【摘要】 目的观察增殖细胞核抗原(PCNA)在新西兰大耳兔角膜上皮的表达,进一步了解角膜缘干细胞(LSCs)的增殖情况。方法健康成年新西兰大耳兔30只,体重2.0～2.5kg,雌雄错配,取双侧眼球,角膜上、下、内、外四个部位常规石蜡切片,采用常规组织学HE染色、免疫组织化学SABC法染色及图像分析进行研究,相关数据做统计学处理。结果HE染色显示角膜缘上皮细胞排列紧密而不规则,层数较角膜中央部多,基底层细胞呈矮柱状或立方形,核大而圆,居中着色较深,基底部可见明显乳头样突起。角膜中央上皮基底层细胞较大,呈柱状。免疫组织化学SABC法显示PCNA阳性细胞,核卵圆而大,多位于角膜上、下方角膜缘上皮基底层,在角膜缘浅层及角膜浅层有少量分布,在角膜中央上皮基底层可见极少PCNA阳性细胞出现。图像分析显示每个高倍镜视野中上、下角膜缘与角膜及内、外角膜缘PCNA阳性细胞数量及灰度具有显著性差异(P<0.05)。结论PCNA在角膜上皮的标记细胞主要是LSCs和短暂扩充细胞,即标记处于迅速增殖中的细胞。PCNA在角膜中央及角膜缘的表达反映了LSCs增殖,分化的过程;同时提示上、下方的角膜缘干细胞分布较多。更多还原

【Abstract】 Objective To observe the expression of PCNA in epithelial cornea of New Zealand big ear rabbit and to understand more proliferation of LSCs. Methods The 30 New Zealand big ear rabbits of that were healthy adult male or female. Their up and down,in and out of cornea of eyes were do with the paraffin sections. We observed the morphological characteristic of epithelium of cornea with Hematoxylin-Eosin staining. The PCNA-positive cells were showed by immunohistochemistry technique (PCNA-staining).The PCNA-positive cells were counted with the BioMias software of image analysis. Results HE staining: Limbal epithelial cells were closely unregulaly aggregated and the layers of the cells were more than the central zone of cornea. The cells of the basal layer were appeared small cubic or cylindrical-shape. The small and spherical nuclei were centrally positioned and stained darker blue colour,uneven base,we can see clearly nipples. The central epithelial cells were large and columnar shape. Inmunohistochemical SABC method: There were PCNA-positive cells in the corneal limbal epithelial inside the bottom of the distribution of more lateral,including a small amount of limbal epithelial distribution in the distribution of central corneal epithelial minimal.Image analysis showed central corneal with corneal next,and outside limbal epithelial PCNA positive cells quantity and grayscale difference was significant (P<0.05). Conclusion PCNA in corneal epithelial markup cell main yes LSCs and ephemeral expansion cells,namely marked at rapidly proliferation medium cells. PCNA in corneal central and limbal expression reflects LSCs proliferation,differentiation process,prompted corneal below limbal LSCs distribution more.更多还原