层粘连蛋白和纤维连接蛋白影响人晶状体上皮细胞生长特性的研究

Laminin and fibronectin influence adhesion and proliferation of human lens epithelial cells cultured in vitro

目的 探讨层粘连蛋白 (laminin ,Ln)和纤维连接蛋白 (fibronectin ,Fn)对原代培养的人晶状体上皮细胞 (humanlensepithelialcells ,hLECs)的黏附、增生特性的影响 ,揭示细胞外基质成分与维持晶状体透明性之间的关系。方法 撕取晶状体前囊膜 2 0个 ,分别剪碎 ,分 3组接种于Ln、Fn和牛血清白蛋白包被的培养皿。倒置显微镜观察接种后上皮细胞长出的时间、上皮细胞的形态以及成纤维细胞出现的时间。间接免疫荧光法显示上皮细胞的连接素 4 3(connexin4 3,Cx4 3)、ΔNp6 3和整合素 β1(integrinβ1)的表达。反转录聚合酶链反应 (RT PCR)检测ΔNp6 3、integrinβ1mRNA的水平。结果 与对照组相比 ,Ln和Fn组hLECs的活细胞数量明显增多 ,但Fn组成纤维细胞的生长较Ln组和对照组明显提前。间接免疫荧光显示 ,Ln组Cx4 3表达的阳性率明显高于其他两组 (P <0 .0 5 ) ;Fn组ΔNp6 3和integrinβ1表达的阳性率明显高于其他两组 (P <0 .0 5 )。Ln和Fn组的ΔNp6 3和integrinβ1mRNA水平高于对照组。结论 Fn和Ln有促进原代培养的hLECs黏附和增生的作用 ;同时 ,Fn促进了上皮细胞的分化和成纤维细胞的生长 ,而Ln则有利于保持hLECs的形态和功能 ;这对于维持hLECs的生理状态 ,进而维持晶状体的透明具有重要意义。

Objective To investigate the effects of two extracellular matrix components,laminin (Ln) and fibronectin (Fn) on the human lens epithelial cells (hLECs) primarily cultured in vitro,and to study the relationship between these components and cataractogenesis.Methods Twenty pieces of lens anterior capsule were cut into scraps and seeded on dishes coated with Ln,Fn or 2% bovine serum albumin.The morphological characters of the lens epithelial cells,and the day that epithelial or fibroblast-like cells grew out were detected with inverted microscopy.The expression of connexin43,ΔNp63,integrin β 1 was demonstrated with indirect immunofluorescent microscopy.Reverse transcriptase-polymerase chain reaction (RT-PCR) was used to show the mRNA level of ΔNp63 and integrin β 1.Results Ln and Fn groups show more adhesive cells compared with the control group.But the epithelial cells of Fn group changed into fibroblast-like cells much earlier than those of Ln group.With indirect immunofluorescent microscopy,the positive rate of connexin43 in the Ln group is the highest,and that of ΔNp63 and integrin β 1 in the Fn group is also the highest among the three groups.And more mRNA transcriptions can be detected in the treated group contrasting to the control group.Conclusion Ln and Fn can promote the adhesion and proliferation of hLECs.Fn is also capable of facilitating the differentiation of the epithelial cells and the surge of the fibroblast cells,while Ln helps to maintain the function and structure of epithelium.