GPER抑制星形胶质细胞活性减少OIR小鼠视网膜新生血管生成的机制研究  

GPER-mediated inhibition of astrocyte activation mitigates retinal neovascularization in oxygen-induced retinopathy mice

作　　者：王璇 叶剑 刘玮 WANG Xuan;YE Jian;LIU Wei(Department of Ophthalmology,Army Medical Center of PLA,Chongqing,400042,China)

机构地区：[1]陆军特色医学中心眼科,重庆400042

出　　处：《陆军军医大学学报》2024年第12期1369-1377,共9页Journal of Army Medical University

基　　金：国家自然科学基金(82171054);重庆市自然科学基金(cstc2021jcyj-msxmX0164)。

摘　　要：目的探究在氧诱导下的新生小鼠视网膜缺血模型(oxygen-induced retinopathy,OIR)中,G蛋白偶联雌激素受体(G proteincoupled estrogen receptor,GPER)减少OIR小鼠视网膜新生血管生成的机制。方法42只新生小鼠采用随机数字表法分为4组:常氧对照组(n=11)、OIR组(n=11)、G-1(GPER激动剂)组(n=10)和溶剂对照组(n=10)。出生后17 d(P17)时,眼球冰冻切片免疫荧光染色观察常氧对照组小鼠GPER在视网膜分布情况。G-1组和溶剂对照组在P12~P15时分别腹腔注射给予G-1[50μg/(kg·d)]或玉米油溶剂,P17时视网膜铺片免疫荧光染色观察视网膜血管标志物(IB4)、星形胶质细胞标志物胶质纤维酸性蛋白(glia fibrilary acidic protein,GFAP)表达,蛋白免疫印迹法定量检测视网膜血管内皮生长因子A(vascular endothelial growth factor A,VEGFA)、GFAP、炎症因子TNF-α、IGF-1、IL1-β蛋白表达情况。结果GPER存在于小鼠视网膜全层,在视网膜神经节细胞层与星形胶质细胞标志物GFAP共染。与常氧对照组相比,OIR组小鼠视网膜出现新生血管与无血管区,且GPER、VEGFA和GFAP表达增多,差异具有统计学意义(P<0.05);与溶剂对照组相比,G-1组视网膜新生血管生成减少,VEGFA、GFAP、TNF-α、IGF-1、IL1-β蛋白表达下降,差异具有统计学意义(P<0.05)。结论GPER能抑制星形胶质细胞活性,减少VEGFA、炎症因子释放,减少OIR小鼠视网膜新生血管生成。Objective To investigate the mechanism by which G protein-coupled estrogen receptor(GPER)reduces retinal neovascularisation in oxygen-induced retinopathy(OIR)in newborn mice.Methods A total of 42 newborn mice were randomly divided into normoxic control group(n=11),OIR group(n=11),G-1(GPER agonist)group(n=10),and solvent control group(n=10).On postnatal day 17(P17),the distribution of GPER in the retina of mice in the normoxic control group was observed by immunofluorescence staining on frozen sections of the eyeballs.The mice of the G-1 group and solvent control group were given by intraperitoneal injection 50μg/(kg·d)G-1 or corn oil solvent from P12 to P15.Immunofluorescence staining of retinal spreads at P17 was performed to observe the expression of retinal vascular marker IB4 and astrocyte marker glia fibrilary acidic protein(GFAP).Western blotting was used to quantify the expression of retinal vascular endothelial growth factor A(VEGFA),GFAP,and inflammatory factors TNF-α,IGF-1,and IL1-β.Results GPER was present throughout the retina and co-stained with the astrocyte marker GFAP in the ganglion cell layer(GCL).Compared with the normoxic control group,the retinas of mice in the OIR group showed neovascular and avascular areas,and the expression of VEGFA and GFAP was significantly increased(P<0.05).Compared with the solvent control group,the retinal neovascularisation was reduced in the G-1 group,and the expression levels of VEGFA,GFAP,TNF-α,IGF-1,and IL1-βproteins were decreased significantly(P<0.05).Conclusion GPER inhibits astrocyte activity,reduces VEGFA expression and release of inflammatory factors,and then decreases retinal neovascularisation in OIR mice.

关 键 词：早产儿视网膜病变 氧诱导下的新生小鼠视网膜缺血模型 星形胶质细胞 炎症因子 G蛋白偶联雌激素受体 

分 类 号：R364.33[医药卫生—病理学] R774.1[医药卫生—基础医学] R977.6