Vasohibin-1 and Retinal Pigment Epithelium
Vasohibin-1 and Retinal Pigment Epithelium
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DOI：
10.1007/978-1-4614-0631-0_40
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发表时间：
2012-01-01
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影响因子：
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通讯作者：
Abe, Toshiaki
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文献类型：
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作者：
Ishikawa, Yumi;Nagai, Nobuhiro;Abe, Toshiaki
血管内皮生长因子(VEGF)是诱导老年性黄斑变性(AMD)患者脉络膜新生血管的主要因素之一。视网膜色素上皮(RPE)是表达血管内皮生长因子的主要来源之一。Vasohibin-1是一种血管内皮生长因子诱导基因，在培养的人内皮细胞中具有抗血管生成的特性。我们观察了血管紧张素转换酶-1对RPE的影响。我们使用了大鼠RPE细胞系RPE-J。低氧应激采用氯化钴、低糖、低氧供氧。Western印迹分析和实时定量聚合酶链式反应(Real-Time-PCR)检测RPE-J中血管紧张素转换酶-1和血管内皮生长因子的表达。氯化钴或低氧低糖可促进血管内皮生长因子的表达，而血管紧张素转换酶-1的表达则显著减少。用xCELLigence系统实时监测培养过程中RPE细胞的动态变化。外源性血管内皮细胞生长因子(0.2-2 nM)显著增加细胞指数(CI)，如细胞存活率、细胞数量以及在2%氧气和无葡萄糖条件下与载体或其他浓度的血管内皮生长因子的粘附性。相反，在指定条件下，外源性血管紧张素转换酶-1(2 NM)显示较低的CI。Vasohibin-1还可降低血管内皮生长因子诱导的脑梗塞指数。这些结果在标准培养条件下是观察不到的。当我们进行MTS检测或细胞计数时，外源性Vasohibin-1也显示出类似的结果。当我们在RPE-J中转导全长Vasohibin-1cDNA时，其内部Vasohibin-1的表达水平低于对照载体转导的RPE-J。综上所述，Vasohibin-1在RPE-J上表现出与VEGF相反的结果，特别是在低氧条件下。
Vascular endothelial growth factor (VEGF) is one of the main factors for inducing choroidal neovascularization in patients with age-related macular degeneration (AMD). Retinal pigment epithelium (RPE) is one of the main sources for expressing VEGF. Vasohibin-1 is a VEGF-inducible gene in human cultured endothelial cells with antiangiogenic properties. We examined the effects of vasohibin-1 against RPE. We used rat RPE cell line, RPE-J. Cobalt chloride and low glucose and oxygen supply were used for hypoxic stress. Western blot analysis and real-time PCR were performed to detect the expression of vasohibin-1 and VEGF in the RPE-J. Cobalt chloride or low oxygen and low glucose enhanced VEGF expression, whereas statistically significant less vasohibin-1 expression was observed. RPE cell dynamics was monitored using xCELLigence System for real-time cell analysis during culture. External VEGF (0.2–2 nM) enhanced Cell Index (CI), such as cell viability, number, and adhesion to the plates significantly at 2% oxygen and no glucose when compared to those of vehicle or other concentration of VEGF. Conversely, external vasohibin-1 (2 nM) showed lower CI at the indicated condition. Vasohibin-1 also reduced VEGF-induced CI index. These results were not observed under standard culture condition. When we performed MTS assay or cell count, external vasohibin-1 also showed comparable results. When we transduced full-length vasohibin-1 cDNA in RPE-J, the internal vasohibin-1 showed less VEGF expression than that of control vector-transduced RPE-J. In summary, vasohibin-1 showed opposite results as that of VEGF on the RPE-J, especially under hypoxic condition.