摘要
目的 研究曲伏前列素对地塞米松诱导的人眼小梁细胞肌动蛋白细胞骨架及β黏蛋白表达的影响.方法 对照实验研究.人眼小梁细胞体外培养、传代.实验分为4组,第1组为空白对照组,第2组为地塞米松(1 ×10-6mol/L)处理组,第3组为地塞米松(1×10-6mol/L)+曲伏前列素(1×10-6mol/L)处理组,第4组为曲伏前列素处理组.培养的人眼小梁细胞于相差显微镜下观察其形态并拍照,于荧光显微镜下观察细胞骨架结构及β黏蛋白变化,用免疫印迹法半定量检测β黏蛋白的表达水平.组间β黏蛋白表达水平比较采用方差分析,进一步两两比较采用q检验.结果 培养的人眼小梁细胞经地塞米松诱导后微丝重组杂乱,形成交叉连接肌动蛋白网结构,β黏蛋白染色明显加强.地塞米松联合曲伏前列素处理组可部分逆转地塞米松诱导的微丝重组和交叉连接肌动蛋白网结构的形成,使β黏蛋白染色明显减弱.空白组、地塞米松组、地塞米松联合曲伏前列素组及曲伏前列素组β黏蛋白表达相对灰度值分别为0.84±0.03、1.65±0.05、1.21±0.05及0.65±0.04,组间β黏蛋白表达差异有统计学意义(F=143.07,P<0.05).进一步两两比较,除空白组与曲伏前列素组β黏蛋白表达差异无统计学意义(q=2.84,P>0.05)外,空白组与地塞米松组(q=15.32)、空白组与地塞米松联合曲伏前列素组(q=11.40)、地塞米松组与地塞米松联合曲伏前列素组(q=9.38)、地塞米松组与曲伏前列素组(q=16.55)、地塞米松联合曲伏前列素组与曲伏前列素组(q=14.31)比较,β黏蛋白表达差异均有统计学意义(P<0.05).结论 曲伏前列素可以部分逆转由地塞米松诱导的人眼小梁细胞骨架及β黏蛋白的改变,这可能是曲伏前列素通过调节小梁网房水流出途径并降低眼压的机制之一.
Objective To investigate the effect of travoprost on changes of actin cytoskeletal and β-catenin protein in the cultured human trabecular meshwork (HTM) cells treated with dexamethasone (DEX). Methods It was a control experiment study. The HTM cells were cultured in vitro and divided into control group, DEX (1 × 10-6mol/L) group, travoprost (1 × 10-6mol/L) group, and DEX (1 ×10-6mol/L) plus travoprost (1 × 10-6mol/L) group. F-actin in the HTM cells was detected by FITC-phallodin and observed under a fluorescence microscope. The expression of β-catenin was determined by immunofluorescence and western-blot. Statistical analysis was performed using SPSS13.0 software. The difference of β-catenin expression among groups was analyzed through variance analysis and, further by q test. Results The cultured HTM cells were identified by immunohistochemistry. A reorganization of actin cytoskeletal and a formation of cross linked actin networks (CLANs) were seen in the HTM cells treated with DEX, which were partially reversed by the treatment with DEX plus travoprost. An increase of the expression of β-catenin was discovered in the HTM cells treated with DEX, which was also partially reversed by the treatment with DEX plus travoprost. The amount of β-catenin protein in untreated control group, DEX group, DEX plus travoprost group and travoprost group were 0. 84 ± 0. 03,1.65 ± 0. 05, 1.21 ± 0. 05, and 0. 65 ±0. 04, respectively. Expression of β-catenin was significantly ( F = 143.07, P < 0. 05 ) different when compared untreated control group with DEX group ( q = 15. 32 ,P <0. 05 ), untreated control group with DEX plus travoprost group (q = 11.40,P<0. 05), DEX group with DEX plus travoprost group (q =9. 38,P < 0. 05 ), DEX group with travoprost group ( q = 16. 55, P < 0. 05 ), and DEX plus travoprost group with travoprost group (q = 14. 31 ,P < 0. 05 ). No difference was found in untreated control group and travoprost group(q = 2. 84, P > 0. 05
出处
《中华眼科杂志》
CAS
CSCD
北大核心
2011年第4期336-341,共6页
Chinese Journal of Ophthalmology
关键词
氯前列醇
小梁网
地塞米松
细胞骨架
Β连环素
Cloprostenol
Trabecular meshwork
Dexamethasone
Cytoskeleton
Beta catenin