| 孙文娜 贺苗 夏晨蕾 丁弦 张强 王爽玉 王芳 张彩霞 姜文心 阎潇 孙同正 余海波 袁晓.自噬在应力诱导成肌细胞凋亡中的作用初探[J].,2016,16(11):2045-2048 |
| 自噬在应力诱导成肌细胞凋亡中的作用初探 |
| The Role of Autophagy in Cyclic Stretching Induced Myoblast Apoptosis |
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| DOI： |
| 中文关键词: 自噬 凋亡 周期性张应力 内质网应激 成肌细胞 |
| 英文关键词: Autophagy Apoptosis Cyclic stress Endoplasmic reticulumstress Myoblast |
| 基金项目:国家自然科学基金项目（31170891）；青岛市市科技局项目（13-1-3-3-nsh） |
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| 中文摘要: |
| 目的：探讨自噬在周期性张应力介导的成肌细胞凋亡中的作用，以明确应力诱导内质网应激引起自噬与凋亡之间的关系。
方法：在成功构建L6 大鼠体外培养--力学刺激模型的基础上，采用Western Blot 法分析周期性张应力对自噬相关蛋白LC3 蛋白
表达的影响，并通过Annexin V-FITC/PI流式细胞术检测细胞凋亡情况。加力组分别给予1，6，12，24 h的力学刺激(拉伸变形率为
15 %，频率为10 循环/min)，3-MA 组和Rapamycin 组在加力2 h 前分别加入自噬抑制剂3- 甲基腺嘌呤和自噬激活剂雷帕霉素并
且加力24 h，0 h 组与实验组在同时种板但是不给予力刺激。采用SPSS17.0 统计软件对以上数据进行统计分析。结果：成肌细胞中
的LC3II/LC3I 值随加力时间延长呈上升趋势，24 h达最高(P<0.05)；抑制组的细胞凋亡率(18.75± 1.06 %)相对于0 h组(0.726±
0.13 %)和加力24 h组(14.84± 1.14 %)的明显升高(P<0.05）；Rapamycin 组相对于加力24 h 组的细胞凋亡率明显下降(8.88± 1.08
%vs 14.84± 1.14 %)，但是细胞凋亡率仍然高于0 h 组的(8.88± 1.08 %vs 0.726± 0.13 %)。结论：在一定时间范围内，周期性张应力
可诱导成肌细胞发生自噬，并且自噬活性与作用时间成正比；自噬可以降低应力介导的成肌细胞凋亡的活性。 |
| 英文摘要: |
| Objective:To study the function of autophagy in which cyclic stress induced-apoptosis in myoblasts and clarity the relationship
between autophagy and apoptosis under the mechanical stress.Methods:Based on the mechanical stimulation model of myoblasts
were constructed successfully in vitro, to analysis effects of cyclic tensile stress of LC3 protein expression by Western Blot. Flow
cytometric analysis of the stretched cells for Annexin V-FITC/PI staining was performed to detect apoptotic cells. Continuous cyclic
stretching for 1, 2, 6, 12 and 24 hours (15%surface elongation at a frequency of 10 cycles/min), the inhibitor group which 3-methyladenine
(3-MA) and the Rapamycin group was added to myoblasts 2 h before cyclic stretch, and then 24 h stretch was applied. The 0 h group
which without mechanical stimulation and the experimental group were cultured at the same time. Using SPSS17.0 statistical software for
statistical analysis of the above data.ResultsThe ratio of LC3II /LC3I increased with the loading time prolonged and the 24 h reached
the highest (compared with the control group, the differences were significant difference, P<0.05). The ratio of apoptotic cells in the
3-MA group (18.75± 1.06 %) is higher than loading 24 h group (14.84± 1.14 %); the ratio of apoptotic cells in Rapamycin group is lower
than loading 24 h group but higher than 0 h group (8.88± 1.08 % vs 14.84± 1.14 %, 8.88± 1.08 %vs 0.726± 0.13 %).Conclusion:Cyclical stretch could induce autophagy in myoblasts, and autophagy activity was increased with the loading time prolonged and the 24 h
reached the highest. Autophagy can decrease the activity of cyclical stretch-apoptosis. |
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