[Expression of retinoid acid receptors in human scleral fibroblasts and regulation of growth of fibroblasts by retinoic acid].
[Expression of retinoid acid receptors in human scleral fibroblasts and regulation of growth of fibroblasts by retinoic acid].
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DOI：
10.3760/j.issn:0412-4081.2007.08.017
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发表时间：
2007-08
期刊：
影响因子：
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通讯作者：
D. Yan;Xiangtian Zhou;Xiao-yan Chen;F. Lü;Jiao Wang;D.-N. Hu;J. Qu
中科院分区：
文献类型：
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作者：
D. Yan;Xiangtian Zhou;Xiao-yan Chen;F. Lü;Jiao Wang;D.-N. Hu;J. Qu
目的探讨培养的人巩膜成纤维细胞是否存在维甲酸受体及维甲酸(RA)对细胞生长的调节作用。方法分离3个供体的人巩膜成纤维细胞，在改良的Ealge‘s培养液中培养。本研究使用的是第三代或第五代细胞。逆转录-聚合酶链式反应(RT-PCR)检测成纤维细胞维甲酸受体亚型的表达。培养液中分别加入浓度为100.00.0 1、0.10、1.0 0、10.0 0和100.00 nmol/L的RA。用维甲酸治疗6天后进行细胞计数和统计学分析。每种细胞株都被检测了三次。结果培养的成纤维细胞主要呈双极或纺锤形形态，平均每个分裂时间为2~3d。RT-PCR检测到6种RA受体亚型(RARα、RARβ、RAR Gamma、RXRα、RXRβ和RXR Gamma)的mRNA表达。视黄酸浓度为100.00、0.10、1.00、10.00和63.2 nmol/L时，对巩膜成纤维细胞增殖的抑制率分别为2.2%、17.6%、37.6%、43.8%和63.2%。当RA浓度超过0.10nmol/L时，RA处理组细胞增殖明显低于对照组(P<；0.05)。结论培养的人巩膜成纤维细胞存在6种维甲酸受体亚型，维甲酸对其生长有明显的抑制作用。这些发现表明，在近视进展过程中，巩膜是维甲酸的潜在作用部位。
OBJECTIVE The aim of this study was to identify the presence of retinoid acid receptors in cultured human scleral fibroblasts and the effect of retinoic acid (RA) on the regulation of cell growth. METHODS Human scleral fibroblasts from three donors were isolated and cultured in Dulbecco Modified Ealge's Medium. Cells from the third or fifth generation were used in the present study. Reverse Transcription-Polymerase Chain Reaction (RT-PCR) was used to detect the expression of retinoid acid receptor subtypes in the fibroblasts. RA was added to the culture medium at concentrations of 0.01, 0.10, 1.00, 10.00 and 100.00 nmol/L, separately. Cells were counted and statistically analyzed six days after being treated with RA. Each strain of the cells was examined three times. RESULTS Cultured fibroblasts mainly showed a bipolar or spindle-shaped morphology with an average time of 2 to 3 days for each division. RT-PCR showed mRNA expression of six subtypes of RA receptors (RAR alpha, RAR beta, RAR gamma, RXR alpha, RXR beta, and RXR gamma). The proliferation of scleral fibroblasts was inhibited by 2.2%, 17.6%, 37.6%, 43.8% and 63.2% following the treatment with 0.01, 0.10, 1.00, 10.00 and 100.00 nmol/L of RA, respectively. Cell proliferation in RA-treated samples reduced significantly as compared to the control samples (P < 0.05) when the RA concentration exceeded 0.10 nmol/L. CONCLUSIONS Six retinoid acid receptor subtypes are present in cultured human scleral fibroblasts and the growth of scleral fibroblasts was inhibited significantly by retinoic acid. These findings indicate that the sclera is a potential site of action for the retinoid acid during progression of myopia.