摘要
目的:探究小鼠发育过程中胸腺上皮细胞的动力学变化。方法:取2月龄、6月龄、12月龄雌性昆明小鼠各5只,采用颈椎脱臼法处死小鼠,取胸腺组织,制作石蜡切片,HE染色观察小鼠胸腺皮质和髓质的变化,免疫组化染色检测胸腺上皮细胞的分子标记(角质蛋白5、角质蛋白8)、胸腺上皮细胞成熟有关的自身免疫调节因子(Aire)、胸腺上皮祖细胞的标志物[上皮细胞黏附分子(EpCAM)]、干细胞转录因子[性别决定区Y框蛋白2(Sox2)],免疫印迹法检测FoxN1、EpCAM的蛋白表达,qRT-PCR检测FoxN1、Aire的mRNA的表达。结果:2月龄小鼠胸腺的皮质和髓质结构较为完整,边界清晰;6月龄小鼠胸腺的皮质和髓质结构逐渐融合;12月龄小鼠胸腺的皮质和髓质结构较紊乱,皮髓质交界处不清楚,皮质变薄;随着月龄增加,小鼠胸腺组织的结构完整性逐渐丧失。随着月龄的增加,小鼠胸腺中K5、K8、Aire、EpCAM和Sox2阳性细胞逐渐减少,FoxN1蛋白和mRNA、EpCAM蛋白和Aire mRNA的表达量逐渐降低。结论:小鼠胸腺上皮细胞以及上皮干细胞/祖细胞随发育逐渐减少,提示胸腺正常组织结构逐渐丧失,导致胸腺衰老和免疫功能衰退。
Aim:To investigate the dynamic changes of thymic epithelial cells during mice development.Methods:Kunming female mice at age of 2 months, 6 months, and 12 months were used, 5 in each group, respectively.Mice were sacrificed by cervical dislocation.The thymus tissues were taken and paraffin sections were prepared.Immunohistochemical staining was used to detect the molecular markers(keratin 5 and keratin 8) of thymic epithelial cells, the autoimmune regulatory factor(Aire) related to maturation of thymic epithelial cells, the marker of thymic epithelial progenitor cells [epithelial cell adhesion molecule(EpCAM)],and stem cell transcription factor [sex determination region Y-frame protein 2(Sox2)],respectively.The protein expression of FoxN1 and EpCAM was detected by Western blot, and the mRNA expression of FoxN1 and Aire was detected by qRT-PCR.Results:The structure of cortex and medulla of mice thymus at 2 months was relatively complete with clear boundaries.The cortex and medulla of 6-month-old mice gradually fused.The structure of cortex and medulla in thymus of 12-month-old mice was disordered, with unclear cortex-medulla junction and the thinner cortex.With the increase of months, K5,K8,Aire, EpCAM and Sox2 positive cells in the thymus were gradually decreased,and the expressions of FoxN1 protein and mRNA,EpCAM protein and Aire mRNA gradually decreased. Conclusion: The mice thymic epithelial cells and epithelial stem cell/progenitor cells decrease gradually with development,indicating that the normal tissue structure of the thymus was gradually lost,leading to the aging of the thymus and the decline of immune function.
作者
杨书平
郑仲
刘相邑
豆晓婕
刘国红
柴玉荣
YANG Shuping;ZHENG Zhong;LIU Xiangyi;DOU Xiaojie;LIU Guohong;CHAI Yurong(Department of Histology and Embryology,School of Basic Medical Sciences,Zhengzhou University,Zhengzhou 450001)
出处
《郑州大学学报（医学版）》
CAS
北大核心
2023年第1期55-61,共7页
Journal of Zhengzhou University(Medical Sciences)
基金
河南省高等学校重点科研项目(21A310022)。
关键词
胸腺
胸腺上皮细胞
祖细胞
小鼠
衰老
thymus
thymic epithelial cell
progenitor cell
mouse
aging
