摘要
目的观察红细胞生成素（EPO）在新生血管性青光眼（NVG）中的表达及体外血管诱导形成作用。方法采用酶联免疫吸附法（ELISA）检测2014年6至10月在内蒙古医科大学附属医院眼科就诊的10例NVG患者及10例非NVG患者房水及玻璃体中的EPO水平。将人脐静脉内皮细胞（HUVECs）作为研究对象，依据EPO浓度分为0.1、1、10 U/ml EPO组及10 ng/ml血管内皮生长因子（VEGF）组处理HUVECs细胞，观察不同浓度的重组人EPO及VEGF对体外HUVECs增殖，血管管腔样结构生成以及毛细血管样结构生成作用的影响，同时采用Western印迹及实时定量PCR（RT－PCR）检测细胞中低氧诱导因子－1α（HIF－1α）蛋白及mRNA表达水平。结果本研究中未见中止试验的患者，非NVG患者眼房水EPO水平为（29.4±0.6）ng/L，而NVG患者房水中EPO含量高达（307.5±7.6）ng/L，二者比较差异有统计学意义（t＝2.186，P＝0.039）；非NVG患者眼玻璃体中EPO水平为（16.7±1.2）ng/L，而NVG患者玻璃体中EPO含量异常增高达到（267.5±11.4）ng/L，二者比较差异有统计学意义（t＝2.517，P＝0.019）。与对照组比较，0.1、1、10 U/ml EPO及10 ng/ml VEGF能显著促进HUVECs增殖，0.1、1、10 U/ml EPO及10 ng/ml VEGF能促进HUVECs体外血管管腔样结构生成以及毛细血管样结构生成，同时降低HIF－1α蛋白及mRNA表达，差异均有统计学意义（均P〈0.05）。结论EPO在NVG中高表达，且外源性EPO能显著促进HUVECs增殖，血管管腔样结构及毛细血管样结构生成，可能与提高HIF－1α表达有关。
Objective To explore the expression of erythropoietin （EPO） in neovascular glaucoma （NVG） and its role in the angiogenesis in vitro.Methods The levels of EPO in aqueous humor and vitreous body of NVG patients （10 cases） and non-NVG patients （10 cases） were detected by enzyme-linked immunosorbent assay （ELISA）. Human umbilical vein endothelial cells （HUVECs） were used as the research object, and were divided into EPO 0.1, 1, 10 U/ml group according to EPO concentration, on the basis of treatment of HUVECs cells with 10 ng/ml vascular endothelial growth factor （VEGF）. Effect of different concentrations of recombinant human EPO and VEGF on the proliferation, angiogenesis and the formation of capillary like structures of HUVECs in vitro was explored, meanwhile, protein and mRNA expression of hypoxia inducible factor-1α （HIF-1α） was detected by Western blot and RT-PCR.Results In the study, no patients discontinued the trial. EPO level of aqueous humor in non-NVG patients was （29.4±0.6） ng/L, while EPO level in aqueous humor of NVG patients reached （307.5±7.6） ng/L, and the difference was statistically significant （t=2.186, P=0.039）; vitreous EPO levels in non-NVG patients was （16.7±1.2） ng/L, while vitreous EPO levels in NVG patients reached （267.5±11.4） ng/L, with a statistically significant difference （t=2.517, P=0.019）. Compared with control group, 0.1, 1, 10 U/ml EPO and 10 ng/ml VEGF promoted the proliferation of HUVECs, and formation of vascular lumen-like and capillary-like structure in vitro, and up-regulated the protein and mRNA expression of HIF-1α （all P〈0.05）.Conclusion EPO was highly expressed in the NVG patients, and exogenous EPO could significantly promote the proliferation of HUVEC and formation of vascular lumen-like and capillary-like structure, which may be related to up-regulation of HIF-1 expression.
出处
《中华医学杂志》
CAS
CSCD
北大核心
2016年第46期3729-3734,共6页
National Medical Journal of China
