转化生长因子-β1通过肌成纤维细胞的分化促进牛角膜成纤维细胞对胶原凝胶的收缩。

Transforming growth factor-beta 1 promotes contraction of collagen gel by bovine corneal fibroblasts through differentiation of myofibroblasts.

作者信息

Kurosaka H, Kurosaka D, Kato K, Mashima Y, Tanaka Y

机构信息

Department of Ophthalmology, Keio University, School of Medicine, Tokyo, Japan.

出版信息

Invest Ophthalmol Vis Sci. 1998 Apr;39(5):699-704.

PMID:9538875

Abstract

PURPOSE

To determine whether the ability of transforming growth factor-beta (TGF-beta) to influence the contractile activity of corneal fibroblasts depends on their differentiation into myofibroblasts.

METHODS

Bovine corneal fibroblasts were cultured on collagen gel in MED 5 medium (F-12 nutrient mixture supplemented with 5% fetal bovine serum) with or without TGF-beta 1 (0.01-10 ng/ml). To evaluate the corneal fibroblast-derived contraction of collagen gel, the thickness of the gel was measured daily for 6 days. The total number of cells on the gel was counted with a Coulter counter. The detection of alpha-smooth muscle actin (alpha-SMA); a marker for myofibroblasts, on these cells was performed immunocytochemically by using a mouse monoclonal antibody against alpha-SMA. The number of myofibroblasts (alpha-SMA-positive cells) was determined.

RESULTS

The control gels containing bovine corneal fibroblasts that were cultured with the MED 5 medium alone significantly contracted to 72.3 +/- 1.2% of their original thickness after 6 days. TGF-beta 1 increased the contraction of collagen gel mediated by bovine corneal fibroblasts in a dose-dependent manner. Approximately 0.2% of the cells on the control gels cultured with MED 5 medium alone were alpha-SMA positive. TGF-beta 1 significantly increased the expression of alpha-SMA in a dose-dependent manner. There was no significant correlation between the thickness of the collagen gel and the total number of cells. However, there was a significant negative correlation between the thickness of collagen gel and the number of myofibroblasts.

CONCLUSIONS

TGF-beta 1 increased the contractile activity of bovine corneal fibroblasts and their ability to differentiate into myofibroblasts. Because contractile activity was correlated with differentiation, the influence of TGF-beta 1 on corneal fibroblast-induced collagen gel contraction may depend on the promotion of myofibroblast differentiation.

摘要

目的

确定转化生长因子-β（TGF-β）影响角膜成纤维细胞收缩活性的能力是否取决于它们向肌成纤维细胞的分化。

方法

将牛角膜成纤维细胞在MED 5培养基（补充有5%胎牛血清的F-12营养混合物）中培养在胶原凝胶上，添加或不添加TGF-β1（0.01 - 10 ng/ml）。为了评估角膜成纤维细胞诱导的胶原凝胶收缩，每天测量凝胶厚度，持续6天。用库尔特计数器计数凝胶上的细胞总数。通过使用抗α-平滑肌肌动蛋白（α-SMA）的小鼠单克隆抗体，对这些细胞进行免疫细胞化学检测α-平滑肌肌动蛋白，这是肌成纤维细胞的标志物。确定肌成纤维细胞（α-SMA阳性细胞）的数量。

结果

仅用MED 5培养基培养的含有牛角膜成纤维细胞的对照凝胶在6天后显著收缩至其原始厚度的72.3±1.2%。TGF-β1以剂量依赖的方式增加了牛角膜成纤维细胞介导的胶原凝胶收缩。仅用MED 5培养基培养的对照凝胶上约0.2%的细胞为α-SMA阳性。TGF-β1以剂量依赖的方式显著增加了α-SMA的表达。胶原凝胶的厚度与细胞总数之间无显著相关性。然而，胶原凝胶的厚度与肌成纤维细胞的数量之间存在显著的负相关。