中华眼底病杂志

中华眼底病杂志

Notch受体蛋白1、Delta样配体4在增生型糖尿病视网膜病变新生血管形成中的作用

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目的 观察Notch受体蛋白1(Notch1)、Delta样配体4(Dll4)在增生型糖尿病视网膜病变(PDR)纤维血管膜中的表达,初步探讨Notch1、Dll4在PDR新生血管形成中的作用及与血管内皮生长因子(VEGF)信号通路的关系。 方法 行玻璃体切割手术的PDR患者57例60只眼纳入研究。根据手术前是否行抗VEGF药物玻璃体腔注射将患眼分为PDR未注药组和PDR注药组,分别为30例32只眼、27例28只眼。PDR注药组患眼手术前2~7 d行雷珠单抗玻璃体腔注射治疗。选取同期非糖尿病伴特发性黄斑前膜患者18例18只眼作为对照组。于玻璃体切割手术中获得PDR纤维血管膜病理标本及黄斑前膜病理标本。应用荧光定量聚合酶链反应(RT-PCR)及免疫组织化学染色法检测各组病理标本中Notch1、Dll4、VEGF受体2(VEGFR2)的表达;采用苏木精-伊红染色计数突破内界膜的血管内皮细胞数量。 结果 免疫组织化学染色结果显示,PDR注药组、PDR未注药组病理标本中均有Notch1、Dll4、VEGFR2蛋白表达。PDR未注药组病理标本中Notch1、Dll4、VEGFR2蛋白相对表达量均高于PDR注药组,差异均有统计学意义(t=3.45、6.01、4.08,P=0.030、0.008、0.023)。PDR未注药组、PDR注药组病理标本中,突破内界膜的血管内皮细胞数量分别为(41.50±5.57)、(17.17±2.48)个,差异有统计学意义(t=9.58,P<0.05)。RT-PCR检测结果显示,PDR未注药组(H=12.50)、PDR注药组(H=12.50)、对照组(H=12.02)病理标本中Notch1、Dll4、VEGFR2 mRNA相对表达量比较,差异均有统计学意义(P<0.005)。其中,PDR注药组、PDR未注药组病理标本中Notch1、Dll4、VEGFR2 mRNA相对表达量显著高于对照组;与PDR未注药组比较,PDR注药组病理标本中Notch1、Dll4、VEGFR2 mRNA相对表达量均减少。Spearman相关性分析结果显示,VEGFR2与Dll4(r=0.83)、VEGFR2与Notch1(r=0.81)、Notch1与Dll4(r=0.87)之间mRNA相对表达量均呈显著正相关(P<0.05)。 结论 Notch1、Dll4在PDR纤维血管膜中的表达均高于对照组,且与VEGFR2的表达呈正相关;Notch1、Dll4在PDR新生血管的发生中发挥调节作用,其作用途径可能与VEGFR2相关。

Objective o observe the expression of Notch1 and Delta-like ligand 4 (Dll4) on the fibrovascular membranes in proliferative diabetic retinopathy (PDR), and investigate its relationship with vascular endothelial growth factor receptor 2 (VEGFR2). Methods Fifty-seven PDR patients (60 eyes) who underwent vitrectomy were enrolled in this study. The PDR patients were divided into non-injection group (30 patients, 32 eyes) and injection group (27 patients, 28 eyes). The eyes in injection group received intravitreal injection with ranibizumab at 2 to 7 days before surgery. The preretinal fibrovascular membranes were obtained from the PDR patients during vitrectomy. Eighteen epiretinal membranes were obtained from the non-diabetic patients was served as controls. The real-time polymerase chain reaction (RT-PCR) and immunohistochemical methods were used to detecting the expression of Notch1, Dll4 and VEGFR2. In the meantime, the numbers of the nucleus of vascular endothelial cells in the membranes stained with hematoxylin were counted. Results The immunohistochemical staining revealed that there were positive expression of Notch1, Dll4 and VEGFR2 in all PDR membranes, regardless of the injection of the ranibizumab. The levels of Notch1, Dll4 and VEGFR2 protein in non-injection group were higher than those of injection group (t=3.45, 6.01, 4.08;P=0.030, 0.008, 0.023). In injection group, the number of endothelial cells in the membranes reduced (17.17±2.48) compared with that of the non-injection group (41.50±5.57). There was significant difference in the number of endothelial cells in the membranes between the two groups (t=9.58,P<0.05). RT-PCR showed that the differences of the mRNA expression of Notch1, Dll4 and VEGFR2 were all statistically significant among the PDR group and control group (H=12.50, 12.50, 12.02;P<0.05).The expression of Notch1, Dll4 and VEGFR2 in the PDR membranes was higher than that of epiretinal membranes from non-diabetic patients. In the PDR group, the expression of Notch1, Dll4 and VEGFR2 of non-injection group was higher than that of injection group. Spearman correlation analysis showed that the expression of mRNA between VEGFR2 and Dll4 (r=0.83), VEGFR2 and Notch1 (r=0.81), Notch1 and Dll4 (r=0.87) were all significantly correlated (P<0.05). Conclusions The expression of Notch1 and Dll4 in the PDR membranes are higher than that of the control group, and it is positively correlated with the expression of the VEGFR2. Notch1 and Dll4 play a regulatory rule in the neovascularization in PDR, the acting way may be correlated with VEGFR2.

关键词: 视网膜新生血管化/病因学; 糖尿病视网膜病变/病理学; 受体,Notch1; 血管内皮生长因子受体2

Key words: retinal neovascularization/etiology; Diabetic retinopathy/pathology; Receptor, Notch1; Vascular endothelial growth factor receptor-2

引用本文: 郭庆敏, 孟旭霞, 刘鹏辉, 周贤慧. Notch受体蛋白1、Delta样配体4在增生型糖尿病视网膜病变新生血管形成中的作用. 中华眼底病杂志, 2017, 33(3): 275-280. doi: 10.3760/cma.j.issn.1005-1015.2017.03.013 复制

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