Induction by latanoprost of collagen gel contraction mediated by human tenon fibroblasts: Role of intracellular signaling molecules
Induction by latanoprost of collagen gel contraction mediated by human tenon fibroblasts: Role of intracellular signaling molecules
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DOI：
10.1167/iovs.07-0451
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发表时间：
2008-04-01
影响因子：
4.4
通讯作者：
Nishida, Teruo
中科院分区：
文献类型：
--
作者：
Liu, Yang;Ko, Ji-Ae;Nishida, Teruo
目的.青光眼滤过手术的结果受结膜下伤口愈合的影响。研究抗青光眼药物拉坦前列素对三维胶原凝胶培养的人眼球筋膜成纤维细胞（HTFs）收缩性的影响。将HTF在具有拉坦前列素或细胞内信号传导的各种抑制剂的I型胶原凝胶中培养。通过测量凝胶直径来评价胶原凝胶收缩，并且通过测量由培养上清液的酸-热水解产生的羟脯氨酸的量来确定胶原降解。通过免疫印迹分析评估丝裂原活化蛋白激酶（MAPK）、粘着斑激酶（FAK）和肌球蛋白轻链（MLC）的磷酸化，并通过激光共聚焦显微镜检查肌动蛋白应力纤维的形成。拉坦前列素以浓度和时间依赖性方式刺激HTF介导的胶原凝胶收缩。拉坦前列素对HTFs的胶原降解无影响。拉坦前列素诱导MAPK（ERK、p38和JNK）和FAK的磷酸化，以及HTFs中应激纤维的形成。此外，ERK（PD 98059和ERK抑制剂II）、p38（SB 203580）、JNK（JNK抑制剂II）、Rho相关激酶（Y27632）、磷脂酶C（U 73122）和MLC激酶（ML-7）的抑制剂可降低拉坦前列素诱导的胶原凝胶收缩。拉坦前列素诱导由HTFs介导的胶原凝胶收缩。拉坦前列素的这种作用似乎取决于HTFs中应力纤维的形成和MAPK、FAK、Rho相关激酶、磷脂酶C和MLC激酶的活化。因此，拉坦前列素可能通过影响Tenon成纤维细胞的收缩性来影响结膜下伤口愈合。
PURPOSE. The outcome of glaucoma filtration surgery is affected by subconjunctival wound healing. The effects of the antiglaucoma drug latanoprost on the contractility of human Tenon fibroblasts (HTFs) cultured in a three-dimensional collagen gel were investigated.METHODS. HTFs were cultured in a type I collagen gel with latanoprost or various inhibitors of intracellular signaling. Collagen gel contraction was evaluated by measurement of gel diameter, and collagen degradation was determined by measurement of the amount of hydroxyproline generated by acid-heat hydrolysis of culture supernatants. Phosphorylation of mitogen-activated protein kinases (MAPKs), focal adhesion kinase (FAK), and myosin light chain (MLC) was assessed by immunoblot analysis, and the formation of actin stress fibers was examined by laser confocal microscopy.RESULTS. HTF-mediated collagen gel contraction was stimulated by latanoprost in a concentration- and time-dependent manner. Latanoprost had no effect on collagen degradation by HTFs. Latanoprost induced phosphorylation of MAPKs (ERK, p38, and JNK) and FAK, as well as the formation of stress fibers in HTFs. Furthermore, latanoprost-induced collagen gel contraction was reduced by inhibitors of ERK (PD98059 and ERK inhibitor II), p38 (SB203580), JNK (JNK inhibitor II), Rho-associated kinase (Y27632), phospholipase C (U73122), and MLC kinase (ML-7).CONCLUSIONS. Latanoprost induced collagen gel contraction mediated by HTFs. This action of latanoprost appeared to depend on the formation of stress fibers and the activation of MAPKs, FAK, Rho-associated kinase, phospholipase C, and MLC kinase in HTFs. Latanoprost may therefore influence subconjunctival wound healing by affecting the contractility of Tenon fibroblasts.