CAJ | 학술논문

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。28.5oC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。
목적：본문채용발니송룡유도적반마어골질소송모형，연구림상우효방접골탕방지골주실，촉진골생성적작용。방법：장수정후5천적반마어유어폭로재함25μmol·L-1발니송룡적불동농도접골탕（0.025、0.25、2.5、25、100 mg생약·L-1）용액중，동시설25μmol·L-1발니송룡모형약물조，함25μmol·L-1발니송룡적의체련산이납（15、30 mg·L-1）양성약물조급0.5% DMSO용매대조조。28.5oC하재24공판중배양，매천환액배양지제10천처사。채용천소홍대배양10천적각조반마어유어골격염색，병이현미검측、수마성상방법정량분석골격염색구역。결과：여용매음성대조조상비，25μmol·L-1농도발니송룡조（모형조）적반마어골격염색면적화염색광밀도치현저감소；여모형조비，의체련산이납（15、30 mg·L-1）화접골탕（2.5、25、100 mg생약·L-1）조적어골염색광화면적화루적광밀도치현저증가，차정일정량효관계。결론：용발니송룡유도적반마어골질소송모형성공평개료접골탕방지골주실，촉진골생성적작용，위해모형적합이성여가고성제공의거。
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 μmol·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 herb per liter). The 25 μmol·L-1 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.