摘要
目的:观察鼠神经生长因子(NGF)对大鼠肾上腺髓质嗜铬细胞瘤PC12细胞活性及形态的影响。方法:体外培养PC12细胞,将其分为阴性对照组和诱导分化组。诱导分化组使用浓度为50 ng/ml NGF,阴性对照组加入等量的磷酸盐缓冲液。采用CCK-8实验检测细胞活性。制作细胞爬片后,于显微镜下直接观察活细胞形态。HE染色后,采用Image J软件对图片进行分析。经神经元特异性烯醇化酶(NSE)抗体荧光染色后,于荧光显微镜下观察细胞染色情况。结果:CCK-8实验显示,诱导分化组PC12细胞活性高于阴性对照组(P<0.001)。经Image J软件分析,诱导分化组PC12细胞贴壁面积大于阴性对照组,突起数量多于阴性对照组,轴突长度长于阴性对照组(均P<0.001)。PC12细胞诱导分化后的类神经元样细胞经NSE抗体染色后,在荧光显微镜下可见胞体及轴突均被染色。结论:鼠NGF诱导后PC12细胞活性增加,细胞贴壁面积增加,细胞轴突增多、增长,为后续研究提供了基础。
Objective:To observe the effects of rat nerve growth factor(NGF)on the activity and morphology of rat adrenal medulla pheochromocytoma PC12 cells.Methods:PC12 cells were cultured in vitro and divided into negative control group and induced differentiation group.The induced differentiation group was treated with 50 ng/ml NGF,and the negative control group was treated with the same amount of phosphate buffer.CCK-8 assay was used to detect cell activity.After making cell climbing slices,the morphology of living cells was observed directly under the microscope.After HE staining,the images were analyzed by Image J software.After fluorescence staining with neuron specific enolase(NSE)antibody,the staining was observed under fluorescence microscope.Results:CCK-8 assay showed that the activity of PC12 cells in the induced differentiation group was higher than that in the negative control group(P<0.001).The adherent area of PC12 cells in the induced differentiation group was larger than that in the negative control group,and the number of neurites was more than that in the negative control group,and the axon length was longer than that in the negative control group(all P<0.001)。Under fluorescence microscope,the cell bodies and axons of the neuronal like cells induced by PC12 cells were stained strongly after NSE antibody staining.Conclusion:After rat NGF induction,the cell activity and adherent area of PC12 cells increase,the number and length of cell axons increase,which provides a basis for follow-up research.
作者
叶劲涛
夏晨晨
杨瑞通
李锋涛
程斌
YE Jintao;XIA Chenchen;YANG Ruitong;LI Fengtao;CHENG Bin(Department of Orthopaedics,the Second Affiliated Hospital of Xi’an Jiaotong University,Xi’an 710004,China)
出处
《陕西医学杂志》
CAS
2022年第3期284-288,共5页
Shaanxi Medical Journal
基金
陕西省重点研发计划项目(2020SF-074)。
作者简介
通讯作者:程斌,E-mail:drcheng2006@sina.com。
