低氧环境下自噬对颏舌肌卫星细胞增殖与凋亡的影响  

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

作　　者：江丽斌 秦文 刘文佳 邵金陵 郭涛 

机构地区：[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

出　　处：《口腔医学研究》2015年第3期209-212,共4页Journal of Oral Science Research

基　　金：国家自然科学基金资助项目(编号:81100058)

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+25μmol/L氯喹组、单纯低氧组、低氧+5μmol/L氯喹组、低氧+15μmol/L氯喹组、低氧+25μmol/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。Objective： To explore the effect of autophagy on proliferation activity and expression of apoptotic pro- tein 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（25btmol/ L） plus normoxia group, hypoxia group, CQ（5μmol/L） plus hypoxia group, CQ（15μmol/L） plus hypoxia group and CQ（25tlmol/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 / Ⅰ protein 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 aggra- vate the expression of apoptotic protein. This suggests that autophagy may be a vital mechanism which protect the GG MuSCs from hypoxic damage.

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 

分 类 号：R766[医药卫生—耳鼻咽喉科]