低氧对C2C12细胞分化不同阶段自噬和肌细胞生成素表达的影响

doi:10.13701/j.cnki.kqyxyj.2020.01.017

口腔医学研究 ›› 2020, Vol. 36 ›› Issue (1): 70-74.DOI: 10.13701/j.cnki.kqyxyj.2020.01.017

低氧对C2C12细胞分化不同阶段自噬和肌细胞生成素表达的影响

毛家奇^1,2, 黄蔚², 於丽明^2,3, 赵力如^1,2, 杨冬茹^1*, 卢芸^2,3*

1. 河北医科大学口腔医院口腔内科河北石家庄 050017;
2. 复旦大学附属口腔医院口腔生物医学工程实验室上海 200001;
3. 复旦大学附属口腔医院正畸科上海 200001

收稿日期:2019-07-09 出版日期:2020-01-28 发布日期:2020-01-16
通讯作者: 卢芸,E-mail:luyun_dentist@163.com
杨冬茹,E-mail:yangdr@hebmu.edu.cn
作者简介:毛家奇(1993~),男,四川简阳人,硕士在读,医师,研究方向:正畸与牙周在口腔领域中的相关性及其机制相关研究。
基金资助:
国家自然科学基金 (编号:81600897、81771109)

Effects of Hypoxia on Autophagy and Myogenin Expression in Different Stages of Differentiation of C2C12 Cells.

MAO Jiaqi^1,2, HUANG Wei², YU Liming^2,3, ZHAO Liru^1,2, YANG Dongru^1*, LU Yun^2,3*

1. Department of Endodontics, Stomatological Hospital, Hebei Medical University, Shijiazhuang 050017,China;
2. Oral Biomedical Engineering Laboratory, Shanghai Stomatological Hospital, Fudan University, Shanghai 200001,China;
3. Department of Orthodontics, Shanghai Stomatological Hospital, Fudan University, Shanghai 200001, China.

Received:2019-07-09 Online:2020-01-28 Published:2020-01-16

摘要/Abstract

摘要： 目的:研究低氧对C2C12细胞分化不同阶段自噬相关蛋白Beclin1、微管相关蛋白轻链3(microtubule-associated protein light chain 3,LC3B)和肌细胞生成素(myogenin)表达的影响。方法:C2C12细胞诱导分化1、2、3、4 d,在常氧和低氧状态下观察细胞分化情况。免疫细胞化学染色法检测自噬相关蛋白LC3B和Beclin1的表达。Western blot检测Beclin1、LC3B和myogenin的蛋白表达水平,采用SPSS 22.0软件对数据进行统计学分析。结果:1.与常氧组比较,低氧组分化形成的肌管数量较少,形态细小;2.免疫细胞化学染色结果显示,分化1、2、3、4 d低氧组细胞LC3B和Beclin1荧光强度均低于常氧组;3.Western blot结果显示分化前中期低氧组Beclin1、LC3B和myogenin蛋白表达水平显著低于常氧组(P＜0.05)。随着分化时间的增加,低氧状态下LC3B的表达水平进一步降低,而myogenin表达水平回升。结论:低氧抑制C2C12细胞分化;低氧能降低细胞分化前中期Beclin1、LC3B和myogenin蛋白表达水平。

关键词: C2C12, 自噬, 低氧, 分化

Abstract: Objective: To study the effects of hypoxia on the expression of Beclin1, LC3B and myogenin in different stages of differentiation of C2C12 cells. Methods: C2C12 cells were induced to differentiate for 1, 2, 3 and 4 days. The differentiation of C2C12 cells was observed under normoxia and hypoxia. Immunocytochemistry was used to detect the expression of autophagy-related proteins LC3B and Beclin1. Western blotting was used to detect the protein expression levels of Beclin1, LC3B and myogenin. SPSS22.0 software package was used to analyze the data. Results: The number of myotubes in hypoxia group was less than that in normoxia group, and the morphology of cells was smaller. Immunocytochemistry showed that the fluorescence intensity of LC3B and Beclin1 of hypoxia group at 1, 2, 3 and 4 days of differentiation was lower than that in normoxia group. Western blotting showed that the expression levels of Beclin1, LC3B and myogenin in the early and middle stages of differentiation in hypoxic group were significantly lower than the normoxia group (P<0.05). With the increase of differentiation time, the expression level of LC3B decreased further, while the expression level of myogenin increased. Conclusions: Hypoxia inhibits the differentiation of C2C12 cells; hypoxia can reduce the expression levels of Beclin1, LC3B and myogenin in the early and middle stages of cell differentiation.

Key words: C2C12, autophagy, hypoxia, differentiation

毛家奇, 黄蔚, 於丽明, 赵力如, 杨冬茹, 卢芸. 低氧对C2C12细胞分化不同阶段自噬和肌细胞生成素表达的影响[J]. 口腔医学研究, 2020, 36(1): 70-74.

MAO Jiaqi, HUANG Wei, YU Liming, ZHAO Liru, YANG Dongru, LU Yun. Effects of Hypoxia on Autophagy and Myogenin Expression in Different Stages of Differentiation of C2C12 Cells.[J]. Journal of Oral Science Research, 2020, 36(1): 70-74.

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低氧对C2C12细胞分化不同阶段自噬和肌细胞生成素表达的影响

Effects of Hypoxia on Autophagy and Myogenin Expression in Different Stages of Differentiation of C2C12 Cells.

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