压力对牛眼小梁细胞诱导型一氧化氮合酶mRNA的表达及蛋白质合成的影响  

Pressure influence on mRNA expression and protein synthesis of inducible nitric oxide synthetase in bovine trabecular meshwork cell

作　　者：薛蔚[1] 杜蜀华[1] 李勇[2] 黄恺[2] 

机构地区：[1]同济医科大学附属,武汉430030 [2]同济医院眼科同济医科大学附属协和医院

出　　处：《中华眼科杂志》2000年第4期295-298,I019,共4页Chinese Journal of Ophthalmology

摘　　要：目的　研究牛眼小梁细胞中诱导型一氧化氮合酶 (induciblenitricoxidesynthase,iNOS)在不同压力下的mRNA表达及蛋白质合成的变化 ,探讨一氧化氮 (nitricoxide,NO)在青光眼发病中的作用。方法　培养新生牛眼小梁细胞 ,并分别给予 2 0、40、6 0及 80mmHg(1mmHg =0 133kPa)的压力 ,以不加压组为对照组 ,用原位杂交和还原型辅酶Ⅱ硫辛酰胺脱氢酶 (nicotinamide adeninedinucleotidephosphate ,NADPH)组织化学染色技术对小梁细胞iNOS的mRNA和蛋白质含量变化进行定性及定量观察。结果　牛眼小梁细胞中iNOSmRNA及蛋白质含量均随压力增高而增高。对照组与压力 2 0mmHg组比较 ,两组iNOSmRNA均为弱表达 ,组间差异无显著性 (t=0 95 0 1,P >0 0 5 ) ;压力 40mmHg和 6 0mmHg组与对照组相比差异有显著性 (t值分别为 0 0 381和 0 0 32 4,P <0 0 5 ) ;压力 80mmHg组与对照组相比差异有非常显著性 (t=0 0 0 0 16 ,P <0 0 1) ;压力 40mmHg组与 6 0mmHg组相比 ,组间差异无显著性 (t=0 112 3,P >0 0 5 ) ,而与 80mmHg组相比差异有非常显著性 (t值分别为 0 0 0 2 9和 0 0 0 45 ,P <0 0 1)。NADPH组织化学染色结果与原位杂交结果基本相同 ,仅显示 40mmHg组与6 0mmHg组的组间差异有显著性 (t=0 0 42 0 ,P <0 0Objective To investigate the inducible nitric oxide synthetase (iNOS) mRNA expression and protein synthesis in bovine trabecular meshwork cell and discuss the possible effects of nitric oxide (NO) in the development of glaucoma Methods 20 mm Hg, 40 mm Hg, 60 mm Hg and 80 mm Hg pressure were respectively added on cultured trabecular cells of new born bovine No pressure group was set as the control The changes of iNOS mRNA and protein in trabecular meshwork cells under different pressures were demonstrated qualitatively and quantitatively by in situ hybridization and nicotinamide adenine dinucleotide phosphate diaphorase (NADPH d) histochemical assay Results The expression of iNOS mRNA and its protein synthesis became higher and higher as the pressure was increased There was weak iNOS mRNA expression in the control and 20 mm Hg groups, and no significant difference between them 40 mm Hg and 60 mm Hg groups had a statistical difference from the control group And 80 mm Hg group had a very significant difference from the control group There was no difference between 40mm Hg and 60 mm Hg groups, but they had significant difference from 80 mm Hg group NADPH d histochemical assay showed almost the same results except that there was significant difference between 40 mm Hg and 60 mm Hg groups Conclusions Pressure can evoke the expression of iNOS mRNA, and the nitric oxide thus produced can be one of the causes of trabecular meshwork destruction, that may induce or aggravate glaucoma

关 键 词：一氧化氮 压力 MRNA 青光眼 小梁细胞 

分 类 号：R770.2[医药卫生—眼科]