Type I Collagen Accelerates the Spreading of Lens Epithelial Cells through the Expression and Activation of Matrix Metalloproteinases
Type I Collagen Accelerates the Spreading of Lens Epithelial Cells through the Expression and Activation of Matrix Metalloproteinases
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DOI：
10.3109/02713683.2013.853194
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发表时间：
2014-04
影响因子：
2
通讯作者：
A. Shimada;Yoshiki Miyata;H. Kosano
中科院分区：
文献类型：
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作者：
A. Shimada;Yoshiki Miyata;H. Kosano
摘要目的：基质金属蛋白酶（MMP）参与后囊膜混浊（PCO），但促进MMP表达的机制尚未确定。在这项研究中，我们调查是否I型胶原，这是只检测到在老年人或白内障透镜胶囊，影响基质金属蛋白酶的表达和激活的原代培养鸡透镜上皮细胞（LEC）。材料与方法：从鸡胚中分离鸡LEC，并在含有10%胎牛血清（FBS）的Dulbecco改良Eagle培养基中在I型胶原包被的培养皿上培养。使用明胶酶谱法检测分泌的MMPs的活性，并将细胞铺展确定为随机分布的细胞的平均面积。对于一些实验，LEC在广谱MMP抑制剂GM 6001的存在下培养。将在未涂覆的培养皿上培养的LEC用作对照。为了检查MMP在细胞迁移中的参与，在MMP抑制剂存在下进行伤口愈合测定。结果：鸡晶状体上皮细胞组成性表达MMP-2前体。当LEC在I型胶原包被的培养皿上培养时，它们表达MMP-2的活性形式和MMP-9的前体形式。在人LEC系SRA-01/04中也观察到I型胶原的这种表达和活化，但在人Müller神经胶质细胞系MIO-M1中未观察到。I型胶原蛋白增强细胞铺展，MMP抑制剂抑制细胞铺展。I型胶原还促进α-平滑肌肌动蛋白的表达。此外，LEC迁移抑制MMP抑制剂在伤口愈合试验中以剂量依赖性的方式。结论：I型胶原以LEC特异的方式促进MMPs的表达和活化。这些结果表明，I型胶原蛋白可能在PCO的发展中发挥作用。
Abstract Purpose: Matrix metalloproteinases (MMPs) are involved in posterior capsule opacification (PCO), but the mechanisms that promote MMP expression are yet to be determined. In this study, we investigated whether type I collagen, which is only detected in aged or cataractous lens capsules, affects the expression and activation of MMPs in primary-cultured chicken lens epithelial cells (LECs). Materials and methods: Chicken LECs were isolated from chicken embryos and cultured in Dulbecco’s modified Eagle’s medium containing 10% fetal bovine serum (FBS) on type I collagen-coated dishes. The activity of secreted MMPs was examined using gelatin zymography, and cell spreading was determined as the average area of randomly distributed cells. For some experiments, LECs were cultured in the presence of the broad-spectrum MMP inhibitor, GM6001. LECs cultured on uncoated dishes were used as controls. To examine the involvement of MMP in cell migration, a wound-healing assay was performed in the presence of the MMP inhibitor. Results: Chicken LECs constitutively express the pro-form of MMP-2. When LECs were cultured on type I collagen-coated dishes, they expressed the active form of MMP-2 and the pro-form of MMP-9. This expression and activation by type I collagen was also observed in the human LEC line SRA-01/04, but not the human Müller glial cell line, MIO-M1. Type I collagen enhanced cell spreading, which was suppressed by the MMP inhibitor. Type I collagen also accelerated α-smooth muscle actin expression. In addition, LEC migration was inhibited by the MMP inhibitor in a dose-dependent manner in the wound-healing assay. Conclusion: Type I collagen promotes the expression and activation of MMPs in a LEC-specific manner. These results suggest that type I collagen may play a role in PCO development.