Yanai Ryoji, Yamada Naoyuki, Kugimiya Naruji, Inui Makoto, Nishida Teruo
Department of Biomolecular Recognition and Ophthalmology, Yamaguchi University School of Medicine, 1-1-1 Minami-Kogushi, Ube City, Yamaguchi 755-8505, Japan.
Invest Ophthalmol Vis Sci. 2002 Jul;43(7):2122-6.
To investigate both the effects of various growth factors on the proliferation of human corneal fibroblasts and the abilities of these factors to protect the cells from apoptosis.
Cultured human corneal fibroblasts were incubated separately with 11 different growth factors whose receptors are expressed by these cells. Cell proliferation was evaluated by measurement of [(3)H]thymidine incorporation. The activation of the protein kinase Akt, which plays an important role in antiapoptotic signaling, was assessed by immunoblot analysis with antibodies specific for a phosphorylated form of the enzyme. Apoptosis was quantitated by the TdT-mediated dUTP-biotin nick-end labeling (TUNEL) assay.
Of the 11 growth factors examined, platelet-derived growth factor, insulin, insulin-like growth factors-1 and -2, and epidermal growth factor, each stimulated the proliferation of corneal fibroblasts, induced the activation of Akt in these cells, and protected them from apoptosis induced by sodium nitroprusside (SNP). Basic fibroblast growth factor, keratinocyte growth factor, nerve growth factor, and hepatocyte growth factor stimulated cell proliferation but did not induce Akt activation or protect the cells from SNP-induced apoptosis. Transforming growth factor-beta1 and -beta2 did not stimulate proliferation and had no effect on Akt activity or on SNP-induced apoptosis.
In terms of their modulatory effects on the proliferation and apoptosis of human corneal fibroblasts, the 11 growth factors examined can be classified into three groups. These growth factors may both contribute to maintenance of the cornea and coordinate the proliferative and apoptotic responses of corneal fibroblasts during corneal wound healing.
研究多种生长因子对人角膜成纤维细胞增殖的影响以及这些因子保护细胞免于凋亡的能力。
将培养的人角膜成纤维细胞分别与11种不同的生长因子共同孵育，这些细胞表达这些因子的受体。通过测量[³H]胸腺嘧啶核苷掺入来评估细胞增殖。通过使用针对该酶磷酸化形式的特异性抗体进行免疫印迹分析，评估在抗凋亡信号传导中起重要作用的蛋白激酶Akt的激活情况。通过TdT介导的dUTP生物素缺口末端标记（TUNEL）测定法定量凋亡。
在所检测的11种生长因子中，血小板衍生生长因子、胰岛素、胰岛素样生长因子-1和-2以及表皮生长因子，均刺激角膜成纤维细胞的增殖，诱导这些细胞中Akt的激活，并保护它们免于硝普钠（SNP）诱导的凋亡。碱性成纤维细胞生长因子、角质形成细胞生长因子、神经生长因子和肝细胞生长因子刺激细胞增殖，但未诱导Akt激活或保护细胞免于SNP诱导的凋亡。转化生长因子-β1和-β2不刺激增殖，对Akt活性或SNP诱导的凋亡无影响。
就其对人角膜成纤维细胞增殖和凋亡的调节作用而言，所检测的11种生长因子可分为三组。这些生长因子可能有助于角膜的维持，并在角膜伤口愈合过程中协调角膜成纤维细胞的增殖和凋亡反应。
