二氢睾酮对小鼠角膜上皮细胞Mucins表达的影响  

Effects of dihydrotestosterone on Mucins expression in corneal epithelial cells of mice

作　　者：黎黎[1] 高鸽[1] 郑璇[1] 王双梅[1] 康前雁[1] 

机构地区：[1]西安交通大学医学院第一附属医院眼科,陕西省西安市710061

出　　处：《眼科新进展》2014年第8期701-704,709,共5页Recent Advances in Ophthalmology

基　　金：国家自然科学青年基金(编号:81100647);陕西省科技攻关项目(编号:2010K16-04)~~

摘　　要：目的探讨不同浓度二氢睾酮(DHT)对角膜上皮黏蛋白Mucins表达的影响。方法体外原代培养BALB/c小鼠角膜上皮细胞,分为空白对照组以及DHT干预组,DHT干预组给予不同浓度(0.05 g·L-1、0.10 g·L-1、0.20 g·L-1、0.50 g·L-1、1.00 g·L-1)DHT干预24 h;采用RT-PCR方法与免疫荧光组织化学方法分别检测Muc1、Muc4 mRNA以及蛋白水平的表达。结果 Muc1、Muc4 mRNA检测结果显示:空白对照组,0.05 g·L-1、0.10 g·L-1、0.20 g·L-1、0.50 g·L-1DHT干预组Muc1 mRNA相对表达量分别为1.00±0.00、0.15±0.01、1.40±0.09、0.07±0.11、0.06±0.02;Muc4 mRNA相对表达量分别为1.00±0.00、0.30±0.28、1.36±0.05、0.43±0.01、0.45±0.01,0.10 g·L-1DHT干预组Muc1、Muc4 mRNA表达水平较空白对照组明显增加,差异均有统计学意义(均为P<0.05);0.05 g·L-1、0.20 g·L-1、0.50 g·L-1DHT干预组表达均下降,差异均有统计学意义(均为P<0.05)。免疫荧光化学检测结果示,以上各组Muc1 OD值分别为1.86±0.01、1.82±0.01、2.03±0.04、1.88±0.01、1.81±0.02,0.10 g·L-1DHT干预组Muc1表达较空白对照组明显增加,0.05 g·L-1、0.50 g·L-1DHT干预组表达均下调,差异均有统计学意义(均为P<0.05);Muc4 OD值分别为1.86±0.00、1.77±0.01、1.84±0.02、1.92±0.00、1.69±0.05,0.10 g·L-1与0.20 g·L-1DHT干预组Muc4表达较空白对照组明显增加,0.05 g·L-1、0.20 g·L-1、0.50 g·L-1DHT干预组表达均下调,差异均有统计学意义(均为P<0.05)。0.50 g·L-1与1.00 g·L-1DHT可影响细胞体外生长。结论一定浓度DHT(如0.10 g·L-1)可上调小鼠角膜上皮细胞Muc1与Muc4的表达水平;而高浓度(0.50 g·L-1与1.00 g·L-1)DHT对角膜上皮细胞存在毒性作用。Objective To study the effects of dihydrotestosterone（ DHT） on Mucins expression in corneal epithelial cells of mice in vitro. Methods Corneal epithelial cells of BALB /c mice were cultured in vitro. Cells were divided into blank control group and DHT-treated group,the cells in DHT-treated group were treated by different concentrations of DHT（ 0. 05 g·L- 1,0.10 g·L- 1,0. 20 g·L- 1,0. 50 g·L- 1,1. 00 g·L- 1） for 24 hours. Expressions of Mucin1 and Mucin4 mRNA and protein were measured by RT-PCR and fluorescence immunohistochemistry,respectively. Results The relative expressive levels of Muc1 mRNA in blank control group, and0. 05 g·L- 1,0. 10 g·L- 1,0. 20 g·L- 1,0. 50 g·L- 1,1. 00 g·L- 1 DHT-treated group were1. 00 ± 0. 00,0. 15 ± 0. 01,1. 4 ± 0. 09,0. 07 ± 0. 11,0. 06 ± 0. 02, respectively; MUC4 mRNA relative levels were 1. 00 ± 0. 00,0. 30 ± 0. 28,1. 36 ± 0. 05,0. 43 ± 0. 01,0. 45 ± 0. 01, respectively.Compared with blank control group,Muc1 and MUC4 mRNA relative levels in 0. 10 g·L- 1 DHT-treated group increased significantly（ all P ＆lt; 0. 05）,but which in 0. 05 g·L- 1,0. 20g·L- 1,0. 50 g·L- 1,1. 00 g·L- 1 DHT-treated group all decreased significantly（ all P ＆lt; 0.05）. IOD values of Muc1 in above groups were 1. 86 ± 0. 01,1. 82 ± 0. 01,2. 03 ± 0. 04,1. 88 ±0. 01,1. 81 ± 0. 02, respectively. Compared with blank control group, IOD values of Muc1 in 0.10 g·L- 1 DHT-treated group increased significantly（ P ＆lt; 0. 05）,but which in 0. 05 g·L- 1,0.50 g·L- 1 DHT-treated group all decreased significantly（ all P ＆lt; 0. 05）; IOD values of Muc4 in above groups were 1. 86 ± 0. 00,1. 77 ± 0. 01,1. 84 ± 0. 02,1. 92 ± 0. 00,1. 69 ± 0. 05, respectively.Compared with blank control group, IOD values of Muc4 in 0. 10 g·L- 1 and 0. 20 g·L- 1 DHT-treated groups increased significantly（ all P ＆lt; 0. 05）,but which in 0. 05 g·L- 1 and0. 50 g·L- 1 DHT-treated group all decreased significantly（ all P ＆lt; 0. 05）. Conclusion Certain concentration of

关 键 词：雄激素 二氢睾酮 黏蛋白 角膜上皮细胞 

分 类 号：R57[医药卫生—消化系统]