神经生长因子干预大鼠骨骼肌卫星细胞增殖及α-肌动蛋白的表达  

Nerve growth factor interferes with proliferation and alpha-actin expression of skeletal muscle satellite cells in rats

作　　者：颜南[1] 司晓凤 曾亮[1] 田伟[1] 单广东 熊礼硕 杨炜杰 王正东[1] Yan Nan;Si Xiaofeng;Zeng Liang;Tian Wei;Shan Guangdong;Xiong Lishuo;Yang Weijie;Wang Zhengdong(Shenyang Medical College,Shenyang 110034,Liaoning Province,China)

机构地区：[1]沈阳医学院,辽宁省沈阳市110034

出　　处：《中国组织工程研究》2021年第13期2030-2035,共6页Chinese Journal of Tissue Engineering Research

基　　金：辽宁省科学技术计划项目(2017225060),项目负责人:王正东。

摘　　要：背景:临床研究证实,向神经受损部位注射神经生长因子可改善骨骼肌的运动功能。目的:观察神经生长因子对大鼠原代骨骼肌卫星细胞增殖的影响。方法:采用组织块培养法培养大鼠原代骨骼肌卫星细胞。将第3代骨骼肌卫星细胞分3组培养:对照组加入培养液,低、高浓度组分别加入含10,20 U/mL神经生长因子的培养液。培养2,4,6 d时,倒置相差显微镜与苏木精-伊红染色观察细胞形态,免疫组化染色观察细胞α-肌动蛋白表达;培养1,2,3,4 d后,采用CCK-8法检测细胞增殖活性。结果与结论:①倒置相差显微镜显示,随着培养时间的增加,各组骨骼肌卫星细胞数量逐渐增多,细胞形态由圆形逐渐变为细长的梭形、纺锤形或多角形,并逐渐融合并沿同一个方向排列,并且高浓度组细胞数量多于低浓度组、对照组(P<0.05);②苏木精-伊红染色显示,各组骨骼肌卫星细胞轮廓清晰可见,密集分布,胞核较大且深染,组间无明显差异;③免疫组化染色显示,各组均可见α-肌动蛋白阳性表达,3组间α-肌动蛋白阳性表达吸光度值比较差异无显著性意义(P>0.05);④CCK-8检测显示,低、高浓度组细胞活力高于对照组(P=0.000);⑤结果表明,高、低浓度的神经生长因子均能促进骨骼肌卫星细胞的增殖,但对α-肌动蛋白表达无影响。BACKGROUND:Clinical studies have confirmed that the injection of nerve growth factor into the nerve damage site can improve the motor function of skeletal muscle.OBJECTIVE:To observe the effect of nerve growth factor on the proliferation of primary skeletal muscle satellite cells in rats.METHODS:The original generation of rat skeletal muscle satellite cells were cultivated by the tissue block method,and the third generation of skeletal muscle satellite cells were divided into three groups.The control group was added with culture medium;the low concentration group and the high concentration group were added with culture medium containing 10,20 U/mL nerve growth factors,respectively.At 2,4 and 6 days after culture,the cell morphology was observed by inverted phase contrast microscope and hematoxylin-eosin staining,and the expression ofα-actin was observed by immunohistochemistry.At 1,2,3,and 4 days after culture,CCK-8 assay was used to detect the cell proliferation activity.RESULTS AND CONCLUSION:(1)Inverted phase contrast microscope displayed that with the increase of culture time,the number of skeletal muscle satellite cells in each group increased gradually,and the cell morphology gradually changed from round to fusiform,spindle or polygonal,and gradually fused and arranged along the same direction,and the number of cells in high concentration group was more than that in low concentration group and control group(P<0.05).(2)Hematoxylin-eosin staining showed that the outline of satellite cells in skeletal muscle of each group was clear and densely distributed,and the nucleus was large and deeply stained;there was no significant difference among groups.(3)Immunohistochemical results showed that the expression ofα-actin in skeletal muscle in each group was positive,but there was no statistical difference in the optical density ofα-actin positive particles in each group(P>0.05).(4)The CCK-8 assay results showed that cell viability of low and high concentration groups was higher than that of control group(P=0.000).

关 键 词：骨 骨骼肌 卫星细胞 因子 神经生长因子 细胞培养 肌动蛋白 细胞增殖 

分 类 号：R459.9[医药卫生—治疗学] R319[医药卫生—临床医学]