油红O染色在斑马鱼高脂血症模型中的应用  

Application of oil red O staining in zebrafish hyperlipidemia model

作　　者：郑毓倩 张雅迪 施雯婷 张祥 陈昕琳[1,2] ZHENG Yuqian;ZHANG Yadi;SHI Wenting;ZHANG Xiang;CHEN Xinlin(Longhua Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 200032,China;Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China)

机构地区：[1]上海中医药大学附属龙华医院,上海200032 [2]上海中医药大学,上海201203

出　　处：《上海中医药大学学报》2025年第1期12-19,共8页Academic Journal of Shanghai University of Traditional Chinese Medicine

基　　金：国家自然科学基金资助项目(82074371)。

摘　　要：目的:研究确定斑马鱼幼鱼油红O染色(ORO)的最佳条件,并探究其高脂血症(HLP)的最佳造模条件。方法:选取发育正常的4天大(dpf)斑马鱼进行油红O染色,采用正交设计,实验参数为染色时间、油红O浓度、洗脱时间,评价指标为斑马鱼尾部的染色面积比值以及斑马鱼腹部红色光密度(OD);选取发育正常的6 dpf的斑马鱼给予不同浓度蛋黄粉制备高脂血症模型,以第7~15脊间血管油红面积占比为主要评价指标,以总胆固醇(TC)、总三酰甘油(TG)为辅助评价指标。结果:正交分析结果表明油红O浓度对染色结果有影响(P<0.05),极差分析显示0.25%的油红O染色4 h、洗脱2 min后的染色面积比及OD均高于其他条件组合组;析因分析结果显示蛋黄粉浓度及喂养天数均对斑马鱼高脂血症模型有影响,0.3%蛋黄粉喂养2 d的造模效果最佳,浓度升高及喂养天数增加对斑马鱼尾部血管脂质沉积无明显影响;HLP组TC、TG水平也明显高于正常饮食组(P<0.05)。结论:用0.25%的油红O染色4 h、洗脱2 min是斑马鱼幼鱼的油红O染色最佳条件;斑马鱼幼鱼HLP模型的最佳造模条件为0.3%新鲜蛋黄粉喂养2 d。本实验探讨了斑马鱼幼鱼高脂血症造模方法,所筛选出的快速造模方法适用于降脂药物的筛选。Objective:To investigate and determine the optimal conditions for oil red O staining(ORO)of juvenile zebrafish and explore the optimal modeling conditions for juvenile zebrafish hyperlipidemia(HLP).Methods:4 days post fertilization(4dpf)normally developed zebrafish was selected for oil red O staining,and orthogonal design was used.The experimental parameters were staining time,oil red O concentration and elution time.The evaluation indicators were the staining area ratio in zebrafish tail and red optical density(OD)of zebrafish abdomen.6dpf normally developed zebrafish was selected and given egg yolk powder with different concentrations to prepare HLP model,and the proportion of oil red area of the 7th-15th intervertebral vessels was used as the main evaluation index,and total cholesterol(TC)and total triglyceride(TG)were used as auxiliary evaluation indexes.Results:Orthogonal analysis showed that the staining result was affected by the concentration of oil red O(P<0.05),and range analysis showed that the staining area ratio and OD of 0.25%oil red O after 4 h staining and 2 min elution were higher than those of other conditions combination groups.Factorial analysis showed that the zebrafish hyperlipidemia model was affected by the concentration of egg yolk powder and feeding days.The modeling effect by 0.3%egg yolk powder feeding for 2 days was the best,and the lipid deposition in tail vessels of zebrafish was not significantly affected by the increase of concentration and feeding days.The levels of TC and TG in the HLP group were also significantly higher than those in the normal diet group(P<0.05).Conclusions:Staining with 0.25%oil red O for 4 h and elution for 2 min are the optimal conditions for oil red O staining in juvenile zebrafish.The optimal modeling conditions for the high-fat model of juvenile zebrafish were 0.3%fresh egg yolk powder feeding for 2 days.The method of establishing hyperlipidemia model in juvenil zebrafish was explored in this experiment,and the selected rapid modeling method could be us

关 键 词：油红O染色 高脂血症 斑马鱼 正交实验 析因设计 

分 类 号：TS2[轻工技术与工程—食品科学与工程]