中医气虚血瘀证斑马鱼模型的构建与评价  

Construction and Evaluation of Zebrafish Model with Qi Deficiency and Blood Stasis Syndrome

作　　者：郭帅杰 李思耐[1,2] 刘卫红 刘红旭[1] 周明学 GUO Shuaijie;LI Sinai;LIU Weihong;LIU Hongxu;ZHOU Mingxue(Beijing Hospital of Traditional Chinese Medicine Affiliated to Capital Medical University,Beijing 100010,China;Beijing Institute of Traditional Chinese Medicine,Beijing 100010,China)

机构地区：[1]首都医科大学附属北京中医医院,北京100010 [2]北京市中医药研究所,北京100010

出　　处：《世界中医药》2023年第6期778-782,共5页World Chinese Medicine

基　　金：国家自然科学基金项目(81673744)。

摘　　要：目的:对中医气虚血瘀证斑马鱼模型进行构建与评价。方法:随机选取90尾受精后4 d(4 dpf)野生型AB品系斑马鱼于6孔板中,每孔均处理30尾斑马鱼。分为正常对照组、气虚血瘀证组和药物验证组。采用无水亚硫酸钠和异丙肾上腺素盐酸盐联合刺激建立气虚血瘀证斑马鱼模型。药物验证组斑马鱼在造模基础上给予水溶脑心通胶囊。每组随机选取10尾斑马鱼,使用行为学分析仪分析斑马鱼的总运动距离;采用乳酸和ATP试剂盒测定斑马鱼体内乳酸和ATP水平;置于荧光显微镜下拍照,图像处理软件分析斑马鱼心脏凋亡细胞心肌细胞凋亡荧光强度;采用心跳血流分析系统分析斑马鱼血流速度和心输出量。结果:与正常对照组比较,气虚血瘀证组斑马鱼总运动距离和ATP水平均明显降低(P<0.05,P<0.001),气虚血瘀证组斑马鱼体内乳酸水平和心肌细胞凋亡荧光强度明显高于正常对照组(P<0.001,P<0.01),而血流速度和心输出量明显低于正常对照组(P<0.01,P<0.05)。与气虚血瘀证组比较,药物验证组斑马鱼总运动距离、体内ATP水平、血流速度和心输出量明显升高(P<0.05,P<0.001),斑马鱼心肌细胞凋亡荧光强度和体内乳酸水平明显降低(P<0.001)。结论:无水亚硫酸钠和异丙肾上腺素盐酸盐联合干预可成功建立气虚血瘀证斑马鱼模型。Objective:To construct and evaluate the zebrafish model with qi deficiency and blood stasis syndrome.Methods:Ninety wild-type AB strains of zebrafish with 4 days of post-fertilization(4 dpf)were randomly selected and placed in 6-well plates,and 30 zebrafish were treated in each well(experimental group).They were divided into a normal group,a syndrome group,and a verification group.The zebrafish model of qi deficiency and blood stasis syndrome was established by the combined intervention of anhydrous sodium sulfite and isoproterenol hydrochloride.Zebrafish in the verification group were given water-soluble Naoxintong Capsules based on the modeling methods.Ten zebrafish were randomly selected from each group.The total movement distance of zebrafish was analyzed by a behavioral analyzer,and the content of lactic acid and adenosine triphosphate(ATP)in zebrafish was determined by lactic acid and ATP kit.The fluorescence intensity of apoptotic cells in zebrafish hearts was analyzed by image processing software,and the blood flow velocity and cardiac output of zebrafish were analyzed by a heartbeat blood flow analysis system.Results:As compared with the normal group,the total movement distance and the content of ATP of zebrafish in the syndrome group were significantly lower(P<0.05,P<0.001).The lactic acid content and the fluorescence intensity of apoptotic cells in zebrafish hearts in the syndrome group were significantly increased than those in the normal group(P<0.001,P<0.01),and the blood flow velocity and cardiac output were significantly decreased than those in the normal group(P<0.01,P<0.05).As compared with the syndrome group,the total movement distance,the content of ATP,blood flow velocity,and cardiac output of zebrafish in the verification group were significantly increased than those in the syndrome group(P<0.05,P<0.001),while the fluorescence intensity of apoptotic cells and the content of lactic acid in vivo in zebrafish hearts were significantly decreased than those in the syndrome group(P<0.001).Conclusi

关 键 词：气虚血瘀 斑马鱼 动物模型制备 证候模型 脑心通胶囊 无水亚硫酸钠 异丙肾上腺素盐酸盐 

分 类 号：R285.5[医药卫生—中药学] R-332[医药卫生—中医学]