层黏连蛋白和纤维连接蛋白对人晶状体上皮细胞生长特性的影响  

Laminin and fibronectin influence growth characteristics of human lens epithelial cells in vitro

作　　者：王欣玲[1] 张劲松[1] 于韬[2] 

机构地区：[1]中国医科大学附属第四医院眼科,沈阳110005 [2]辽宁省肿瘤医院放射科

出　　处：《中华眼科杂志》2005年第4期340-345,共6页Chinese Journal of Ophthalmology

基　　金：辽宁省教育厅课题基金资助项目(2004c041)

摘　　要：目的　探讨层黏连蛋白(Ln)和纤维连接蛋白(Fn)对人晶状体上皮细胞生长特性的影响。方法　选用28只人晶状体的前囊膜,采用组织块法接种于Ln(Ln组)、Fn(Fn组)及2%牛血清白蛋白(对照组)包被的培养皿中。应用倒置显微镜观察组织块周围晶状体上皮细胞生长的时间和形态、成纤维细胞出现的时间。采用间接免疫荧光法显示连接素43、ΔNp63和整合素β1 的表达。采用逆转录聚合酶链反应(RT- PCR)检测ΔNp63和整合素β1 mRNA的水平。结果　Ln组和Fn组晶状体上皮细胞长出时间早[ (2. 4±0 .2)d]、细胞数量多,且Fn组成纤维细胞的生长时间[ ( 7. 6±0. 4 )d]明显早于Ln组[ (12. 7±0. 5)d] (P<0 .05)。Ln组连接素43表达的阳性率(23 /28)明显高于Fn组(14 /28)和对照组(12 /28) (P<0 .05),Fn组ΔNp63 ( 24 /28 )和整合素β1 ( 25 /28 )表达的阳性率明显高于Ln组和对照组(P<0 .05)。RT- PCR检测结果支持间接免疫荧光法的检测结果。结论　Ln可持久促进原代培养的人晶状体上皮细胞增殖,并具有一定维持正常细胞间、细胞与细胞外基质间稳态平衡的作用;Fn在细胞培养早期具有较强的促进细胞增殖的作用,但同时也具有促进上皮细胞分化和成纤维细胞生长的作用。Objective To investigate the effects of Laminin (Ln) and Fibronectin (Fn) on human lens epithelial cells (hLECs) in vitro. Methods Twenty-eight pieces of lens anterior capsule were cut into scraps and seeded on dishes coated with Ln, Fn or BSA. The morphological characters of the hLECs were detected with an inverse microscope. The time points when epithelial or mesenchymal cells grew out were recorded. The expressions of Cx43,ΔNp63 and integrinβ 1 were demonstrated with indirect immunofluorescent microscopy. Reverse transcriptase-polymerase chain reaction (RT-PCR) was used to detect the mRNA level of ΔNp63 and integrinβ 1. Results The hLECs in the Ln and Fn groups grew out much earlier [(2.4±0.2)d] with more viable cells than the control group, but the emergence of fibroblastic cells in the Fn group [(7.6±0.4)d] were also earlier than that of the Ln group [(12.7±0.5)d] ( P <0.05). The positive ratio of Cx43 in the Ln group (23/28) was the highest ( P <0.05), while those of ΔNp63 (24/28) and integrinβ 1 (25/28) in the Fn group were also the highest among the three groups ( P < 0.05). The analyses of RT-PCR products were in accordance with the immunofluorescent results. Conclusion Ln can promote persistently the proliferation of hLECs primarily cultured in vitro and functions in retaining the homeostasis of cell-cell or cell-extracellular matrix. Fn showed strong effects in the differentiation of hLECs in the early stage of the cell culture.

关 键 词：层黏连蛋白 纤维连接蛋白 晶状体 上皮细胞生长 核酸酶 

分 类 号：R776.1[医药卫生—眼科]