麦考酚酸抑制翼状胬肉成纤维细胞的作用机制  

The inhibitory effect of mycophenolic acid on pterygium fibroblast

作　　者：程雪娟 张少斌 林林[2] 王继兵 邹会会[3] 

机构地区：[1]潍坊眼科医院潍坊市眼科研究所,261041 [2]滨州医学院流行病学教研室,264000 [3]潍坊医学院眼科学教研室,261061

出　　处：《中华实验眼科杂志》2013年第9期839-844,共6页Chinese Journal Of Experimental Ophthalmology

摘　　要：背景研究表明麦考酚酸（MPA）可下调或抑制与组织增生和炎症相关细胞因子的表达和分泌，进而抑制组织的增生和炎症反应。翼状胬肉是球结膜组织炎性和增生性病变，MPA对翼状胬肉组织的增生是否有抑制作用尚未见报道。目的研究MPA对翼状胬肉组织成纤维细胞（PFBs）活性的影响，并对其相关机制进行探讨。方法翼状胬肉切除术中获取的组织经组织块培养法体外培养PFBs，并采用波形蛋白抗体免疫组织化学法进行鉴定。将0、0．125、0．250、0．500、1．000μmol／LMPA液加入培养孔中，未加入MPA液者作为对照。采用MTT比色法和5一溴脱氧尿嘧啶（BrdU）掺入法观察各组PFBs的生长情况；用Westernblot法检测各组PFBs中核因子-κB（NF—κB）、p65及NF—κBd抑制剂（IκB—a）蛋白的表达。结果培养的细胞为长梭形，呈漩涡状、放射状排列，波形蛋白表达阳性。MTT比色法检测表明，随着MPA浓度的增加，PFBs的增生值（A560）均逐渐下降，5个浓度组间PFBs增生值的总体比较差异有统计学意义（F=42．874，P〈0．01）。随着MPA作用时间的延长，PFBs的增生值（A560）逐渐下降，差异有统计学意义（F=26．038，P〈0．01）。BrdU免疫荧光染色发现，随着MPA浓度的增加，PFBsDNA合成量（A560）逐渐下降，5个组间总体比较差异有统计学意义（F=175．279，P〈O．05），各浓度MPA组PFBsDNA合成量与对照组比较均明显下降，差异均有统计学意义（P〈O．05）。DAPI免疫荧光染色结果表明，各浓度的MPA作用后均未见PFBs的形态学异常。Westernblot法检测显示，作用72h后，MPA组p65表达量为0．886±0．072，明显低于对照组的1．542±0．124，MPA组PFBs细胞质IκB-α的表达量为2．141±0．305，明显高于对照组的1．559±0．267，差异均有统计学意义（P〈0．05）。结论MPA可抑制PFBs的生长，其作用机制可能与其抑制PFBs的Background Studies showed that macophenolie acid （MPA）down-regulates and inhibits the expression and secretion of tissue growth factor and inflammatory factor, and further impacts the proliferation and inflammation process. Pterygium is an inflammatory and proliferative lesion. Whether MPA has an inhibitory effect on pterygium is unclear. Objective This study was to investigate the antifibrotie effects of macophenolic acid on pterygium fibroblasts （PFBs） in vitro and discuss its mechanism. Methods Pterygium tissue was obtained from pterygium patient during the surgery. PFBs were cultured using explants and identified with vimentin immunohistochemisty. 0,0. 125,0. 250,0. 500,1. 000 p, mol/L MPA were added into the culture medium, respectively, and the ceils were cultured in the medium without MPA as the control group. MTT colorimetry was used to find the optimization effective concentration of MPA and evaluate their inhibitory effect on PFBs, and BrdU fluorescence staining was used to assess the growth statue of PFBs. Expressions of nuclear factor-κB（NF-κB） ,p65 and inhibitor of NF-κB-α（IκB-α）in the cells were detected by Western blot. Results The cells was spindle in shape 3 days after cultured and showed the vortex and radial arrangement with the positive response to vimentin. With the increase of MPA,the proliferative value of PFBs （A56o ）showed gradually decline, with a significant difference among the five groups（ F= 42. 874, P〈0.01 ）. In addition, the proliferative value of PFBs （As60 ）significantly lowed as the prolong of MPA active time（ F=26. 038 ,P〈O. 01 ）. BrdU fluorescence staining showed a significant decrease of DNA synthesis of PFBs with the elevation of MPA dose among the five groups（ F= 175. 279 ,P〈0.05 ） , and the A560of PFBs DNA synthesis in different concentrations of MPA groups was lower than that of the control group（ all at P〈0.05 ）. No apoptotic and necrotic cell was found after MPA action by DAPI staining. The expression level of p65 in

关 键 词：翼状胬肉 成纤维细胞 增生 麦考酚酸 P65 核因子-ΚB 

分 类 号：R777.33[医药卫生—眼科]