基于斑马鱼模型的朝鲜淫羊藿抗骨质疏松活性部位筛选  

Screening on antiosteoporotic active parts of Epimedium koreanum based on zebrafish model

作　　者：詹扬[1,2] 韦英杰[1,2] 孙娥[1] 王丹丹[1,2] 景莉君 贾晓斌[1] 

机构地区：[1]中国中医科学院江苏分院、江苏省中医药研究院、国家中医药管理局中药口服释药系统重点研究室,江苏南京210028 [2]江苏大学药学院,江苏镇江212013

出　　处：《中草药》2014年第14期2036-2041,共6页Chinese Traditional and Herbal Drugs

基　　金：国家自然科学基金资助项目(30973978,81274088);江苏省自然科学基金资助项目(BK2011866,BK20141507,BK2011866);江苏省第十批“六大人才高峰”(2013-YY006)

摘　　要：目的利用泼尼松龙诱导的斑马鱼骨质疏松模型筛选朝鲜淫羊藿抗骨质疏松活性部位,探讨斑马鱼骨质疏松模型的适用性与合理性。方法将受精后4 d的斑马鱼幼鱼暴露在25μmol/L泼尼松龙模型药物组,含25μmol/L泼尼松龙的15μg/mL依替膦酸二钠阳性药物组,0.5%DMSO溶媒对照组、含25μmol/L泼尼松龙的朝鲜淫羊藿总提物(生药0.1、1.0、10.0、100.0μg/mL)及其大孔树脂水及30%、50%、70%、90%乙醇洗脱部位(生药1.0、10.0、100.0、1 000.0μg/mL)溶液组,28.5℃下在24孔板中培养,每天换液至第9天处死。采用茜素红对各组斑马鱼幼鱼骨骼染色,并以显微检测、数码成像方法定量分析骨骼染色区域。结果与对照组相比,模型组斑马鱼头部骨骼染色面积和染色光密度值显著减少;与模型组比较,除朝鲜淫羊藿大孔树脂水洗脱部位外,朝鲜淫羊藿总提物及30%、50%、70%、90%乙醇洗脱部位的斑马鱼头部骨骼染色面积和染色光密度值显著增加。色谱分析表明30%、50%、70%、90%乙醇分离部位主要含黄酮类成分和少量非黄酮类化合物。结论斑马鱼骨质疏松模型成功用于高效筛选朝鲜淫羊藿的抗骨质疏松活性部位,具简单、高效及可操作性强的优势。Objective Prednisolone-induced osteoporosis model using zebrafish was used to screen the antiosteoporotic active parts of Epimedium koreanum, and to investigate the applicability and rationality of the zebrafish model of osteoporosis. Methods Zebrafish larva at 4 d post fertilization were exposed with 25 μmol/L prednisolone （model group）, 15 μg/mL etidronate disodium with 25 μmol/L prednisolone （positive group）, 0.5% DMSO （vehicle control group）, E. koreanum extracts （crude drug 0.1, 1.0, 10.0, and 100.0 μg/mL） with 25 pmol/L prednisolone, and their water and 30%, 50%, 70%, and 90% ethanol elution parts of macroporous resin （crude drug 1.0, 10.0, 100.0, and 1 000.0 μg/mL）. All groups were incubated in 24-well plates （28.5 ℃） for 9 d until execution, and then zebrafish skeleton was anesthetized and fixed for staining with alizarin red. Quantitative analysis of the stained area was performed by microscopic inspection and digital imaging methods to reflect the amount of zebrafish head skeleton mineralization. Results The head skeleton mineral area and integrated optical density （IOD） of 25 μmol/L prednisolone model group were significantly decreased when compared with vehicle control group, and E. koreanum extract and its 30%, 50%, 70%, and 90% ethanol elution parts of macroporous resin could rescue the further bone loss of zebrafish induced by prednisolone when compared with model group. HPLC analysis indicated that the components of 30%, 50%, 70%, and 90% ethanol elution parts ofmacroporous resin mainly included flavonoids and minor nonflavonoids components. Conclusion This novel osteoporosis zebrafish model is successfully used to screen the antiosteoporotic active parts ofE. koreanum, which has the advantages of simplity, high efficiency, and easy to perform.

关 键 词：斑马鱼 骨质疏松 朝鲜淫羊藿 活性部位 筛选 

分 类 号：R285.5[医药卫生—中药学]