Modulation of cultured corneal keratocyte phenotype by growth factors/cytokines control in vitro contractility and extracellular matrix contraction
Modulation of cultured corneal keratocyte phenotype by growth factors/cytokines control in vitro contractility and extracellular matrix contraction
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DOI：
10.1016/s0014-4835(03)00188-x
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发表时间：
2003-11-01
影响因子：
3.4
通讯作者：
Jin, HC
中科院分区：
文献类型：
--
作者：
Jester, JV;Jin, HC
本研究的目的是评价特定角膜细胞表型（角膜细胞、成纤维细胞、肌成纤维细胞）的细胞收缩性和收缩细胞外基质的能力。兔角膜基质细胞表型通过暴露于最佳增殖剂量的IGF-I IL-1 α、FGF 2来调节。PDGF-AB和TGF β（1）。然后通过免疫细胞化学、蛋白质印迹、胶原凝胶收缩和LPA刺激来评估细胞，以分别测量：（1）粘着斑（FA）、纤连蛋白（FN）和f-肌动蛋白组装;（2）α-平滑肌肌动蛋白（α-SMA）的表达;（3）收缩细胞外基质的能力和（4）确定收缩能力。未处理的角膜基质细胞没有收缩胶原基质的能力。IGF-I和IL-1 α增加细胞增殖（分别为70.2%和74.3%），但不改变角膜细胞表型或收缩基质的能力。FGF 2和PDGF诱导成纤维细胞分化，FA和FN组装以及显著的（p < 0.05）细胞外基质收缩。TGF β 1诱导肌成纤维细胞分化，FA和FN组装显著，α-SMA表达显著增加（p < 0.05）基质收缩。此外，LPA诱导肌动蛋白丝组装在生长因子饥饿的成纤维细胞和肌成纤维细胞，但对培养的角膜细胞表型没有影响。我们首次报道角膜细胞表型是非收缩性的，细胞静止不是一个定义特征。我们进一步确定，环境条件的变化调节角膜细胞表型，导致细胞收缩性和细胞外基质收缩能力方面的生理功能差异。(C)2003 Elsevier Ltd.保留所有权利。
The purpose of this study was to evaluate specific keratocyte phenotypes (keratocyte, fibroblast, myofibroblast) for cell contractility and ability to contract extracellular matrix. Rabbit keratocyte phenotype was modulated by exposure to optimal proliferative doses of IGF-I IL-1alpha, FGF2. PDGF-AB, and TGFbeta(1). Cells were then evaluated by immunocytochemistry, western blot, collagen gel contraction and LPA stimulation to measure: (1) focal adhesion (FA), fibronectin (FN) and f-actin assembly; (2) expression of alpha-smooth muscle actin (alpha-SMA); (3) ability to contract extracellular matrix and (4) determine contractile ability, respectively. Untreated keratocytes showed no ability to contract collagen matrix. IGF-I and IL-1alpha increased cell proliferation (70.2 and 74.3%, respectively) but did not alter keratocyte phenotype or ability to contract matrix. FGF2 and PDGF induced fibroblast differentiation with FA and FN assembly and significant (p < 0.05) extracellular matrix contraction. TGFbeta(1) induced myofibroblast differentiation with prominent FA and FN assembly, expression of alpha-SMA and significantly greater (p < 0.05) matrix contraction. Addition of LPA induced actin filament assembly in growth factor starved fibroblasts and myofibroblasts but had no effect on the cultured keratocyte phenotype. We report for the first time that the keratocyte phenotype is noncontractile and that cell quiescence is not a defining characteristic. We further establish that changes in environmental conditions modulate the keratocyte phenotype resulting in physiologically functional differences regarding cell contractility and capacity to contract extracellular matrix. (C) 2003 Elsevier Ltd. All rights reserved.