基于斑马鱼模型对药食同源中药饮品的抗氧化和抗疲劳功效研究

Study on Anti-oxidation and Anti-fatigue Effects of Medicinal and Edible Homologous Beverage for TCM Based on Zebrafish Model

[目的]对2种药食同源中药饮品(香蜂悦颜饮、参杞固元饮)成分进行初步分析,并利用斑马鱼功效模型评价2种饮品的抗氧化和抗疲劳作用。[方法]通过UHPLC-LTQ-Orbitrap-MS液质联用技术,用反相C_(18)色谱柱,采用双压线性离子阱质谱正/负离子双重扫描方式,对样品进行成分分析;对水溶给予甲萘醌方法建立的斑马鱼氧化损伤模型加入梯度浓度(25、50、100μg/mL)的香蜂悦颜饮稀释液干预,同时设置正常组、阳性药组,处理24 h后,Cell ROX避光染色,在荧光显微镜下观察拍照,计算斑马鱼的卵黄囊荧光强度;选择5日龄野生型斑马鱼,加入梯度浓度(25、50、100μg/mL)参杞固元饮稀释液干预,同时设置正常对照组和阳性药对照组,处理18 h后,给药组水溶给予无水亚硫酸钠建立斑马鱼疲劳模型,并利用行为分析仪分析斑马鱼总运动距离。[结果]根据质谱检测数据,香蜂悦颜饮、参杞固元饮分别检测出63和60个化合物。与模型组相比,香蜂悦颜饮干预下的斑马鱼卵黄囊荧光强度显著变弱(P<0.01);与模型对照组相比,参杞固元饮干预下的斑马鱼总运动距离显著更远(P<0.05)。[结论]2种药食同源中药饮品分别具有明确的抗氧化和抗疲劳作用。

[Objective]To analyze the chemical constituents in 2 medicinal and edible homologous beverage(Xiangfeng Yueyan Drink and Shenqi Guyuan decoction),and evaluate the anti-oxidant and anti-fatigue effects.[Method]UHPLC-TQ-Orbitrap-MS mass spectrometer and reverse phase C_(18) column were used to analyze the composition of the samples.The oxidative stress model established by aqueous menadione administration method was added with different gradient concentrations(25,50,100 μg/mL) of Xiangfeng Yueyan Drink dilution.Meanwhile,a normal group and a positive drug control group were set up.After treatment for 24 h,the Cell ROX was stained without light,and the fluorescence intensity of yolk sac was calculated by fluorescence microscope.Select 5-day-old wild type zebrafish and add different gradient concentrations(25,50,100 μg/mL) of Shenqi Guyuan decoction diluent.A normal control group and a positive drug group were set up at the same time.After treatment for 18 h,the fatigue model of zebrafish was established by aqueous solution of anhydrous sodium sulfite in the administration group,and the total movement distance of zebrafish was analyzed by behavior analyzer.[Result]According to the mass spectrometry data,63 compounds were detected in Xiangfeng Yueyan Drink and 60 compounds were detected in Shenqi Guyuan Drink.Compared with the model group,the fluorescence intensity of zebrafish yolk sac significantly weakened under the intervention of Xiangfeng Yueyan Drink(P<0.01);compared with the model control group,the total movement distance of zebrafish under the intervention of Shenqi Guyuan Drink was significantly longer(P<0.05).[Conclusion]The 2 kinds of medicinal and edible homologous beverage have clear anti-oxidation and anti-fatigue effects respectively.