Fenofibrate inhibits the expression of VEGFC and VEGFR-3 in retinal pigmental epithelial cells exposed to hypoxia.
Fenofibrate inhibits the expression of VEGFC and VEGFR-3 in retinal pigmental epithelial cells exposed to hypoxia.
复制标题
非诺贝特抑制缺氧视网膜色素上皮细胞VEGFC和VEGFR-3的表达
DOI：
10.3892/etm.2015.2697
复制
发表时间：
2015-10
影响因子：
2.7
通讯作者：
Li Y
中科院分区：
文献类型：
--
作者：
Zhao J;Geng YU;Hua H;Cun B;Chen Q;Xi X;Yang L;Li Y
本研究旨在探讨非诺贝特抑制缺氧条件下的人视网膜色素上皮细胞(RPE细胞)刺激人脐静脉内皮细胞(HUVECs)增殖和迁移的机制。为此，将RPE细胞和HUVECs分为RPE常氧组、RPE+非诺贝特组、RPE缺氧组、RPE缺氧组+非诺贝特组、HUVECs正常培养组和HUVECs+RPE缺氧组。用氯化钴(CoCl2)诱导RPE细胞缺氧。使用超氧阴离子探针来测量超氧阴离子的产生，这是低氧条件的指示。用四甲基偶氮唑盐比色法检测细胞增殖，用酶联免疫吸附试验检测培养上清液中血管内皮生长因子C(VEGFC)和血管内皮生长因子受体3(VEGFR-3)的表达。通过划痕实验检测HUVECs的迁移能力，通过细胞腔形成实验检测HUVECs的血管生成能力。RT-qPCR法检测RPE细胞VEGFC和VEGFR-3mRNA表达水平，Western印迹法检测VEGFR-3mRNA和蛋白表达水平。结果表明，非诺贝特可抑制缺氧诱导的RPE细胞培养上清液中VEGFC和VEGFR-3的表达和释放。将HUVECs培养于缺氧的RPE细胞培养上清液中，可提高HUVECs的存活率和迁移能力，并促进管腔形成，这些作用可被非诺贝特抑制。综上所述，我们的研究结果表明，RPE细胞暴露在低氧环境中可以诱导VEGFC和VEGFR-3的表达，并释放到细胞培养上清液中。RPE细胞在低氧条件培养液中培养后，VEGFC和VEGFR-3的表达增加，并促进HUVEC的增殖、迁移和毛细血管的形成，提示RPE细胞在低氧所致脉络膜新生血管的形成中起重要作用。非诺贝特抑制缺氧RPE细胞中VEGFC和VEGFR-3的表达，从而降低HUVECs形成新血管的能力。
The aim of the present study was to examine the mechanisms through which fenofibrate inhibits the ability of human retinal pigment epithelial cells (RPE cells) exposed to hypoxia to stimulate the proliferation and migration of human umbilical vein endothelial cells (HUVECs). For this purpose, RPE cells and HUVECs were divided into the following groups: RPE-normoxia, RPE + fenofibrate, RPE-hypoxia, RPE hypoxia + fenofibrate; HUVECs normal culture and HUVECs + RPE-hypoxia culture supernatant. RPE cell hypoxia was induced by cobalt(II) chloride (CoCl2). A superoxide anion probe was used to measure the production of superoxide anion, which is indicative of hypoxic conditions. Cell proliferation was assessed by MTT assay, and the expression of vascular endothelial growth factor C (VEGFC) and vascular endothelial growth factor receptor-3 (VEGFR-3) in the RPE cell culture supernatant was measured by enzyme-linked immunosorbent assay (ELISA). The migration ability of the HUVECs was determined by scratch-wound assay, and the angiogenic ability of the HUVECs was examined by measuring cell lumen formation. The mRNA and protein expression levels of VEGFC and VEGFR-3 in the RPE cells were measured by RT-qPCR and western blot analysis, respectively. Our results revealed that fenofibrate inhibited the increase in the expression and release of VEGFC and VEGFR-3 into the RPE cell culture supernatant induced by exposure to hypoxia. The culture of HUVECs in medium supernatant of RPE cells epxosed to hypoxia enhanced the viability and migration ability of the HUVECs and promoted lumen formation; these effects were inhibited by fenofibrate. In conclusion, our data demonstrated that the exposure of RPE cells to hypoxia induced the expression and release of VEGFC and VEGFR-3 into the cell culture supernatant. The culture of HUVECs in conditioned medium from RPE cells exposed to hypoxia increased VEGFC and VEGFR-3 expression, and promoted the proliferation and migration of the HUVECs, as well as capillary tube formation, suggesting that RPE cells play an important role in the formation of choroidal neovascularization resulting from hypoxia. Fenofibrate inhibited the upregulation of VEGFC and VEGFR-3 in the RPE cells exposed to hypoxia, and thus reduced the ability of HUVECs to form new blood vessels.