尼古丁通过调节巨噬细胞极化加重实验性脉络膜新生血管的形成  

Nicotine aggravates experimental choroidal neovascularization by modulating macrophage polarization

作　　者：巩亚军 赖坤贝 李龙辉 黄创新 许发宝 周立军 金陈进 GONG Ya-jun;LAI Kun-bei;LI Long-hui;HUANG Chuang-xin;XU Fa-bao;ZHOU Li-jun;JIN Chen-jin(State Key Laboratory of Ophthalmology,Zhongshan Ophthalmic Center,Sun Yat-sen University,Guangzhou 510060,China)

机构地区：[1]中山大学中山眼科中心眼科学国家重点实验室

出　　处：《中国病理生理杂志》2020年第1期104-111,共8页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.81670866; No.81600741);广东省科技计划项目(No.2017A020215038)

摘　　要：目的:探究巨噬细胞在尼古丁加重脉络膜新生血管(CNV)形成中的作用。方法:饮用尼古丁水溶液(100 mg/L)4周后,使用激光诱导小鼠CNV模型,7 d后用异硫氰酸荧光素标记的葡聚糖心脏灌注,测量CNV面积。分别于激光后1、3、7和14 d采用免疫荧光法检测巨噬细胞的时间和空间分布。采用RT-qPCR检测巨噬细胞相关分子标志物的mRNA表达。同时采用ELISA法检测激光后第1和3天视网膜色素上皮-脉络膜-巩膜组织中血管内皮生长因子(VEGF)、细胞间黏附分子1(ICAM-1)、肿瘤坏死因子α(TNF-α)和白细胞介素6(IL-6)的表达。结果:尼古丁显著增加CNV的渗漏程度及CNV的面积,尼古丁组的重度渗漏率和CNV面积分别为56.25%和(17569.96±1444.00)μm^2,显著高于对照组的31.25%和(10158.63±711.00)μm^2(P<0.05)。尼古丁促进了M2型巨噬细胞的浸润,增加了M2/M1型巨噬细胞的比率(P<0.05)。尼古丁也促进了M2型巨噬细胞相关分子标志物的mRNA表达,并且促进了VEGF、ICAM-1、TNF-α和IL-6的分泌(P<0.05)。结论:尼古丁可以促进M2型巨噬细胞的极化,增加M2/M1巨噬细胞的比例,从而促进CNV的发展。AIM:To investigate the role of macrophages in the process of nicotine aggravating choroidal neovascularization(CNV).METHODS:After administration of nicotine(100 mg/L in drinking water)for 4 weeks,the mice were used to induce CNV by laser photocoagulation.Seven days after laser injury,the mice were perfused with fluorescein isothiocyanate-labeled dextran through the left ventricle,and areas of CNV were measured.At the 1 st,3 rd,7 th and 14 th days after laser injury,the spatial and temporal distribution of macrophages was detected by immunofluorescence staining.The mRNA expression of macrophage-related markers were detected by RT-qPCR.Moreover,vascular endothelial growth factor(VEGF)and inflammation-related molecules including intercellular adhesion molecule(ICAM)-1,tumor necrosis factor(TNF)-αand interleukin(IL)-6 in the retinal pigment epithelium-choroid-sclera complexes were detected at the 1 st and 3 rd days after laser photocoagulation.RESULTS:The degree of leakage and areas of CNV were significantly increased by nicotine exposure compared with control group.Heavy leakage rate and areas of CNV of in nicotine group were 56.25%and(17569.96±1444.00)μm^2,while those in control group were 31.25%and(10158.63±711.00)μm^2(P<0.05).Nicotine promoted the infiltration of M2 macrophages and increased the ratio of M2/M1 macrophages(P<0.05).In addition,nicotine accelerated the mRNA expression of M2 macrophage-related markers and up-regulated protein expression of VEGF,ICAM-1,TNF-αand IL-6(P<0.05).CONCLUSION:Nicotine promotes the polarization of M2 macrophages,increases the ratio of M2/M1 macrophages,and ultimately accelerates the development of CNV.

关 键 词：尼古丁 巨噬细胞极化 脉络膜新生血管 炎症 

分 类 号：R774.5[医药卫生—眼科] R363.2[医药卫生—临床医学]