中文摘要：

背景研究表明，转化生长因子β2（TGF-β2）促进Tenon囊成纤维细胞（TFs）的活化，在青光眼滤过术后结膜下纤维化过程中发挥作用，但其作用机制尚未完全明了。赖氨酰氧化酶家族（LOXs）与细胞外基质重塑有关，了解青光眼术后滤过泡的纤维化过程中TGF-β2与LOXs活化的关系对青光眼滤过术后滤过泡瘢痕化的防治具有重要意义。目的观察TGF-β2对体外培养的人眼TFs中LOXs蛋白表达变化的影响。方法将培养的第4～8代人TFs分为正常对照组和不同质量浓度TGF-β2培养组，分别在细胞培养液中添加终质量浓度为2、4、8和16ng／ml TGF-β2继续培养24h，采用Western blot法测定不同质量浓度TGF-β2培养组细胞中LOXs蛋白表达的变化，确定最适TGF-β2刺激质量浓度。用4ng／ml TGF-β2分别处理培养的TFs6、12、24和48h，采用Western blot法测定各时间点细胞中LOXs蛋白表达的变化。采用细胞免疫荧光法分别检测正常对照组和4ng／ml TGF-β2培养组TFs中LOX蛋白的表达及其在细胞中的分布。结果Western blot法检测显示，正常对照组及2、4、8和16ng／ml TGF-β2培养组人TFs中LOX、LOXL1、LOXL2、LOXL3及LOXL4蛋白的表达强度均随TGF-β2质量浓度的升高而增加，各组间细胞中LOX、LOXL1、LOXL2、LOXL3和LOXL4蛋白相对表达量的总体比较，差异均有统计学意义（F=37．338、13．438、31．067、11．767、15．167，均P〈0．01）。细胞中LOX、LOXL1、LOXL2、LOXL3及LOXL4蛋白相对表达量的升高呈TGF-β2剂量依赖性。正常对照组及4ng／ml TGF-β2刺激人TFs后6、12、24和48h组，细胞中LOXL2蛋白的相对表达量分别为0．68±0．07、1．09±0．10、1．32±0．07、1．50±0．06和1．89±0．12，其表达强度呈时间依赖性，总体比较差异有统计学意义（F=82．832，P=0．000）。正常对照组人TFs中LOX蛋白表达强度较弱，主要位于细胞质中，TGF-β2培养组细胞中LOX蛋白的?

英文摘要：

Background Researches showed that transforming growth factor-β2 （TGF-β2 ） promotes the activity of human Tenon capsular fibroblasts （TFs）, which plays a role in the scarring of filtering blebs after antiglaucoma surgery. However,its mechanism is not fully clear. Lysyl oxidases （LOXs） are important extracellular matrix proteases which can catalyze the cross-linking of collagen and elastin. Investigating the impact of TGF-β2 on the expression of LOXs has a great significance for the understanding of the pathogenesis of filtering bleb scarring and its prevention. Objective This study was to investigate the effect of TGF-I32 on the expression of LOXs in cultured human TFs. Methods The TFs at 4- 8 generations were divided into normal control group and different concentrations of TGF-β2 treated-groups, and 100,200,400,800 μl of TGF-β2 with the final concentration of 2,4,8 and 16 ng/ml was added into the medium to treat human TFs respectively for 24 hours. The LOXs in the cells were detected by Western blot to determine the optimal dose of TGF-β2. The 4 ng/ml TGF-β2（200 μl） was used to treat human TFs for 6, 12,24 and 48 hours respectively, and the change of LOXs expression in the cells over time was assayed by Western blot. The expression and distribution of LOX protein in the normal cells and TGF-β2-treated cells was detected by using immunofluorescence technique. This study was approved by Daping Hospital of Third Military Medical University Ethic Commission. The guardians of the patients who offered the specimen knew the purpose of the study and signed informed consent. Results Western blot assay showed that the expressions of LOX, LOXL1, LOXL2,LOXL3 and LOXL4 in the cells were gradually elevated from the normal control group and 2,4,8,16 ng/ml TGF-β2-treated groups, showing significant differences among the groups （ F = 37. 338,13. 438,31. 067, 11. 767, 15. 167 ,all at P〈0.01 ）. The expression of LOXL2 protein in the ceils was 0. 68±0. 07,1.09±0. 10,1.32±0.07,1.50± 0.06 and 1.89