三七总皂苷对新生血管性视网膜病变的干预效应及机制研究  

Intervention Effect and Mechanism of Panax Notoginseng Saponins on Neovascular Retinopathy

作　　者：金茹娜[1] 郭红 徐静[1,2] 杜霄烨[1,2] 张腾 陈瑜[1,2] Jin Runa;Guo Hong;Xu Jing;Du Xiaoye;Zhang Teng;Chen Yu(Yueyang Hospital of Integrative Traditional Chinese and Western Medicine,Shanghai 200437,China;Clinical Research Institute of Integrative Medicine,Shanghai Academy of Traditional Chinese Medicine,Shanghai 200437,China;Yueyang Clinical Medical College,Shanghai University of Traditional Chinese Medicine,Shanghai 201210,China)

机构地区：[1]上海中医药大学附属岳阳中西医结合医院,上海200437 [2]上海市中医药研究院中西医结合临床研究所,上海200437 [3]上海中医药大学岳阳临床医学院,上海201210

出　　处：《世界科学技术-中医药现代化》2021年第1期108-116,共9页Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology

基　　金：上海市科学技术委员会上海市优秀学术带头人计划(19XD1403700):三七皂苷干预视网膜新生血管的优势效应机制研究,负责人:陈瑜;上海市教育委员会上海市高校特聘教授(东方学者)跟踪计划(沪教委[人]201764):视网膜退行性病变的中医药干预及效应机制研究,负责人:陈瑜;上海中医药大学附属岳阳中西医结合医院科研项目(2017YJ08):金钗石斛多糖对STZ小鼠视网膜早期炎症反应的作用机制研究,负责人:金茹娜。

摘　　要：目的探讨三七总皂苷(Panax notoginseng saponins,PNS)对视网膜新生血管性病变进程中视网膜神经血管单元病理改变的干预效应和机制,为三七及三七皂苷制剂临床应用于相关视网膜病变的治疗提供部分实验和理论依据。方法本课题采用氧诱导视网膜病变(Oxygen induced retinopathy,OIR)小鼠模型,将7天龄(Postnatal 7,P7)C57/BL6J小鼠根据体质量随机分为正常对照组(NC组),OIR模型组(OIR组)和PNS治疗组(PNS组),造模后从P12开始腹腔注射给药,根据不同的实验内容调整给药天数。于P7、P12、P19、P26和P33测量小鼠的体质量,于P19和P33应用小动物视网膜成像显微镜活体采集视网膜及血管图像,并定量分析;于P33应用全视网膜眼电图Ganzfeld ERG系统进行小鼠视网膜电图波形采集,对a波和b波振幅进行分析;于P46和P66将小鼠处死,取眼球制作眼杯,冰冻切片后进行胶质纤维酸性蛋白(Glial fibrillary acidic protein,GFAP)免疫组织化学分析;于P13和P15处死小鼠,取视网膜组织,采用Real-time PCR分析各组视网膜抗血管内皮生长因子A(Vascular endothelial growth factor A,VEGFA)、血管内皮细胞钙粘蛋白(Vascular endothelial-cadherin,VE-cadherin)、肿瘤坏死因子-α(Tumor necrosis factor,TNF-α)、白介素-1β(interleukin-1β,IL^(-1)β)的mRNA表达水平。结果(1)NC组、OIR组和PNS组的小鼠在生长发育中不同天龄的体质量无统计学差异(P>0.05)。(2)P19 OIR组小鼠视网膜血管荧光素钠渗漏面积显著大于NC组(P<0.05),而PNS组渗漏面积显著小于OIR组(P<0.05)。(3)P33 OIR组小鼠视网膜动脉迂曲指数(Retinal artery tortuosity index,RAT index)显著大于NC组(P<0.05),而PNS组显著小于OIR组(P<0.05)。(4)OIR组小鼠b波振幅与NC组相比有统计学差异(P<0.05);PNS组b波振幅与OIR组相比有统计学差异(P<0.05)。(5)P46和P66 OIR组小鼠视网膜Müller细胞GFAP免疫阳性明显增强,并可达内核层和外核层,PNS组GFAP免疫阳性和NC组相近,Objective To investigate the intervention effect and mechanism of Panax notoginseng saponins(PNS) on the pathological changes of the retinal neurovascular unit in the process of retinal neovascular lesions, and to provide some experimental and theoretical basis for the treatment by Radix Notoginseng and panax notoginseng saponins preparations.Methods The oxygen-induced retinopathy(OIR) mouse model was used. Seven-day-old(Postnatal 7, P7) C57/BL6 J mice were randomly divided into normal control group(NC group), OIR model group(OIR group) and PNS treatment group(PNS group) according to body weight. The intraperitoneal injection was carried out from P12 after modeling. The administration days were adjusted according to different experimental content. The weights of mice were measured at P7,P12, P19, P26 and P33, and small animal retinal imaging microscopes were used to collect retinal and blood vessel images in vivo at P19 and P33, and quantitative analysis was performed. At P33, the full-retinal electroencephalogram Ganzfeld ERG system was used for the acquisition of mice electroretinogram waveforms and analysis of a-wave and bwave amplitudes. At P46 and P66, the mice were sacrificed and the eyeballs were taken to make eyecups, the glial fibrillary acidic protein(GFAP) immunization was performed after frozen sections histochemical analysis. The mice were sacrificed at P13 and P15, retinal tissue was taken, and retinal anti-vascular endothelial growth factor A(VEGFA) and vascular endothelial-cadherin(VE-cadherin), tumor necrosis factor-α(TNF-α), interleukin-1β(IL^(-1)β) mRNA expression levels were analyzed by Real-time PCR. Results (1) There was no statistically significant difference in body weight between mice of NC group, OIR group and PNS group at different ages during growth and development(P > 0.05).(2) The retinal vascular fluorescein sodium leakage area of mice in the P19 OIR group was significantly larger than that in the NC group(P < 0.05), while the leakage area of the PNS group was significantly smal

关 键 词：三七总皂苷 新生血管性视网膜病变 神经血管单元 血管渗漏 炎症 

分 类 号：R276.7[医药卫生—中医五官科学]