'an 710032

Received:2014-11-02 Online:2015-03-28 Published:2016-04-29

摘要/Abstract

摘要： 目的:探讨低氧环境下自噬对颏舌肌卫星细胞(Genioglossus muscle satellite cells,GG MuSCs)增殖能力与凋亡蛋白表达的影响。方法:体外培养大鼠颏舌肌卫星细胞, MTT法筛选自噬抑制剂氯喹(chloroquine,CQ)无毒剂量并检测细胞增殖能力。实验设常氧组、常氧+25 μmol/L氯喹组、单纯低氧组、低氧+5 μmol/L氯喹组、低氧+15 μmol/L氯喹组、低氧+25 μmol/L氯喹组。Western blot检测自噬标志蛋白LC3、促凋亡蛋白caspase-3及细胞色素c(cytochrome c,Cyt c)的表达水平。结果:1)Western blot结果显示:与常氧组相比,单纯低氧组LC3-Ⅱ/Ⅰ、caspase-3、Cyt c表达均增加;与单纯低氧组相比,低氧+氯喹组中随着给药浓度增加,LC3-Ⅱ/Ⅰ、caspase-3、Cyt c的表达水平呈递增趋势。2)MTT结果显示:与常氧组相比,单纯低氧组细胞增殖能力显著增强;而低氧+氯喹组中随着给药浓度增加,细胞增殖能力受到明显抑制。结论:低氧诱发的自噬促进颏舌肌卫星细胞存活,抑制自噬导致细胞增殖能力下降,凋亡蛋白表达显著增加。这提示自噬可能是颏舌肌卫星细胞抵御低氧损伤的重要机制。

关键词: 自噬, 低氧, 颏舌肌卫星细胞, 阻塞性睡眠呼吸暂停低通气综合症

Abstract: Objective: To explore the effect of autophagy on proliferation activity and expression of apoptotic protein of genioglossus muscle satellite cells (GG MuSCs) in hypoxia condition. Methods: Male rat GG MuSCs were cultured in vitro. The non-toxic dose of chloroquine (CQ) was determined and the proliferation activity of GG MuSCs was detected by MTT assay. The GG MuSCs were divided into six groups: normoxia group, CQ(25μmol/L) plus normoxia group, hypoxia group, CQ(5μmol/L) plus hypoxia group, CQ(15μmol/L) plus hypoxia group and CQ(25μmol/L) plus hypoxia group. Western blot was used to examine the expression of LC3,caspase-3 and cytochrome c(Cyt c) protein. Results: In contrast with normoxia group, the ratio of LC3-Ⅱ/Ⅰ, caspase-3, Cyt c protein expression and the proliferation activity were increased in hypoxia group. After treatment with autophagy inhibitor CQ, the ratio of LC3-Ⅱ/Ⅰprotein expression in CQ plus hypoxia group was higher than hypoxia group, which showed obviously dose-dependent effect. Meanwhile, the proliferation activity of CQ plus hypoxia group significantly decreased in a dose-dependent manner. Conclusion: Hypoxia can induce the autophagy of GG MuSCs and enhance proliferation activity in hypoxia. Inhibition of autophagy can decrease proliferation activity and aggravate the expression of apoptotic protein. This suggests that autophagy may be a vital mechanism which protect the GG MuSCs from hypoxic damage.

Key words: Autophagy Hypoxia , Genioglossus muscle satellite cells, Obstructive sleep apnea hypopnea syndrome

中图分类号:

R783.5

江丽斌, 秦文, 刘文佳, 邵金陵, 郭涛. 低氧环境下自噬对颏舌肌卫星细胞增殖与凋亡的影响[J]. 口腔医学研究, 2015, 31(3): 209-212.

JIANG Li-Bin, QIN Wen, LIU Wen-Jia, et al.. Effect of Autophagy on Proliferation Activity and the Apoptosis of Genioglossus Muscle Satellite Cells in Hypoxia Condition.[J]. Journal of Oral Science Research, 2015, 31(3): 209-212.

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