Effect of extracellular matrix on proliferation and differentiation of porcine lens epithelial cells
Effect of extracellular matrix on proliferation and differentiation of porcine lens epithelial cells
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DOI：
10.1007/s00417-004-1116-3
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发表时间：
2005-07-01
影响因子：
2.7
通讯作者：
Lang, GE
中科院分区：
文献类型：
--
作者：
de Jong-Hesse, Y;Kampmeier, J;Lang, GE
背景：透镜上皮细胞（LECs）的增殖和分化是继发性白内障形成的重要机制。与完整的透镜相比，白内障囊外摘除术后，剩余晶状体上皮细胞周围的细胞外基质（ECM）发生了变化。本研究探讨了不同ECM对培养的猪LEC中细胞增殖和α-平滑肌肌动蛋白（α-SMA）表达（肌成纤维细胞的标志物）的影响。方法：将猪LEC分别在威尔斯孔和盖玻片上培养3天（细胞增殖测定）或4天（α-SMA表达），用层粘连蛋白、纤连蛋白、I型胶原或IV型胶原包被。在未涂布的威尔斯孔或盖玻片上培养的LEC作为对照。通过[H-3]-胸苷掺入DNA来测量抑制反应。用小鼠单克隆抗体免疫细胞化学检测α-SMA，并计算α-SMA阳性细胞的相对数量。使用Student非配对t检验进行统计分析。结果如下：纤维连接蛋白包被可显著促进细胞增殖（10，320.5 +/-6，073计数/分钟; p < 0.0001）（平均值± SD），I型胶原（12，507.3 +/-3，914.2 CPM; p < 0.0001）和IV型胶原（9，591.4 +/-4，088 CPM; p < 0.0001），而用层粘连蛋白包被没有效果（1，760.8 +/- 812.6 CPM; p=0.7271）。在无涂层盖玻片上培养4天的α-SMA阳性LEC的比例为12.2 +/-3.51%。使用纤连蛋白（24.3 +/- 4.56%; p=0.0001）和I型胶原蛋白（21.2 +/- 8.48%; p=0.0142）涂层后，该比例显着增加。层粘连蛋白（9.8 +/- 3.67%; p=0.1682）和IV型胶原（9.0 +/-7.09%; p=0.2491）涂层略微降低了α-SMA表达，但该效应无统计学显著性。结论：纤连蛋白和I型胶原刺激培养的猪晶状体上皮细胞的细胞增殖和α-SMA的表达。由于纤连蛋白和I型胶原蛋白通常不存在于成人透镜中，因此它们在白内障手术后可能引入透镜囊中可能在后囊膜混浊的发展中起关键作用。
Background: Proliferation and differentiation of lens epithelial cells (LECs) are important mechanisms of secondary cataract formation. After extracapsular cataract extraction the extracellular matrix (ECM) around the remaining LECs is altered compared with the intact lens. This study investigated the effects of different ECMs on cell proliferation and alpha-smooth muscle actin (alpha-SMA) expression, a marker for myofibroblasts, in cultured porcine LECs. Methos: Porcine LECs were cultured for 3 days (cell proliferation assay) or 4 days (alpha-SMA expression) on wells and glass cover slips, respectively, coated with laminin, fibronectin, type I collagen or type IV collagen. LECs cultured on uncoated wells or cover slips served as control. Proliferative response was measured by [H-3]-thymidine incorporation into DNA. alpha-SMA was detected immunocytochemically with a mouse monoclonal antibody, and the relative numbers of alpha-SMA-positive cells were calculated. Statistical analysis was performed using Student's unpaired t-test. Results: Cell proliferation was significantly increased by coating with fibronectin (10,320.5 +/- 6,073 counts per minute; p < 0.0001) (mean +/- SD), type I collagen (12,507.3 +/- 3,914.2 CPM; p < 0.0001) and type IV collagen (9,591.4 +/- 4,088 CPM; p < 0.0001) compared with control (1,876.5 +/- 998 CPM), whereas coating with laminin had no effect (1,760.8 +/- 812.6 CPM; p=0.7271). The ratio of alpha-SMA-positive LECs cultured on uncoated cover slips for a period of 4 days was 12.2 +/- 3.51%. This ratio was significantly increased by coating with fibronectin (24.3 +/- 4.56%; p=0.0001) and type I collagen (21.2 +/- 8.48%; p=0.0142). Coating with laminin (9.8 +/- 3.67%; p=0.1682) and type IV collagen (9.0 +/- 7.09 %; p=0.2491) slightly decreased alpha-SMA expression, but this effect was not statistically significant. Conclusions: Fibronectin and type I collagen stimulated both cell proliferation and alpha-SMA expression in cultured porcine LECs. Because fibronectin and type I collagen are not normally present in the adult lens, their possible introduction into the lens capsule after cataract surgery may play a critical role in the development of posterior capsule opacification.