血管内皮生长因子荧光共聚焦显微图像分析  

The analysis of vascular endothelial growth factor fluorescence confocal microscopy images

作　　者：黄妍 丁云鹤[1,2] 赵诣深 张璐 HUANG Yan;DING Yunhe;ZHAO Yishen;ZHANG Lu(School of Biomedical Engineering,Capital Medical University,Beijing 100069;Beijing Key Laboratory of Fundamental Research on Biomechanics in Clinical Application,Beijing 100069)

机构地区：[1]首都医科大学生物医学工程学院,北京100069 [2]首都医科大学临床生物力学应用基础研究北京市重点实验室,北京100069

出　　处：《北京生物医学工程》2025年第1期1-8,共8页Beijing Biomedical Engineering

基　　金：北京市教育委员会科学研究计划项目(KM202010025026)资助。

摘　　要：目的建立一种血管内皮生长因子(vascular endothelial growth factor,VEGF)在视网膜内异常表达情况的可视化和定量评估方法,为糖尿病视网膜病变(diabetic retinopathy,DR)的早期检测和治疗提供帮助。方法对大鼠视网膜铺片进行免疫荧光染色和成像。对VEGF和微血管病变区域进行分割、提取与三维重建,采用VEGF阳性单位、微血管密度、分形维数等定量参数对结果进行评估,比较糖尿病视网膜病变8周模型组(n=10)与正常对照组(n=10)的差异。结果在DR病变模型中,VEGF探针高表达区域与血管异常区域存在共定位,VEGF聚集于视网膜微血管分叉或膨大的血管壁表面。DR组的微血管分形维数、密度和三维血管弯曲度明显高于正常对照组(P<0.05);VEGF阳性单位(positive unit,PU)值在DR组中明显高于对比组(P<0.05);微血管空隙度差异无统计学意义(P>0.05)。结论本研究提出的VEGF定量评估方法,能够定性和定量地反映VEGF在DR模型视网膜中的异常分布,为抗VEGF药物研究提供定量及可视化手段。Object To establish a method for visualizing and quantifying the abnormal expression of vascular endothelial growth factor(VEGF)in the retina,and to help the early detection and treatment of diabetic retinopathy(DR).Methods Immunofluorescence staining and imaging were performed on the rat retinal flatmounts.VEGF and microvessels were segmented,extracted and reconstructed.Quantitative parameters such as positive VEGF units,microvascular density and fractal dimension were used and the differences between the 8-week diabetic retinopathy model group(n=10)and the normal control group(n=10)were compared.Results In the DR model,there was colocalization between the high expression region of VEGF probe and the abnormal region of vascular,and VEGF accumulates on the surface of retinal microvascular bifurcation or swollen vessel wall.The fractal dimension,density and curvature of microvessels in DR group were significantly higher than those in normal control group(P<0.05).The positive unit(PU)value of VEGF in DR group was significantly higher than that in comparison group(P<0.05).There was no significant difference in microvascular void(P>0.05).Conclusions In this study,a quantitative evaluation method of VEGF is proposed,which can reflect the abnormal distribution of VEGF in the retina of DR model qualitatively and quantitatively,and provide a quantitative and visual way for the research of anti-VEGF drugs.

关 键 词：糖尿病视网膜病变 血管 荧光成像 定量分析 图像重建 

分 类 号：R318.04[医药卫生—生物医学工程]