血管内皮钙黏蛋白在实验性角膜新生血管形成中的抗凋亡作用
Anti-apoptosis effects of vascular endothelial cadherin in experimental corneal neovascularization.
作者信息
Liu Gao-Qin, Wu Hong-Ya, Xu Jing, Wang Meng-Jiao, Lu Pei-Rong, Zhang Xue-Guang
机构信息
Department of Ophthalmology, the First Affiliated Hospital of Soochow University, Suzhou 215006, Jiangsu Province, China ; Jiangsu Clinical Immunology Institute, the First Affiliated Hospital of Soochow University, Suzhou 215006, Jiangsu Province, China.
Jiangsu Clinical Immunology Institute, the First Affiliated Hospital of Soochow University, Suzhou 215006, Jiangsu Province, China.
出版信息
Int J Ophthalmol. 2015 Dec 18;8(6):1083-8. doi: 10.3980/j.issn.2222-3959.2015.06.01. eCollection 2015.
AIM
To explore the effects and mechanism of vascular endothelial cadherin (VE-cadherin) on experimental corneal neovascularization (CRNV).
METHODS
Mouse corneas were burned with sodium hydroxide to build a CRNV model. The burned corneas were locally administrated with anti-mouse VE-cadherin neutralizing antibody. Annexin V and cluster of differentiation 31 (CD31) double staining was used to measure vascular endothelial cell apoptosis with the use of flow cytometry (FCM). The protein expression of NADPH oxidase 2 (Nox2), caspase-3, and protein kinase C (PKC) in the burned corneas were examined by Western blot. Human retinal endothelial cell (HREC) proliferation was detected using a Cell Counting Kit 8 (CCK-8) assay in vitro.
RESULTS
The amount of CRNV peaked two weeks after the alkali burn. FCM confirmed that VE-cadherin neutralizing antibody treatment increased CD31 positive cell apoptosis. Western blot revealed that the intracorneal protein expression of Nox2 and caspase-3 were up-regulated, while PKC was down-regulated in the VE-cadherin neutralizing antibody administrated group. CCK-8 assay showed that VE-cadherin neutralizing antibody markedly inhibited HREC proliferation.
CONCLUSION
VE-cadherin exhibited an anti-apoptosis effect through enhanced PKC signaling and an enhanced cell proliferation pathway.
目的
探讨血管内皮钙黏蛋白（VE-钙黏蛋白）对实验性角膜新生血管化（CRNV）的影响及机制。
方法
用氢氧化钠烧灼小鼠角膜建立CRNV模型。对烧灼后的角膜局部给予抗小鼠VE-钙黏蛋白中和抗体。采用膜联蛋白V和分化簇31（CD31）双重染色，利用流式细胞术（FCM）检测血管内皮细胞凋亡。通过蛋白质印迹法检测烧灼后角膜中NADPH氧化酶2（Nox2）、半胱天冬酶-3和蛋白激酶C（PKC）的蛋白表达。在体外使用细胞计数试剂盒8（CCK-8）检测人视网膜内皮细胞（HREC）增殖。
结果
碱烧伤后两周CRNV量达到峰值。FCM证实，VE-钙黏蛋白中和抗体治疗增加了CD31阳性细胞凋亡。蛋白质印迹法显示，在给予VE-钙黏蛋白中和抗体的组中，角膜内Nox2和半胱天冬酶-3的蛋白表达上调，而PKC下调。CCK-8检测显示，VE-钙黏蛋白中和抗体显著抑制HREC增殖。
结论
VE-钙黏蛋白通过增强PKC信号传导和增强细胞增殖途径发挥抗凋亡作用。
Zotero 插件