埃罗替尼对小鼠角膜上皮组织病理学及超微结构的影响

目的 探讨埃罗替尼对小鼠角膜上皮组织引起的组织病理学及超微结构改变的影响.方法 30只雄性6～8周龄BALB/c小鼠,随机分为对照组(12只)和实验组(12只),6只不作处理为空白对照.实验组使用20 μmol·L-1埃罗替尼液滴眼,对照组采用PBS滴眼,每天4次.分别在干预后1d、7d、14 d对各组小鼠进行荧光素钠(fluorescent,FL)染色及评分.干预后14 d取小鼠眼球,光学显微镜及电子显微镜下观察角膜上皮及细胞的结构变化.提取角膜蛋白进行Western Blot检测.结果 干预前,实验组和对照组FL评分比较差异无统计学意义(P＞0.05).在干预后1d、7d、14 d,实验组FL评分较干预前明显升高,差异均有统计学意义(均为P＜0.05),而对照组FL评分与干预前无明显改变,差异均无统计学意义(均为P＞0.05);两组间FL评分在干预后1d相比,差异无统计学意义(P＞0.05);两组间FL评分在干预后7d、14 d相比,差异均有统计学意义(均为P＜0.05).实验组小鼠角膜上皮细胞排列紊乱,层数增加;电镜下见角膜上皮表层细胞形态不规则、脱落,微绒毛减少、消失;表层上皮细胞的桥粒、半桥粒连接数目明显减少.实验组角膜表皮生长因子受体表达明显发生变化,两组表皮生长因子受体比较差异有统计学意义(P＜0.05).结论 埃罗替尼会引起小鼠角膜上皮的组织结构及细胞超微结构损坏.其机制可能是通过抑制表皮生长因子受体的活化来影响角膜上皮的.

Effects of erlotinib on histopathology and ultrastructure of corneal epithelium in mice

Objective To investigate the histopathological and ultrastructural changes of corneal epithelium induced by erlotinib in mice.Methods Totally 30 6-8 weeks old male BALB/c mice were divided into three groups:Control group (n =12),experimental group (n =12),another 6 mice did nothing as the blank control.Experimental group used erlotinib eye drops and control group used PBS in both eyes,four times per day.At 1 day,7 days and 14 days after the intervention,corneal fluorescence staining (FL) was observed by slit lamp and graded.On the fourteenth day after the intervention,the eye balls of mice were taken,and the histopathological and ultrastructural changes of corneal epithelium and epithelial cells were observed by optical microscope and electron microscope,respectively.And protein of cornea was measured by Western Blot.Results Before the intervention,there was no significant difference in FL scores between the experimental group and control group (P ＞ 0.05).At 1 day,7 days and 14 days,FL score of experimental group was significantly higher than the groups of non-intervention,the difference was statistically significant (all P ＜ 0.05).While FL score of control group was not statistically significant before and after intervention (all P ＞ 0.05);Compared between two groups,there were statistical differences at 7 days,14 days in FL score (all P ＜ 0.05).In the experimental group,the histopathological changes of murine corneal epithelial cells had disorderly arrangement,increased layers of cells,and the inflammatory cells.Under electron microscope,the morphology of corneal epithelial surface cells was irregular and partially detached.The number of microvilli,desmosomes and hemidesmosomes were significantly decreased when compared to the control group.The expression of p-EGFR in experimental group was significantly less than that in control group,the difference was statistically significant (P ＜ 0.05).Conclusion Erlotinib can damage the tissue structure of corneal epithelium and ultrastructure of corneal epithelial cells in mice.And the mechanism is probably that erlotinib influence the corneal epithelium by inhibiting the EGFR activation.