来氟米特活性代谢物A77 1726对翼状胬肉成纤维细胞增生的影响  

The effect of active metabolite from leflunomide on the proliferation of human pterygium fibroblasts

作　　者：晏雪莹[1] 张明昌[1] 王凯 郝念[1] 张洁[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院眼科,武汉430022 [2]湖北省云梦县人民医院眼科,432500

出　　处：《眼科研究》2009年第3期174-177,共4页Chinese Ophthalmic Research

摘　　要：目的观察来氟米特活性代谢物丙二酸次氯酰胺(A77 1726)对体外培养的人翼状胬肉成纤维细胞(HPFs)增生的影响,寻找翼状胬肉药物辅助治疗和预防复发的新方法。方法采用组织块法原代培养HPFs,筛选A77 1726药物浓度,采用0～200μmol/L的A77 1726作用与体外培养的第3～7代HPFs,分别于24～96h后观察各梯度浓度A77 1726对HPFs的影响。MTT法检测HPFs的增生活性;免疫细胞化学法测定HPFs中增生细胞核抗原(PCNA)的表达程度。结果在20～200μmol/L浓度作用24～72h,A77 1726可呈浓度依赖和时间依赖性地抑制HPFs增生(P<0.05)。A77 1726浓度在20～200μmol/L时能浓度依赖性地抑制HPFs中PCNA的表达(P<0.05)。结论A77 1726可显著抑制体外培养的HPFs增生活性,且这种抑制作用在一定范围内呈浓度依赖性和时间依赖性。Objective Leflunomide has been utilized in the treatment of various autoimmunopathies in ophthalmology in clinic,and its antifibrosis also has been verified.However,its effect on human pterygium fibroblasts （HPFs） is unclear.Present study was to investigate the effect of A77 1726,an active metabolite from leflunomide,on the proliferation of HPFs in vitro in order to find a novel method for the treatment and prevention of pterygium.MethodsHuman pterygium specimen was collected during pterygium excision.Human pterygium tissue masses were cultured to harvest the HPFs and cultured HPFs were identified using Vimentin antibody,α-SMA antibody and immunochemistry.A77 1726 was added into the DMEM containing 3% fetal bovine serum to treat the 3rd-7th passage of HPFs in the concentration among 20-200 μmol/L.MTT was used to assay the biologic activities under the different doses of A77 1726 at the 24th,48th,72nd,96th hour.The expression of proliferating cell nuclear antigen （PCNA） in each group was detected by immunochemistry.ResultsCultured HPFs showed positive stain for α-SMA and Vimentin antibody.The optical absorbance value（A value） was significantly different among different dosage of A77 1726（Fdosage=17.916,P〈0.05;） and different time group（Ftime=22.551,P〈0.05） and presented a considerable decrease upon dosage and action time of A77 1726（P〈0.05）.The inhibitory rate of A77 1726 on proliferation of HPFs was significantly higher with the increase of dosage and treating time of A77 1726.After treated with 20-200 μmol/L A77 1726,the expression of PCNA in HPFs was obviously influenced by the dosage of A77 1726 （H=80.232,P〈0.05）.ConclusionA77 1726 has a potential inhibition on HPFs proliferation in a dose-and time-dependent manner in a limited circumscription.

关 键 词：来氟米特 翼状胬肉 成纤维细胞 增生 

分 类 号：R777.33[医药卫生—眼科]