电子烟对小鼠角膜上皮和结膜杯状细胞的影响

The Influence of E-Cigarettes on the Structure of the Corneal Epithelium and Goblet Cells in Mice

目的:研究电子烟对小鼠角膜上皮组织结构及结膜杯状细胞的影响。方法:实验研究。18只雄性c57BL小鼠,8周龄,随机分为A、B、C组,每组6只。A组不做任何干预,B组采用0 mg尼古丁电子烟进行干预,C组采用12 mg尼古丁电子烟进行干预。每日2次,每次30 min。干预12周后,行苏木精-伊红染色观察角膜上皮情况,透射电子显微镜下观察小鼠角膜上皮组织超微结构变化,免疫荧光染色观察Mucin 5AC阳性表达的杯状细胞的改变。各组间结果数据采用单因素方差分析和事后分析。结果:干预12周后,A组上皮细胞层数为5.0±0.7,而B、C组上皮细胞层数分别为7.0±0.7和7.0±0.6,A组与B、C组间差异有统计学意义(F=18.50,P<0.001),而B组和C组间差异无统计学意义。与A组相比,B、C组角膜上皮微绒毛明显减少(F=153.50,P<0.001),长度变短(F=29.54,P<0.001),排列紊乱。在结膜穹隆部的上皮中,B组和C组的Mucin 5AC阳性细胞数目比A组明显减少(F=420.10,P<0.001),但是B组和C组之间Mucin 5AC阳性细胞数目差异无统计学意义。结论:电子烟会损伤小鼠角膜上皮的组织结构,减少结膜杯状细胞的数目。

Objective:To investigate the effects of e-cigarettes on the structure of the corneal epithelium and goblet cells in mice.Methods:In this experimental study,a total of 18 male c57BL mice were divided into three groups:Group A(control group,n=6),group B(0 mg nicotine group,n=6),group C(12 mg nicotine group,n=6).Groups B and C were treated with e-cigarettes two times a day,30 minutes/time,for 12 weeks.After 12 weeks,the histological and ultrastructural changes were examined by hematoxylin and eosin(HE)staining and transmission electron microscopy(TEM).In addition,goblet cells in the cornea and conjunctiva were also investigated with immunofluorescent staining of mucin 5AC.Data analysis was performed using one-way ANOVA followed by Bonferroni's multiple comparison test.Results:After 12 weeks,the corneal epithelium was significantly thicker in groups B and C than in group A(F=18.50,P<0.001),but there was no significant change observed between groups B and C.There was a drastic decrease in the microvilli of the corneal epithelial cells in groups B and C that could be observed with TEM(F=153.50,P<0.001).Furthermore,the corneal epithelial microvilli were significantly shortened(F=29.54,P<0.001)and disorganized.For goblet cells,no goblet cells were observed in the corneal epithelium in any of the three groups.However,goblet cells in the conjunctiva were dramatically reduced in groups B and C compared to group A(F=420.10,P<0.001),but there was no significant difference between groups B and C.Conclusions:E-cigarettesc damage the ocular surface structure and reduce the goblet cells of the conjunctiva in mice.