MEF2C在湿性年龄相关性黄斑变性中的表达及其对脉络膜新生血管和巨噬细胞极化的影响  

Expression of MEF2C in Wet Age-related Macular Degeneration and its Effect on Choroidal Neovascularization and Macrophage Polarization

作　　者：刘旭[1] 万鹏飞[1] 杨维佳 贾俊[1] 何媛[1] 赵靖康 LIU Xu;WAN Peng-fei;YANG Wei-jia;JIA Jun;HE Yuan;ZHAO Jing-kang(Department of Ophthalmology,The Second Affiliated Hospital of Xi'an Medical University,Xi'an,Shaanxi,710038,China)

机构地区：[1]西安医学院第二附属医院眼科,陕西西安710038

出　　处：《现代生物医学进展》2023年第3期433-439,共7页Progress in Modern Biomedicine

基　　金：陕西省教育厅专项科学研究计划项目(No.19JK0758);西安医学院第二附属医院一般项目(23KY0108)。

摘　　要：目的:揭示肌细胞增强因子2C(MEF2C)在湿性年龄相关性黄斑变性(AMD)中的表达及其对脉络膜新生血管(CNV)和巨噬细胞极化的影响。方法:通过qRT-PCR法检测30例湿性AMD患者(AMD组)和30例健康体检者(健康对照组)的血清MEF2C水平。将MEF2C过表达慢病毒(MEF2C-LV组)和阴性对照过表达慢病毒(NC-LV组)转染至恒河猴脉络膜血管内皮细胞系(RF/6A)。转染后,将RF/6A细胞分为常氧组(Normoxia组)、低氧组(Hypoxia组)、低氧+NC-LV组(Hypoxia+NC-LV组)、低氧+MEF2C-LV组(Hypoxia+MEF2C-LV组)。转染及缺氧处理后,分别测定各组细胞进行Matrigel小管。通过激光诱导CNV C57BL/6J小鼠模型,将建模成功的C57BL/6J小鼠随机分为模型组、NC-LV组和MEF2C-LV组,每组10只,未建模的小鼠作为对照组。然后对NC-LV组和MEF2C-LV组小鼠玻璃体腔注射NC-LV或MEF2C-LV,对照组和模型组小鼠不进行治疗。治疗7 d后进行眼底荧光血管造影(FFA)和眼球苏木精伊红(HE)染色。通过qRT-PCR和Western blot检测MEF2C、VEGFA、VEGFR2、IL-12p35、IL-12p40和IL-10的m RNA和蛋白表达。结果:与Healthy组相比,AMD组患者的血清MEF2C水平显著降低(1.00±0.23 vs 0.48±0.29,t=7.689,P<0.001)。与Normoxia组相比,Hypoxia组的闭合管腔数量增加(P<0.05)。与Hypoxia组相比,Hypoxia+MEF2C-LV组的闭合管腔数量减少(P<0.05)。与模型组相比,MEF2C-LV组视网膜和脉络膜病变程度减轻,结构基本恢复正常,脉络膜组织厚度降低,血管生成减少。与模型组相比,MEF2C-LV组的CNV相对荧光强度降低,脉络膜组织中MEF2C、VEGFA和VEGFR2的m RNA和蛋白表达水平均降低(P<0.05)。与模型组相比,MEF2C-LV组脉络膜组织中IL-12p35和IL-12p40的m RNA和蛋白表达水平均升高,IL-10均降低(P<0.05)。结论:MEF2C在湿性AMD患者血清中低表达,上调MEF2C可抑制脉络膜血管生成,并促进巨噬细胞从M2型向M1型的转换。Objective: To reveal the expression of myocyte enhancer factor 2C(MEF2C) in wet age-related macular degeneration(AMD) and its effect on choroidal neovascularization(CNV) and macrophage polarization. Methods: The serum MEF2C levels of 30 wet AMD patients(AMD group) and 30 healthy subjects(Healthy group) were detected by qRT-PCR. The MEF2C overexpressing lentivirus(MEF2C-LV group) and negative control overexpressing lentivirus(NC-LV group) were transfected into rhesus monkey choroidal endothelial cell line(RF/6A). After transfection, RF/6A cells were divided into normoxia group(Normoxia), hypoxia group(Hypoxia),hypoxia+NC-LV group(Hypoxia+NC-LV), and hypoxia+MEF2C-LV group(Hypoxia+ MEF2C-LV). After transfection and hypoxia treatment, Matrigel tubule formation assay was performed on cells in each group. By laser-induced CNV C57BL/6J mouse model, the successfully modeled C57BL/6J mice were randomly divided into model group(Model), NC-LV group and MEF2C-LV group, 10 mice in each group. Unmodeled mice served as the Control group. Then, NC-LV or MEF2C-LV were injected into the vitreous cavity of mice in NC-LV group and MEF2C-LV group, and mice in Control group and Model group were not treated. Fundus fluorescein angiography(FFA) and ocular hematoxylin and eosin(HE) staining were performed 7 days after treatment. The m RNA and protein expressions of MEF2C, VEGFA, VEGFR2, IL-12p35, IL-12p40 and IL-10 were detected by qRT-PCR and Western blot. Results: Compared with Healthy group, the serum MEF2C levels in AMD group were significantly lower(1.00±0.23 vs 0.48±0.29, t=7.689, P<0.001). Compared with Normoxia group, the number of closed tubes was increased in Hypoxia group(P<0.05). Compared with Hypoxia group, the number of closed tubes in Hypoxia+MEF2C-LV group was decreased(P<0.05). Compared with Model group, the degree of retinal and choroidal lesions in MEF2C-LV group was alleviated, the structure basically returned to normal, the thickness of the choroidal tissue was reduced, and angiogenesis was reduced. Compared with

关 键 词：年龄相关性黄斑变性 脉络膜新生血管 肌细胞增强因子2C 血管生成 巨噬细胞极化 

分 类 号：R-33[医药卫生] R774.5