Inhibition by All-Trans-Retinoic Acid of Transforming Growth Factor-β-Induced Collagen Gel Contraction Mediated by Human Tenon Fibroblasts
Inhibition by All-Trans-Retinoic Acid of Transforming Growth Factor-β-Induced Collagen Gel Contraction Mediated by Human Tenon Fibroblasts
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DOI：
10.1167/iovs.13-13572
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发表时间：
2014-07-01
影响因子：
4.4
通讯作者：
Sonoda, Koh-Hei
中科院分区：
文献类型：
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作者：
Liu, Yang;Kimura, Kazuhiro;Sonoda, Koh-Hei
目的.过度的伤口收缩会导致结膜瘢痕形成。研究全反式维甲酸（ATRA）对体外培养的人Tenon成纤维细胞（HTFs）收缩性的影响。人Tenon成纤维细胞在I型胶原的3D凝胶中培养，并且在不存在或存在TGF-β、ATRA或各种抑制剂的情况下。通过测量凝胶直径来评价胶原凝胶收缩。通过免疫印迹分析检查各种信号传导分子的磷酸化。通过激光共聚焦显微镜检测肌动蛋白应力纤维和粘着斑的形成。全反式维甲酸以浓度和时间依赖性方式抑制由HTFs介导的TGF-β诱导的胶原凝胶收缩。ATRA可减弱TGF-β诱导的黏着斑激酶（FAK）磷酸化以及应力纤维和黏着斑的形成。全反式维甲酸还抑制TGF-β诱导的促分裂原活化蛋白激酶（MAPK）细胞外信号调节激酶（ERK）、p38和c-Jun NH 2-末端激酶（JNK）以及c-Jun和Smad 2/3的磷酸化。此外，TGF-β诱导的胶原凝胶收缩可被ERK、p38或JNK信号传导抑制剂阻断。全反式维甲酸抑制TGF-β诱导的胶原凝胶收缩介导的HTFs，最有可能通过减弱肌动蛋白应力纤维和粘着斑的形成，以及通过MAPK，c-Jun，和Smads信号。因此，全反式维甲酸可通过减弱Tenon成纤维细胞的收缩性来有效抑制结膜瘢痕形成。
PURPOSE. Excessive wound contraction can lead to scar formation in the conjunctiva. The effects of all-trans-retinoic acid (ATRA) on the contractility of human Tenon fibroblasts (HTFs) cultured in three-dimensional (3D) collagen gels were investigated.METHODS. Human Tenon fibroblasts were cultured in 3D gels of type I collagen and in the absence or presence of TGF-beta, ATRA, or various inhibitors. Collagen gel contraction was evaluated by measurement of gel diameter. Phosphorylation of various signaling molecules was examined by immunoblot analysis. The formation of actin stress fibers and focal adhesions was detected by laser confocal microscopy.RESULTS. All-trans-retinoic acid inhibited TGF-beta-induced collagen gel contraction mediated by HTFs in a concentration-and time-dependent manner. The TGF-beta-induced phosphorylation of focal adhesion kinase (FAK) and formation of stress fibers and focal adhesions in HTFs were attenuated by ATRA. All-trans-retinoic acid also inhibited the TGF-beta-induced phosphorylation of the mitogen-activated protein kinases (MAPKs) extracellular signal-regulated kinase (ERK), p38, and c-Jun NH2-terminal kinase (JNK) as well as that of c-Jun and Smad2/3. Furthermore, TGF-beta-induced collagen gel contraction was blocked by inhibitors of ERK, p38, or JNK signaling.CONCLUSIONS. All-trans-retinoic acid inhibited TGF-beta-induced collagen gel contraction mediated by HTFs, most likely by attenuating the formation of actin stress fibers and focal adhesions as well as signaling by MAPKs, c-Jun, and Smads. All-trans-retinoic acid may therefore prove effective for inhibition of conjunctival scarring through attenuation of the contractility of Tenon fibroblasts.