Department of Ophthalmology, Dongguk University Ilsan Hospital , Goyang , South Korea and.
Curr Eye Res. 2013 Oct;38(10):1072-82. doi: 10.3109/02713683.2013.804093. Epub 2013 Jun 21.
To investigate the effects of Ginkgo biloba extract (GBE) on expression of hypoxia-inducible factor (HIF)-1α and vascular endothelial growth factor (VEGF) in retinal pigment epithelial (RPE) cells under chemical hypoxia.
RPE cells (ARPE-19) were cultured in either untreated media (control group), media treated with 200 μM cobalt chloride (hypoxia group) or media treated with both 200 μM cobalt chloride and 100 μg/ml GBE (hypoxia + GBE group) for various amounts of time. HIF-1α and VEGF expression were compared between groups. HIF-1α and VEGF messenger RNA (mRNA) were quantified using real-time polymerase chain reaction (PCR). HIF-1α of extracted nuclei and VEGF of the media were quantified using enzyme-linked immunosorbent assay (ELISA). The expression of HIF-1α was also assessed with Western blot and immunocytochemistry.
HIF-1α mRNA, VEGF mRNA, HIF-1α and VEGF levels were higher in the hypoxia group compared with the control group; however, real-time PCR revealed decreased expression of HIF-1α mRNA and VEGF mRNA in the hypoxia + GBE group compared with the hypoxia group. In the ELISA, the HIF-1α and VEGF protein concentrations also decreased with GBE treatment. Western blot and immunostaining showed that the intensity of HIF-1α signals in the hypoxia + GBE group was lower than that of the hypoxia group.
GBE reduced HIF-1α and VEGF expression in RPE cells cultured under chemical hypoxia in vitro.
研究银杏叶提取物（GBE）对化学缺氧条件下视网膜色素上皮（RPE）细胞中缺氧诱导因子（HIF）-1α和血管内皮生长因子（VEGF）表达的影响。
将 RPE 细胞（ARPE-19）分别在未经处理的培养基（对照组）、用 200μM 氯化钴处理的培养基（缺氧组）或同时用 200μM 氯化钴和 100μg/ml GBE 处理的培养基（缺氧+GBE 组）中培养不同时间。比较各组间 HIF-1α和 VEGF 的表达。采用实时聚合酶链反应（PCR）定量检测 HIF-1α和 VEGF 信使 RNA（mRNA）。采用酶联免疫吸附试验（ELISA）定量检测提取核中的 HIF-1α和培养基中的 VEGF。还采用 Western blot 和免疫细胞化学法评估 HIF-1α的表达。
与对照组相比，缺氧组 HIF-1α mRNA、VEGF mRNA、HIF-1α 和 VEGF 水平均升高；然而，实时 PCR 显示缺氧+GBE 组 HIF-1α mRNA 和 VEGF mRNA 的表达较缺氧组降低。在 ELISA 中，随着 GBE 处理，HIF-1α 和 VEGF 蛋白浓度也降低。Western blot 和免疫染色显示，缺氧+GBE 组的 HIF-1α 信号强度低于缺氧组。
GBE 降低了体外化学缺氧培养的 RPE 细胞中 HIF-1α 和 VEGF 的表达。
