5F对人翼状胬肉成纤维细胞增生及Fas表达的调控  

Regulation of 5F on proliferation of human pterygia fibroblasts and expression of Fas in vitro

作　　者：陈毓东[1] 王映芬[2] 梁俊文 

机构地区：[1]暨南大学第一临床学院眼科,广州510630 [2]广东医学院附属医院眼科,湛江524000 [3]南海市平洲镇医院,528000

出　　处：《眼科研究》2006年第6期603-606,共4页Chinese Ophthalmic Research

摘　　要：目的观察中药半边旗二萜类化合物5F(5F)对体外培养的人翼状胬肉成纤维细胞(HPFs)凋亡的影响以及对Fas基因表达的调控,探讨该药对HPFs作用的机制。方法应用荧光染色观察HPFs的形态改变,免疫细胞化学法(ICC)检测5F对凋亡调控基因Fas表达的影响。结果5F质量浓度为32μg/ml时诱导HPFs凋亡并使Fas基因表达上调,质量浓度为8μg/ml时Fas阳性表达率与对照组比较无显著统计学差异,32μg/ml时Fas阳性表达率与对照组呈显著统计学差异(P<0·01)。结论5F对HPFs增生的抑制作用与促凋亡基因Fas表达上调密切相关。Objective Pterygium is characterized by hyperplasy of fibroblasts. We observed the effect of diterpenoid compound 5F of Pteris semipinnate L on apoptosis of cultured human pterygium fibroblasts （HPFs） and expression of Fas to evaluate its mechanism for proliferation inhibition. Methods HPFs from fresh pterygium during surgery were incubated in vitro. Cultured generation 3 -6 HPFs were collected and identified with vimentin. Collecting cells were treated with 32 μg/ml of 5F for 12,24,48 hours,and the morphologic changes of cells were observed using fluorescent stain with Hoechste 33342 under the microscope. At 48 hours after treated by 8, 16,32,64,128 μg/ml of 5F, expression of Fas on HPFs were detected using immunocytochemical method. Fas expression in normal lymphocyte served as positive control. Results Normal cells displayed a low blue fluorescence. However, in 32 μg/ml of 5F group, apoptosis HPFs showed stronger fluorescence at 24 hours with chromatic agglutination. After 48 hours, necrotic HPFs appeared in photographic images, showing a round shape with low fluorescence. The positive expression of Fas was detected in cell membrane with a blown staining,while negative reaction cells are dyed into blue color. The expression of Fas on HPFs in 8 ttg/ml of 5F group （7.67% ＋ 0. 41% ） was similar to that in control group （5.66% ＋0. 82% ,P 〉0. 05）. In 32 μ/ml and 64 μg/ml of 5F group,positive Fas expression were 24.9% ± 1.71% and 30. 57% ± 1.66% respectively, showing a significant differences in comparison with control groups （ 5.66%± 0. 82% , P 〈 0.01 ）. A decreasing expression rate was found in 128 μg/ml of 5 F group（ 20. 33% ±0. 85 % ）. Conclusion 5 F induces H PFs apoptosis in low concentration and cell necrosis in higher concentration. The inhibitory effect of 5F on HPFs is related to upregulation of expression of Fas.

关 键 词：翼状胬肉 成纤维细胞 5F 凋亡 FAS 

分 类 号：R777.33[医药卫生—眼科]