Objective To study the relationship between fibronectin and primary open-angle glaucoma (POAG) pathogenesis by investigating the effect of fibronectin on the proliferation, adhesion and migration of human trabecular meshwork cells (HTC) in POAG patients. Methods Using HTC that were successfully established from the albuginea oculi in the trabeculectomy of patients, HTC were cultivated for primary culture and subculture. The third passage cells were incubated with different doses of fibronectin (5, 10, 20, 40, 100 μg/ml) cultivated with DMEM/F12. No fibronectin was used in the control group. In order to see the effect of different doses of fibronectin on cell proliferation, adhesion and migration, a Cell Counting Kit-8 and Transwell Kit were used after 24 hours of cultivation. The number of living cells was calculated to detect the optical density value and the data was used to calculate the results, with SPSS 17.0 statistics software. The normal distribution of data between the 2 groups was analyzed with a One-Way ANOVA. SNK test was used to compare the difference between groups. Results POAG trabecular meshwork cells were identified and confirmed with immunocytochemistry and electron microscopy. After 24 hours, a serum -free culture was incubated with different doses of fibronectin (5, 10, 20, 40, 100 μg/ml) cultivated with DMEM/F12. Different doses of fibronectin (5, 10, 20, 40, 100 μg/ml) significantly promote the proliferation of POAG trabecular meshwork cells. However, the proliferation of POAG trabecular meshwork cells with lower doses of fibronectin (5, 10 μg/ml) promote a relatively slow proliferation. There was an upward trend with an increase after the dosage of 10 μg/ml, and proliferation of trabecular meshwork cells reached a peak with higher doses of fibronectin (40 μg/ml). High doses of fibronectin (100 μg/ml) still promote the proliferation of cells, but the proliferation is relatively slow. The difference was significant compared to the control group (P<0.01). Fibronectin (5, 10, 20, 40, 100 μg/ml) promotes adhesion and migration of POAG trabecular meshwork cells, that showed an upward trend. The difference was significant compared to the control group (P<0.05). Conclusion Based on this data, it is possible that fibronectin affects intraocular pressure by affecting the proliferation, adhesion and migration of trabecular meshwork cells POAG patients that control outflow in the trabecular meshwork.%目的 通过研究纤维连接蛋白(FN)对体外培养的原发性开角型青光眼(POAG)患者小梁网细胞增殖、黏附、迁移的影响,探讨FN与POAG发病机制的关系.方法 取临床确诊的POAG患者小梁网切除术中的深层巩膜组织块,进行小梁网细胞体外原代和传代培养,并对其进行免疫细胞化学和电镜鉴定.取第3代小梁网细胞,实验组分别加入浓度为5、10、20、40、100μg/ml的FN(含无血清DMEM/F12培养液)培养,对照组不加FN.培养24 h后,采用CCK-8比色法和Transwell试剂盒检测各组光密度值,分析FN对POAG小梁网细胞增殖、黏附、迁移的影响.采用SPSS 17.0统计软件,组间比较采用One-Way ANOVA分析,两两比较采用SNK检验.结果 经过免疫细胞化学和电镜鉴定后,确定培养的细胞为POAG小梁网细胞.不同浓度FN对小梁网细胞均有影响.FN对POAG小梁网细胞有促增殖作用,在FN为5～10μg/ml浓度范围,小梁网细胞增殖相对缓慢,10μg/ml以后呈现上调趋势,在40μg/ml时达到增殖高峰.100 μg/ml仍促进增殖,但是相对缓慢.各实验组光密度值分别与阴性对照组比较,差异均有统计学意义(P<0.01);各实验组不同浓度组间比较,差异有统计学意义(F=81.778,P<0.05).FN浓度为5、10、20、40、100 μg/ml时,小梁网细胞的增殖率分别为18.6%、54.7%、67.9%、98.7%和121.5%.同样,FN对POAG小梁网细胞有明显的促黏附、迁移作用.小梁网细胞的黏附、迁移能力随FN浓度的增加而增强,基本呈现曲线上升趋势,与阴性对照组比较,差异均有统计学意义(P<0.05).结论 FN能促进POAG患者小梁网细胞增殖、黏附和迁移,其在POAG发病机制中的作用可能为通过影响POAG小梁网细胞的增殖、黏附、迁移功能参与房水流出的凋节过程.
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