二氢睾酮对小鼠角膜上皮细胞Mucins表达的影响

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（０．０５ｇ?Ｌ－１、０．１０ｇ?Ｌ－１、０．２０ｇ?Ｌ－１、０．５０ｇ?Ｌ－１、１．００ｇ?Ｌ－１）ＤＨＴ干预２４ｈ；采用ＲＴ-ＰＣＲ方法与免疫荧光组织化学方法分别检测Ｍｕｃ１、Ｍｕｃ４ｍＲＮＡ以及蛋白水平的表达。结果　Ｍｕｃ１、Ｍｕｃ４ｍＲＮＡ检测结果显示:空白对照组，０．０５ｇ?Ｌ－１、０．１０ｇ?Ｌ－１、０．２０ｇ?Ｌ－１、０．５０ｇ?Ｌ－１ ＤＨＴ干预组Ｍｕｃ１ｍＲＮＡ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；Ｍｕｃ４ｍＲＮＡ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±０．０１，０．１０ｇ?Ｌ－１ＤＨＴ干预组Ｍｕｃ１、Ｍｕｃ４ｍＲＮＡ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）；０．０５ｇ?Ｌ－１、０．２０ｇ?Ｌ－１、０．５０ｇ?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Ｍｕｃ１ＯＤ值分别为１．８６±０．０１、１．８２±０．０１、２．０３±０．０４、１．８８±０．０１、１．８１±０．０２，０．１０ｇ?Ｌ－１ＤＨＴ干预组Ｍｕｃ１表达较空白对照组明显增加，０．０５ｇ?Ｌ－１、０．５０ｇ?Ｌ－１ＤＨＴ干预组表达均下调，差异均有统计学意义（均为Ｐ＜０．０５）；Ｍｕｃ４ＯＤ值分别为１．８６±０．００、１．７７±０．０１、１．８４±０．０２、１．９２±０．００、１．６９±０．０５，０．１０ｇ?Ｌ－１与０．２０ｇ?Ｌ－１ＤＨＴ干预组Ｍｕｃ４表达较空白对照组明显增加，０．０５ｇ?Ｌ－１、０．２０ｇ?Ｌ－１、０．５０ｇ?Ｌ－１ＤＨＴ干预组表达均下调，差异均有统计学意义（均为Ｐ＜０．０５）。０．５０ｇ?Ｌ－１与１．００ｇ?Ｌ－１ＤＨＴ可影响细胞体外生长。结论　一定浓度ＤＨＴ（如０．１０ｇ?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（０．５０ｇ?Ｌ－１与１．００ｇ?Ｌ－１）ＤＨＴ对角膜上皮细胞存在毒性作用。

Effects of dihydrotestosterone on Mucinsexpression in corneal epithelial cells of mice

Objective To study the effects of dihydrotestosterone ( DHT) on Mucins expression rn 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 ’ 1’1 ,0. 20 g ’ 1",0. 50 g ’ L-’, 1.00 g . L-’)for 24 hours. Expressions of Mucinl and Mucin4 mRNA and protein were measured by RT-PCR and fluorescence immunohistochemistry, respectively. Results The relative expressive levels of Mucl mRNA in blank control group , and 0. 05 g . L-l , 0. 10 g . 1-1 ,0. 20 g . 1-’,0. 50 g . 1-1 ,1. 00 g . L-I DHT-treated group were l. 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 l. 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, Mucl and MUC4 mRNA relative levels in 0. 10 g . L -l DHT-treated group increased significantly ( all P < 0. 05 ) ,but which in 0. 05 g . 1’1 ,0. 20 g . 1-1 ,0. 50 g . 1-1 ,1. 00 g . L-1 DHT-treated group all decreased significantly ( all P < 0. 05 ). IOD values of Mucl in above groups were l. 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 Mucl in 0. 10 g . L-I DHT-treated group increased significantly ( P < 0. 05 ) .but which in 0. 05 g . L ’1 ,0. 50 g . L -l DHT-treated group all decreased significantly ( all P < 0. 05 ) ;IOD values of Muc4 in above groups were l. 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-l and 0. 20 g ’ L-l DHT-treated groups increased significantly ( all P < 0. 05 ) ,but which in 0. 05 g . L-l and 0. 50 g . L-l DHT-treated group all decreased significantly ( all P < 0. 05 ) . Conclusion Certain concen-tration of DHT (0. 10 g . L-’) can up-regulate Mucl and Muc4 expressions in corneal epithelial cells of mice in vitro, but high concentration of DHT (0. 50 g . L-l and l. 00g . L-’)may be toxic to cells.