Page 59 - 2020年2月第31卷第3期
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Study on the Heart Protective Effect of Panax quinquefolium Extract Based on Zebra Fish Model
LYU Jing,LI Chen,YANG Longfei,GAO Yan,ZHAO Bonian(College of Pharmacy,Shandong University of
TCM,Jinan 250355,China)
ABSTRACT OBJECTIVE:To study the heart protective effect of Panax quinquefolium extract of Zebra fish. METHODS:The
drug dosage of P. quinquefolium extract and the optimal modeling dose of terfenadine were determined by tolerance test;24 hours
post fertilization (24 hpf),Zebra fish were divided into control group (0.5% dimethyl sulfoxide),model group (2.5 μ g/mL
terfenadine), positive control group (2.5 μ g/mL terfenadine + 30 μ g/mL resveratrol) and different mass concentration P.
quinquefolium extract groups(2.5 μg/mL terfenadine+5,10,25 μg/mL P. quinquefolium extract,by raw material). 48 h after
incubation,the heart rate and sinus venous-bulbus arteriosus(SV-BA)interval of Zebra fish were used as indexes,and the heart
injury repair rate was calculated to investigate the heart protection of P. quinquefolium extract. Another 24 hpf Zebra fish were
divided into blank control group,model group,positive control group,20 batches of P. quinquefolium extract from different
producing area groups(5 μg/mL,by raw material),with same administration route and 48 h of incubation. The heart rate,SV-BA
interval of heart was determined,heart injury repair rate and heart injury relative repair rate were calculated;the heart protective
effect of P. quinquefolium extract was further validated. RESULTS:Compared with blank control group,heart rate was decreased
significantly (P<0.01),while SV-BA interval was increased significantly in model group (P<0.01). Compared with model
group,heart rate of Zebra fish was increased significantly in positive control group and 5,10,25 μg/mL P. quinquefolium extract
groups(P<0.01),while SV-BA interval of Zebra fish was shortened significantly in 5,10 μg/mL P. quinquefolium extract groups
(P<0.05 or P<0.01);heart injury repair rate of Zebra fish in 5 μ g/mL P. quinquefolium extract group reached 73.77% . In
validation test,compared with model group,heart rate was increased significantly(P<0.01),while SV-BA interval of Zebra fish
was decreased significantly in 20 batches of P. quinquefolium extract groups (P<0.01); heart injury repair rate ranged
70.45%-85.78%,and heart injury relative repair rate ranged 75.48%-98.12%. CONCLUSIONS:P. quinquefolium extract has good
heart protective effects.
KEYWORDS Panax quinquefolium;Zebra fish;Heart protection;Sinus venous-bulbus arteriosus;Interval;Heart rate
西洋参为五加科植物西洋参(Panax quinquefolium 电热鼓风干燥箱(上海博讯有限公司);KQ2200DV型超
L.)的干燥根,其味甘、微苦,性凉,归心、肺、肾经 ,可补 声波清洗仪(昆山市超声仪器有限公司);AXIO Zoom.
[1]
[2]
气养阴 ,常用于治疗气虚阴亏型冠心病 [3-4] 。相关研究 V16型体式显微镜(德国Carl Zeiss公司);FW-80型高速
[5]
表明,西洋参通过益气养阴、和血来补益心气 ,可改善 万能粉碎机(北京市永光明医疗仪器有限公司)。
冠心病患者经皮冠状动脉介入治疗(PCI)术后的胸闷、 1.2 药品与试剂
心悸、乏力等症状,提高患者生活质量 。但关于西洋参 20 批西洋参药材购自安徽省亳州市中药材交易市
[6]
是否均具有心脏保护作用尚不明确,而且目前常用的心 场,经山东省中医药研究院林慧彬研究员鉴定均为真
脏功能研究动物模型造模过程操作复杂,不便于进行在 品,其总皂苷含量均符合2015年版《中国药典》(一部) [1]
体的心脏结构及功能研究,也不适宜进行高通量药材心 规定;特非那定标准品、链酶蛋白酶E(北京索莱宝科技
脏保护活性筛选。斑马鱼是国际认可的新型模式生物 , 有限公司,批号:607A023、515S021,纯度:≥98%);白藜
[7]
具有体型小、心脏发育周期全透明、繁殖周期短、产卵量 芦醇标准品(成都普思生物科技股份有限公司,批号:
[8]
大、发育迅速等优点 。基于此,本研究采用特非那定复 PS000122,纯度:≥98%);其他试剂均为市售分析纯,水
制斑马鱼心脏损伤模型 [9-10] ,以心脏静脉窦-动脉球 为蒸馏水。20批西洋参样品信息见表1。
(SV-BA)间距为主要指标、心率为辅助指标探讨西洋参 1.3 动物
提取物的心脏保护作用,并进一步为西洋参提取物心脏 野生型AB品系斑马鱼,雌雄兼有,由山东省科学院
保护作用谱-效相关质量评价系统的建立提供药效学 生物所提供。在专用的养殖房内,斑马鱼采用明暗交替
基础。 方式饲养(光照14 h后避光10 h),循环水及环境温度控
1 材料 制为(28.0±0.5)℃,并按雌雄分缸饲养,每日饲喂丰
1.1 仪器 年虾。
SPX-280B-G 型光照培养箱(上海基星生物科技有 2 方法与结果
限公司);AGBP210S 型电子天平(德国 Sartorius 公司); 2.1 西洋参提取物母液的制备
Milli-Q IQ7000 型纯水机(美国 Millipore 公司);KDM-A 取西洋参粉末(过三号筛)1 g,精密称定,置于具塞
型控温电热套(金坛市医疗仪器厂);GZX-9140MBE 型 圆底烧瓶中,精密加入70%乙醇50 mL,称定质量,加热
中国药房 2020年第31卷第3期 China Pharmacy 2020 Vol. 31 No. 3 ·309 ·