The Expression of Bone Morphogenetic Protein 2 and Matrix Metalloproteinase 2 through Retinoic Acid Receptor Beta Induced by All-Trans Retinoic Acid in Cultured ARPE-19 Cells.
The Expression of Bone Morphogenetic Protein 2 and Matrix Metalloproteinase 2 through Retinoic Acid Receptor Beta Induced by All-Trans Retinoic Acid in Cultured ARPE-19 Cells.
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在培养的 ARPE-19 细胞中，全反式视黄酸诱导骨形态发生蛋白 2 和基质金属蛋白酶 2 通过视黄酸受体 Beta 表达。
DOI：
10.1371/journal.pone.0150831
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发表时间：
2016
期刊：
影响因子：
3.7
通讯作者：
Zeng J
中科院分区：
文献类型：
--
作者：
Gao Z;Huo L;Cui D;Yang X;Zeng J
全反式维甲酸(ATRA)在眼部发育中起重要作用。既往研究发现，维甲酸可通过促进视网膜色素上皮(RPE)细胞分泌次级信号因子而影响巩膜重塑的代谢。本研究旨在观察维甲酸对培养的急性视网膜色素上皮19(ARPE-19)细胞骨形态发生蛋白-2(BMP-2)和基质金属蛋白酶-2(MMP2)分泌的影响，探讨维甲酸的信号转导途径。采用逆转录聚合酶链式反应(RT-−)和蛋白印迹法检测维甲酸(10−，9~10 mol/L)对ARPE-19细胞视黄酸受体(RARs)表达的影响。采用实时定量聚合酶链式反应和酶联免疫吸附试验检测全反式维甲酸浓度在10−9~10−5−/L作用24 h和48 h或10-6mol/L作用6~72 h对ARPE-19细胞的作用。用RARβ的拮抗剂LE135证实RARβ对ARPE-19细胞的激活作用。全反式维甲酸作用于ARPE-19细胞24小时和48小时后，RAR-β的表达水平显著升高。RARβ基因表达水平在10−9~10−5 m ol/L浓度范围内呈剂量依赖关系，以10−6 m ol/L浓度作用最强，而RARα和RARγ表达水平无明显变化。Western印迹结果显示，10-ATRA作用于ARPE-19细胞12~72 h后，RARβ蛋白水平呈时间依赖性增加，24 h和48 h显著升高，RARβ拮抗剂LE135可抑制RARβ的上调和ATRA诱导的ARPE-19细胞分泌。全反式维甲酸诱导ARPE-19细胞RARβ表达上调，刺激其分泌骨形态发生蛋白-2和基质金属蛋白酶-2。
All-trans retinoic acid (ATRA) plays an important role in ocular development. Previous studies found that retinoic acid could influence the metabolism of scleral remodeling by promoting retinal pigment epithelium (RPE) cells to secrete secondary signaling factors. The purpose of this study was to investigate whether retinoic acid affected secretion of bone morphogenetic protein 2 (BMP-2) and matrix metalloproteinase 2 (MMP-2) and to explore the signaling pathway of retinoic acid in cultured acute retinal pigment epithelial 19 (ARPE-19) cells. The effects of ATRA (concentrations from 10−9 to 10−5 mol/l) on the expression of retinoic acid receptors (RARs) in ARPE-19 cells were examined at the mRNA and protein levels using reverse transcription-polymerase chain reaction (RT-PCR) and western blot assay, respectively. The effects of treating ARPE-19 cells with ATRA concentrations ranging from 10−9 to 10−5 mol/l for 24 h and 48 h or with 10-6mol/l ATRA at different times ranging from 6h to 72h were assessed using real-time quantitative PCR (qPCR) and enzyme-linked immunosorbent assay (ELISA). The contribution of RARβ-induced activation of ARPE-19 cells was confirmed using LE135, an antagonist of RARβ. RARβ mRNA levels significantly increased in the ARPE-19 cells treated with ATRA for 24h and 48h. These increases in RARβ mRNA levels were dose dependent (at concentrations of 10−9 to 10−5 mol/l) with a maximum effect observed at 10−6 mol/l. There were no significant changes in the mRNA levels of RARα and RARγ. Western blot assay revealed that RARβ protein levels were increased significantly in a time-dependent manner in ARPE-19 cells treated with 10−6 mol/l ATRA from 12 h to 72 h, with a marked increase observed at 24 h and 48 h. The upregulation of RARβ and the ATRA-induced secretion in ARPE-19 cells could be inhibited by the RARβ antagonist LE135. ATRA induced upregulation of RARβ in ARPE-19 cells and stimulated these cells to secrete BMP-2 and MMP-2.