【摘要】 目的：探讨核心蛋白多糖（decorin）对体外培养的兔晶状体上皮细胞（lens epithelial cells, LEC）分泌转化生长因子β1（transforming growth factor β1, TGFβ1）和表达α平滑肌肌动蛋白（αSMA）的影响。方法：体外原代培养兔LEC，当细胞接近融合时转板培养，分别设置对照组和实验组，对照组为不用药培养12d；实验组为培养6d后加入不同浓度的decorin（10，1.0，0.1mg/L 3个浓度组）继续培养6d，用ELISA法检测各组培养液上清中TGFβ1的水平，免疫细胞化学和计算机图像分析统计各组细胞中αSMA的表达水平。结果：实验组TGFβ1水平和αSMA水平均明显低于对照组的水平（117.9±18.1→427.2±41.3, 111.7±26.7→427.2±41, 236.0±28.6→427.2±41.3；15.7±8.7→97.2±26.9，16.2±9.5→97.2±26.9，54.9±29.6→97.2±26.9,P<0.01），decorin表现出明显的抑制作用;αSMA的表达量随着TGFβ1水平的下降而下降，两者高度正相关（r=0.995，P<0.01）。结论：Decorin抑制活性TGFβ1的水平，下调αSMA的表达。
【关键词】 后发性白内障；核心蛋白多糖；晶状体上皮细胞；转化生长因子β1；α平滑肌肌动蛋白
Study to decorin inhibiting transdifferentiation of rabbits lens epithelial cells in vitro
GuiLan Zhang, Ming Huo, HaiJiang Zhang
Department of Ophthalmology, the Central Peoples Hospital, Yichang 443003, Hubei Province,China
Abstract
AIM: To investigate the effects of the decorin on secretion of transforming growth factorβ1 (TGFβ1) and the expression of αsmooth muscle actin (αSMA) in the primarily cultured rabbits LEC in vitro.
METHODS: Cultured rabbits lens epithelial cells primarily in vitro, then turned to plates when they nearly fused. Set up control group and experiment group, the control group was training 12 days without drugs, others were decorin treatment group (three concentration groups of 10, 1.0, 0.1mg/L) for 6 days’ treatment after training 6 days. ELISA used in each group in the culture medium supernatant of TGFβ1 levels, statistics expression of αSMA by immunocytochemistry and computer image analysis.
RESULTS: The level of TGFβ1 and αSMA of three experiment groups was significantly lower than the level of control group (P<0.01).And immunocytochemistry figures showed that experiment groups hypertrophied cells and positive staining in cytoplasm was much lower than control group. It demonstrated decorin had significant inhibition effect. Moreover, αSMA decreased accompanying to the drop of TGFβ1 and the both were highly positive correlated(r=0.995，P<0.01).
CONCLUSION: Because decorin inhibits the activity of TGFβ1 to reduced the expression of αSMA.
KEYWORDS: posterior capsular opacification; decorin; lens epithelial cells; transforming growth factor β1; αsmooth muscle actin
0引言
在后发性白内障(posterior capsular opacification，PCO)形成过程中，多种细胞因子发挥着至关重要的作用，而越来越多的研究表明，转化生长因子β1（transforming growth factor β1,TGFβ1）能在体内外引起白内障样改变[1]。TGFβ1是PCO形成机制的重要分子基础。核心蛋白多糖（decorin）是一种小分子的硫酸软骨素类蛋白多糖，由富含亮氨酸的核心蛋白和一条糖胺聚糖链组成，存在于几乎所有细胞外基质中，通过拮抗活性TGFβ1来调控细胞的迁移，增殖及分化，且decorin特异性强，没有免疫原性，组织相容性好[2]。我们通过原代培养兔晶状体上皮细胞（lens epithelial cells,LEC），观察不同浓度decorin对细胞分泌TGFβ1的影响和LEC异常表达αSMA的影响，探讨decorin抑制TGFβ1和αSMA的作用。
1材料和方法
1.1材料 2月龄的家兔30只，雌雄不限，由三峡大学实验动物中心提供。优级胎牛血清FCS（杭州四季青生物工程材料有限公司），DMEM培养基和胰蛋白酶（美国Gibco公司），Decorin（美国Sigma公司），ELISA试剂盒（上海西唐生物科技有限公司），鼠抗αSMA mAb和SP试剂盒（福州迈新生物技术有限公司）。德国Leica Q550CW图像分析系统等。
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