基于鹌鹑胚绒毛尿囊膜的血管生成双向干预评价模型的建立  

Establishment of evaluation model for bidirectional intervention of angiogenesis with quail chorioallantoic membrane

作　　者：崔鹤蓉 向虹俊 房康 王奥奥 王鹏龙[1] 雷海民[1] Cui Herong;Xiang Hongjun;Fang Kang;Wang Aoao;Wang Penglong;Lei Haimin(School of Chinese Materia Medica,Beijing University of Chinese Medicine,Beijing 102488,China)

机构地区：[1]北京中医药大学中药学院

出　　处：《北京中医药大学学报》2019年第10期813-818,共6页Journal of Beijing University of Traditional Chinese Medicine

基　　金：国家自然科学基金资助项目(No.81603256);中华中医药学会青年人才托举工程项目(No.CACM-2018-QNRC2-B08);中央高校基金科研业务项目(No.2019-JYB-TD005);北京中医药大学研究生自主课题(No.2019-JYB-XS-076)~~

摘　　要：目的以鹌鹑胚绒毛尿囊膜(qCAM)为研究载体,构建一种快速、直观、可量化的血管生成双向干预化合物的筛选及评价模型。方法方法学考察部分通过比较明胶海绵、分子筛、米粒、大孔树脂、硅胶粒、甲基纤维素的结果直观性、稳定性,确定常规给药载体;通过观察开窗实验后qCAM上血管的发育情况,确定给药后孵育时间;通过比较不同批次qCAM上微血管(三级+四级)的发育情况,考察模型的持续稳定性。模型的可靠性实验选择课题组前期活性可靠的化合物薯蓣皂苷元-氨基酸拼合物(DG-15)、白桦脂酸-川芎嗪-氨基酸拼合物(BA-12)作为模式药,分析不同剂量试药对qCAM血管宏观形态、血管计数的影响,评价模型的可靠性。结果方法学考察结果表明,明胶海绵既可以对给药准确定量、准确找出给药点,又可以排除载体自身对受试药物及qCAM的影响,作为本模型的给药载体;开窗实验后第2天,qCAM开始逐渐向胚胎下沉给胚胎供给营养,影响取材完整性,因此选择开窗实验后36 h作为本模型的给药时间;不同月份鹌鹑胚绒毛尿囊膜上微血管(三级+四级)计数比较结果显示,qCAM上血管发育情况基本保持稳定。模型的可靠性实验结果显示,本模型能初步评价DG-15(20μg)的促进血管生成活性(P<0.05),以及BA-12(20μg)的抑制血管生成活性(P<0.05)。结论本文建立的方法能够相对可靠地评价药物对血管生成的双向干预作用,为血管生成活性化合物的研发创制提供筛选方法与参考依据。Objective To construct a rapid,visual and quantifiable method for the screening and evaluation of compounds for bidirectional intervention of angiogenesis with quail chorioallantoic membrane(qCAM)as the carrier.Methods By comparing the results of gelatin sponge,molecular sieve,rice grain,macroporous resin,silica gel particles and methyl cellulose,the conventional drug carriers were determined;by observing the development of blood vessels on qCAM afterwindow opening test,the incubation time after administration was set;by comparing the development of microvasculature in different batches,the continuous stability of the model was tested.In the reliability test of the model,DG-15 and BA-12 were selected as model drugs,and the effects of different doses of these drugs on the macromorphology and vascular count of qCAM were analyzed evaluate to eudu ate the reliabity of tho model.Results The gelatin sponge was chosen as the drug delivery carrier as it could accurately locate the drug delivery point with exact quantity and little influence on the qCAM or the drug.36 hr after the window opening test was chosen as the best time for administration as the integrity of samples would be affected when qCAM began to gradually sink into the embryo to provide nutrition on the second day after the window opening test.By comparing the microvessel counts(grade 3+grade 4)in qCAM in different months,the development of blood vessels in qCAM was basically stable.In the reliability test,the model could be adopted to preliminarily evaluate the activity of DG-15(20 μg)in promoting angiogenesis(P<0.05),and the activity of BA-12(20μg)in inhibiting angiogenesis(P<0.05).Conclusion The method established in this study could possibly be used to evaluate the bidirectional intervention effect of drugs on angiogenesis with relative reliability,and provide a screening method and basis for the research and development of active compounds on angiogenesis.

关 键 词：血管生成 评价方法 筛选模型 鹌鹑胚绒毛尿囊膜模型 

分 类 号：R285.5[医药卫生—中药学]