摘要
目的 探讨细胞间粘附分子1(ICAM1)在角膜基质细胞中的表达,以及炎症介质对其表达的调节作用。方法 采用细胞培养、免疫细胞化学和流式细胞技术,观察人角膜基质细胞ICAM1的表达及脂多糖(LPS)和干扰素γ(IFNγ)对此表达的影响。结果 体外培养的基质细胞可基础表达一定量的ICAM1,LPS和IFNγ可上调其表达水平至基础表达的14～46倍。结论 角膜炎症反应中,炎症介质促进基质细胞表达ICAM1,与白细胞表面的配体结合导致白细胞的迁移、聚集和局部炎症反应的产生。
ObjectiveTo investigate the expression of intercellular adhesion molecules 1(ICAM 1) and its regulation by inflammatory factors in human corneal keratocytes.MethodsHuman corneal keratocytes were established in 10 d to 30 year old donor eyes.The cubes of corneal stroma were cultured in media consisting of DMEM supplemented with 20% fetal calf serum.The second passage cells were seeded at high density,incubated to subconfluent,and activated with lipopolysaccaride (LPS) and interferon γ(IFN γ).The constitutive expression of ICAM 1 by the third passage human corneal keratocytes was assessed by immunocytochemical staining and flow cytometry analysis.In addition,the regulation of the expression of ICAM 1 by LPS and IFN γ was determined.Results The behavioral characteristics and morphological features of third passage cultured human corneal keratocytes were similar to those in vivo.These cells were suitable for experiments of corneal diseases in vitro.Immunohistochemical staining demonstrated that cultured human corneal keratocytes can constitutively express certain amount of ICAM 1.After exposure 24?h to either LPS or IFN γ,progressive increased significantly in ICAM 1 staining occurred.Flow cytometry analysis determined that ICAM 1 was constitutively expressed by cultured corneal keratocytes.After LPS and IFN γ stimulation for 12 and 48?h respectively,there was a median fold increase in expression of 1 4～4 6.The upregulation of LPS and IFN γ on ICAM 1 by keratocytes was related to the stimulating period and concentration of each factor.Conclusions In corneal inflammation,inflammatory factors can enhance the expression of ICAM 1 in keratocytes.Its binding with the ligand on the surface of leukocytes leads to migration and aggregation of leukocytes in inflammed cornea and thus results in local inflammation.
出处
《眼科研究》
CSCD
2000年第1期4-7,共4页
Chinese Ophthalmic Research
基金
卫生部资助!项目 (项目编号 :94 2 1 97)
关键词
角膜基质细胞
脂多糖
干扰素-Γ
ICAM-炎症
intercellular adhesion molecules 1 human corneal keratocytes lipopolysaccaride interferon γ
