基于巨噬细胞和胶质细胞探讨明睛颗粒对湿性年龄相关性黄斑变性纤维血管膜的影响  

Effect of Mingjing Granules on Fibrovascular Membrane of Wet Age-related Macular Degeneration Based on Macrophages and Glial Cells

作　　者：李晓宇 梁丽娜[1] 高云 LI Xiaoyu;LIANG Lina;GAO Yun(Eye Hospital,China Academy of Chinese Medical Sciences,Beijing 100040,China)

机构地区：[1]中国中医科学院眼科医院,北京100040

出　　处：《中国实验方剂学杂志》2023年第22期95-103,共9页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(81973912,82274589);中国中医科学院科技创新工程项目(CI2021A02602);中国中医科学院委托项目(ZZ13-036-1);首都卫生发展科研专项(2020-2-4183);中国中医药循证医学中心“业务研究室主任专项”(2020YJSZX-2);中医药循证能力建设项目(1010304)。

摘　　要：目的:基于巨噬细胞和胶质细胞探讨明睛颗粒(MJKL)对实验性湿性年龄相关性黄斑变性(nAMD)纤维血管膜的影响,进一步阐释MJKL治疗nAMD的作用机制。方法:通过两阶段激光光凝建立实验性nAMD纤维血管膜模型,将造模成功后的BN大鼠随机分为模型组、抗血管内皮生长因子(VEGF)组(5μL/眼)、MJKL+抗VEGF组(1 g·mL^(-1)+5μL/眼)3组。模型组,给予蒸馏水灌胃;抗VEGF组,予玻璃体腔注射雷珠单抗注射液;MJKL+抗VEGF组,予玻璃体腔注射雷珠单抗注射液,同时开始予MJKL灌胃。10只正常BN大鼠不造模,常规饲养作为正常组。造模40 d后,采用眼底照相(FP)、眼底血管荧光造影(FFA)、光学相干断层扫描(OCT)、视网膜色素上皮(RPE)-脉络膜-巩膜铺片观察眼底病变形态、病变渗出面积及吸光度A;组织病理学观察视网膜结构的改变,免疫荧光法检测F4/80、离子钙接头蛋白抗原(Iba-1)、胶质纤维酸性蛋白(GFAP)的表达及分布,实时荧光定量聚合酶链反应(Reat-time PCR)检测F4/80、Iba-1、GFAP mRNA的相对表达量。结果:两阶段激光造模后40 d建立纤维血管膜模型;与模型组比较,抗VEGF组在病变渗出面积、A、病变高度、病变面积均明显降低(P<0.05),视网膜结构损伤程度显著改善;而MJKL+抗VEGF组较抗VEGF组在渗出面积、A、病变高度、病变面积明显降低(P<0.05),病变面积及视网膜结构损伤程度改善更为明显。与正常组比较,模型组F4/80与Iba-1的荧光强度均明显升高(P<0.05);与模型组比较,抗VEGF组F4/80与Iba-1的荧光强度均明显降低(P<0.05);与抗VEGF组比较,MJKL+抗VEGF组F4/80与Iba-1的荧光强度明显降低(P<0.05)。与正常组比较,模型组GFAP的荧光强度明显升高(P<0.05);与模型组比较,抗VEGF组GFAP的荧光强度明显降低(P<0.05)。与正常组比较,模型组F4/80、Iba-1、GFAP mRNA的相对表达量均明显升高(P<0.05);与模型组比较,抗VEGF组F4/80、Iba-1、GFAP mRNA的相对表达量均�Objective:To investigate the effects of Mingjing granules(MJKL)on the fibrovascular membrane of experimental wet age-related macular degeneration(nAMD)based on macrophages and glial cells and further explain the mechanism of MJKL in the treatment of nAMD.Method:The experimental nAMD fibrovascular membrane model was established by two-stage laser photocoagulation.BN rats were randomly divided into three groups:model group,anti-vascular endothelial growth factor(VEGF)group,and MJKL+anti-VEGF group.The model group was given distilled water for intragastric administration.Anti-VEGF group was injected with leizumab injection in the vitreous cavity.MJKL+anti-VEGF group was injected with leizumab injection in the vitreous cavity,and MJKL was intragastrically administered.Ten normal BN rats were not modeled and fed as controls.After 40 days of model making,fundus lesion morphology,lesion exudation area,and MD value were observed by fundus photography(FP),fundus angiography(FFA),optical coherence tomography(OCT),and retinal pigment epithelium(RPE)-choroid-sclera film.The changes in retinal structure were observed by histopathology,and the expression and distribution of F4/80,Iba-1,and GFAP were detected by immunofluorescence.The relative expression levels of F4/80,Iba-1,and GFAP mRNA were detected by realtime fluorescence quantitative polymerase chain reaction(Real-time PCR).Result:The fibrovascular membrane model was established 40d after two-stage laser modeling.The lesion exudation area,MD value,lesion height,and lesion area in the anti-VEGF group were significantly lower than those in the model group(P<0.05),and the retinal structural damage degree was significantly improved.Compared with the anti-VEGF group,the MJKL+anti-VEGF group significantly decreased the MD value,lesion height,and lesion area(P<0.05),and lesion area and retinal structural damage degree were significantly improved.The fluorescence intensity of F4/80 and Iba-1 in the model group was significantly higher than that in the normal group(P<0.05),and th

关 键 词：明睛颗粒 两阶段激光诱导 纤维血管膜 巨噬细胞 胶质细胞 

分 类 号：R284[医药卫生—中药学] R285[医药卫生—中医学] R289R287R22R2-031R33R24