接骨汤抗泼尼松龙诱导斑马鱼骨丢失的作用研究  

Preventive Effect of Jie-Gu-Tang on Osteoporosis Model Using Zebrafish Induced by Prednisolone

作　　者：景莉君 王长梅[1,2] 韦英杰[1,2,3] 宁青[1,2] 谢林[1,2,3] 

机构地区：[1]中国中医科学院江苏分院 [2]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,南京210028 [3]南京中医药大学附属江苏省中西医结合医院,南京210028

出　　处：《世界科学技术-中医药现代化》2014年第5期1071-1075,共5页Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology

基　　金：江苏省科技厅自然科学基金(BK2011866):模式生物斑马鱼壮骨效应评价模型的建立与应用研究,负责人:韦英杰;国家自然科学基金委面上项目(30973978):基于模式生物斑马鱼模型的中药代谢研究新方法的建立,负责人:韦英杰;江苏省委组织部“六大人才高峰”(2013-YY-006):以二维斑马鱼模型联合色谱联用技术高效筛选中药抗骨质疏体内外活性成分,负责人:韦英杰;江苏省委组织部“六大人才高峰”(2010-WS-071):基于“肝主筋”组方通过TNF-α/NF-κB通路干预盘源性颈痛,负责人:谢林;江苏省中医药局中医药领军人才项目(CJ200916):补肾益气活血方多靶点阻断盘源性下腰痛发生发展机制研究,负责人:谢林

摘　　要：目的:本文采用泼尼松龙诱导的斑马鱼骨质疏松模型,研究临床优效方接骨汤防止骨丢失,促进骨生成的作用。方法:将受精后5天的斑马鱼幼鱼暴露在含25μmol·L-1泼尼松龙的不同浓度接骨汤(0.025、0.25、2.5、25、100 mg生药·L-1)溶液中,同时设25μmol·L-1泼尼松龙模型药物组,含25μmol·L-1泼尼松龙的依替膦酸二钠(15、30 mg·L-1)阳性药物组及0.5%DMSO溶媒对照组。28.5°C下在24孔板中培养,每天换液培养至第10天处死。采用茜素红对培养10天的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:与溶媒阴性对照组相比,25μmol·L-1浓度泼尼松龙组(模型组)的斑马鱼骨骼染色面积和染色光密度值显著减少;与模型组比,依替膦酸二钠(15、30 mg·L-1)和接骨汤(2.5、25、100 mg生药·L-1)组的鱼骨染色矿化面积和累积光密度值显著增加,且呈一定量效关系。结论:用泼尼松龙诱导的斑马鱼骨质疏松模型成功评价了接骨汤防止骨丢失,促进骨生成的作用,为该模型的合理性与可靠性提供依据。Prednisolone-induced zebrafish osteoporosis model was used to explore the bone-strengthening effect of Jie-Gu-Tang （JGT）. Zebrafish larvae of 5 days post fertilization （d.p.f.） were co-exposed with 25 p, mol .L^-1 pred- nisolone and a series of JGT solutions with a range of concentrations （0.025, 0.25, 2.5, 25 and 100 mg crude herbper liter）. The 25 μmol. L-I prednisolone was selected as the model group. Etidronate disodium （15 and 30 mg. mL^-1） with 25 μmol-L^-1 prednisolone was used as the positive group. And 0.5% DMSO was used as the vehicle control group. All groups were incubated in 24-well plates （28.5℃） until 10 d.p.f. Zebrafish skeleton at 10 d.p.f, was anes- thetized and fixed for staining with alizarin red. Quantitative analysis of the stained area was performed by microscop- ic inspection and digital imaging methods to reflect the amount of zebrafish head skeleton mineralization. The results showed that prednisolone group at 25 μmol.L^-1 concentration can obviously decrease the staining area and the stain- ing optical density values when compared with the vehicle control group （0.5% DMSO）. Compared with the model group, both etidronate disodium （15 and 30 mg.mL^-1） and JGT （2.5, 25 and 100 mg crude herb per liter） can in- crease the mineralized matrix and integrated optical density （IOD） of zebrafish head skeleton significantly with dose- effect relationship. It was concluded that zebrafish osteoporosis model was successfully used in the evaluation on bone loss prevention and bone formation promotion of JGT, which provided basis for the reliability and reasonability of zebrafish model.

关 键 词：斑马鱼 泼尼松龙 接骨汤 壮骨效应 

分 类 号：R285.5[医药卫生—中药学]