成肌细胞体外培养-力学刺激模型与周期性张应力的影响  

Effects of cyclic stretch on in vitro culture-tensile stimulation models of myoblasts

作　　者：张强[1] 王洪玲[2] 丁弦[1] 夏晨蕾 刘丽娟[3] 王爽玉[1] 李建平[1] 贺苗[1] 孙文娜[1] 阎潇[1] 刘文[1] 张月[1] 姚如永[4] 袁晓[1] 

机构地区：[1]青岛大学医学院附属青岛市立医院口腔医学中心,山东省青岛市266071 [2]泰安市口腔医院,山东省泰安市271000 [3]潍坊市人民医院口腔科,山东省潍坊市261000 [4]青岛大学医学院附属医院中心实验室,山东省青岛市266003

出　　处：《中国组织工程研究》2014年第5期669-674,共6页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金资助项目(31170891)~~

摘　　要：背景:内质网应激参与多种疾病的发生发展过程,如动脉粥样硬化、糖尿病、阿尔茨海默病,GRP78是内质网应激的标志蛋白,GRP78的表达水平反映内质网应激的程度。目的:研究周期性张应力对L6大鼠成肌细胞GRP78表达的影响,以明确周期性张应力与内质网应激的关系。方法:在成功构建L6大鼠成肌细胞体外培养力学刺激模型的基础上,采用RT-PCR和Western blot法分析周期性张应力对GRP78 mRNA和蛋白表达的影响。加力组分别给予1,6,12,24 h的力学刺激,加载频率为10 cycles/min,拉伸变形幅度为15%。对照组与实验组在同时种板,不给予力刺激。结果与结论:GRP78 mRNA随着加力时间的延长呈一致上升趋势,与对照组相比,差异均有显著性意义(P<0.05);GRP78蛋白在加力1 h后,蛋白表达量开始升高,加力6 h后蛋白表达量显著升高,到24 h达到峰值,与对照组相比,差异均有显著性意义(P<0.05)。由以上结果得出,在一定时间范围内,周期性张应力可引起内质网应激,并且随着时间的延长逐渐增强;持续的应力刺激,可引起严重的内质网应激导致成肌细胞凋亡。BACKGROUND：Endoplasmic reticulum stress participates in the occurrence and development of many diseases, such as atherosclerosis, diabetes, and Alzheimer’s disease. GRP78 is a marker of endoplasmic reticulum stress. The expression of GRP78 reflects the degree of endoplasmic reticulum stress. 〈br〉 OBJECTIVE：To investigate the effect of cyclic stretch on GRP78 expression of L6 rat myoblasts, and to identify the relationship between cyclic stretch and endoplasmic reticulum stress. 〈br〉 METHODS：In vitro culture-tensile stimulation models of myoblasts of L6 rats were established successful y. The expression of GRP78 of myoblasts exposed to cyclic stretch was determined by reverse transcription-PCR and western blot assay. Stretch groups were subjected to 15%surface elongation at a frequency of 10 cycles per＆amp;nbsp;minute, over a period of 1, 6, 12 and 24 hours. cells were simultaneously seeded on a plate in the control and experimental groups with no stimulation. 〈br〉 RESULTS AND CONCLUSION：The expression of GRP78 mRNA was continuously elevated over time after stretched treatment, and significant differences were detected as compared with the control group （P〈0.05）. GRP78 protein expression began to increase at 1 hour after stretched treatment, was significantly increased at 6 hours, peaked at 24 hours, and significant differences were visible as compared with the control group （P〈0.05）. In conclusion, cyclic stretch induced the occurrence of endoplasmic reticulum stress, which was enhanced with prolonged time. However, prolonged stretch caused severe endoplasmic reticulum stress and leaded to apoptosis of myoblasts.

关 键 词：实验动物 组织构建 GRP78 周期性张应力 内质网应激 L6 成肌细胞 功能矫形 国家自然科学基金 

分 类 号：R318[医药卫生—生物医学工程]