摘要
目的探讨小分子化合物J2对体外培养的大鼠角膜上皮细胞膜表面超微结构的影响。方法实验研究。取SD大鼠原代培养的角膜上皮细胞进行研究。采用密度梯度离心分离法获取大鼠单个核细胞(MNC),将MNC加入角膜上皮细胞共培养,再用小分子化合物J2处理作为实验组,未进行小分子化合物J2处理的MNC及角膜上皮细胞作为对照组,空白对照组仅为角膜上皮细胞。应用流式细胞术检测角膜上皮细胞CD80的表达。采用原子力显微镜(AFM)观测角膜上皮细胞膜表面形态学参数,包括平均粗糙度(Ra)、均方根粗糙度(Rq)、平均低谷高度(Rvm)及波峰至波谷的粗糙度(Rt)。多个样本均数间的总体比较采用单因素方差分析,组间的多重比较采用LSD-t检验。结果实验组、对照组及空白对照组角膜上皮细胞CD80的表达量分别为1.944±0.237、3.128±0.175及1.082±0.120,组间比较差异有统计学意义(F=156.31,P<0.01)。AFM观测角膜上皮细胞膜表面超微结构:实验组、对照组及空白对照组Ra值分别为86.75±12.60、120.23±12.11、61.94±10.62,组间差异有统计学意义(F=306.92,P<0.01);3个组Rq值分别为102.53±9.45、138.30±10.13、91.96±7.25,组间差异有统计学意义(F=361.85,P<0.01);3个组Rvm值分别为-42.21±14.22、-67.36±10.89、-32.18±19.01,组间差异有统计学意义(F=72.22,P<0.01);3个组Rt值分别为437.32±15.66、495.32±13.96、339.92±11.22,组间差异有统计学意义(F=1634.26,P<0.01);3个组两两比较,差异均有统计学意义(P<0.05)。结论经MNC活化的SD大鼠角膜上皮细胞呈CD80高表达,在小分子化合物J2作用后CD80表达减少;应用AFM可以观测到SD大鼠角膜上皮细胞膜表面超微结构经MNC活化后更加粗糙,颗粒状突起明显增加,在小分子化合物J2作用后超微结构的改变有所减轻。
Objective To study the ultrastructural features of the corneal epithelium in the corneal allograft of rats with micromolecular compound J2.Methods An experimental study.Primarily cultured corneal epithelial cells of rats were used.Mononuclear cells(MNCs)by density gradient centrifugation were assigned into the experimental group[MNCs(2 ml)+corneal epithelial cells+J2],control group[MNCs(2 ml)+corneal epithelial cells]and blank group(corneal epithelial cells).Quantity of CD80 expression was obtained by flow cytometry after coculture.Amplitude and height images were obtained by tapping mode atomic force microscopy(AFM)with a scan rate of 2 Hz and an integral gain of 0.3 to 0.5.Statistical analysis of Ra,Rq,Rvm and Rt was performed using the single-factor analysis of variance,and P value was calculated.Results There were obvious differences in the ultrastructure measured by AFM among groups.Ra was 86.75±12.60 in the experimental group,120.23±12.11 in the control group,and 61.94±10.62 in the blank group(F=306.92,P<0.01).Rq was 102.53±9.45,138.30±10.13,and 91.96±7.25,respectively,in the three groups(F=361.85,P<0.01).Rvm was-42.21±14.22,-67.36±10.89,and-32.18±19.01,respectively(F=72.22,P<0.01).Rt was 437.32±15.66,495.32±13.96,and 339.92±11.22,respectively(F=1634.26,P<0.01).The one way analysis of variance showed significant differences in these parameters among groups.Conclusions Corneal epithelial cells of rats activated by MNCs had higher CD80 expression,but the expression became decreased with micromolecular compound J2.The ultrastructure of the corneal epithelium became coarser after MNCs activation,and the number of protrusions increased significantly.The ultrastructural changes were alleviated by J2.
作者
宋静
黄一飞
郭惠玲
王大江
Song Jing;Huang Yifei;Guo Huiling;Wang Dajiang(The Ophthalmology Division of Chinese PLA General Hospital,Beijing 100853,China)
出处
《中华眼科杂志》
CAS
CSCD
北大核心
2021年第8期608-613,共6页
Chinese Journal of Ophthalmology
基金
国家自然科学基金(30973245,30471855)。
