高胆固醇饮食对斑马鱼脂代谢的影响
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摘要
目的:建立一种稳定可靠的斑马鱼高脂血症模型,并应用不同方法评估高胆固醇饮食对斑马鱼脂代谢的影响。方法:受精5 d后(5 days post fertilization,5dpf)的斑马鱼幼鱼随机分成普通饲料组和高胆固醇饲料模型组,普通饲料为纯蛋黄干粉,高胆固醇饲料是向纯蛋黄干粉中添加不同百分比的胆固醇配制而成,连续喂养10 d后通过显微镜实时观察并对幼鱼生理生化指标进行检测评估。结果:共聚焦显微镜下观察到不同百分比的高胆固醇饮食模型组幼鱼尾部动脉血管脂质沉积量均显著高于普通饲料组(P <0.05),且随着饲料中胆固醇百分比增加,幼鱼动脉血管脂质沉积量也随之增加;增加5%高胆固醇饮食组投喂量,其血管脂质沉积量先显著性增加(P <0.05)后渐趋稳定,但幼鱼生存率逐渐降低(P <0.05);5%高胆固醇饮食组幼鱼体宽明显大于普通饲料组(P <0.01),但体长无明显差异(P> 0.05);5%高胆固醇饮食组幼鱼体内总胆固醇(total cholesterol,TC)和总甘油三酯(total triglyceride,TG)水平较普通饲料组明显升高(P <0.01),且低密度脂蛋白胆固醇(low-density lipoprotein cholesterol,LDL-C)水平也同时升高(P <0.05)。结论:5%高胆固醇饲料喂养10 d可以诱导斑马鱼高脂血症的发生,且适量增加饲料中胆固醇百分比或适当增加高胆固醇投喂量均能加快高脂血症模型建立。
Objective:To establish a stable and reliable model of zebrafish hyperlipidemia and evaluate the effect of high cholesterol diet on the lipid metabolism in zebrafish larvae. Methods:Zebrafish larvae after five days post fertilization(5 dpf)were randomly divided into two groups:normal feed group with dry powder of pure egg yolk,and high cholesterol feed model group with different percentages of cholesterol added to pure egg yolk. After 10 days,the evaluation was carried out by microscopy and detecting the physiological and biochemical indicators in the zebrafish larvae. Results:Confocal microscopy showed that the lipid deposition in caudal artery of juvenile fishes in high cholesterol diet model group was significantly higher than that in common diet group(P < 0.05),and the lipid deposition in caudal artery of juvenile fishes increased with the increase of cholesterol content in diet. When the feed intake of the 5% high cholesterol diet group increased,the lipid deposition in blood vessel increased significantly at the beginning(P <0.05)and subsequently stabilized gradually,but the survival rate of juvenile fishes decreased significantly(P < 0.05). The body width of fish larvae in the 5% high cholesterol diet group was significantly larger than that in common diet group(P < 0.01),but no significant differences in body length were observed between the two groups(P > 0.05). The levels of total cholesterol(TC)and total triglyceride(TG)in juvenile fishes fed with 5% high cholesterol diet were both significantly higher than those fed with normal diet(P <0.01),and the levels of LDL-C were also significantly higher(P <0.05). Conclusion:Feeding 5% high cholesterol diet for 10 days could induce the occurrence of hyperlipidemia in zebrafish. Increasing the percentage of cholesterol in diet or increasing the dosage of high cholesterol properly could accelerate the establishment of hyperlipidemia model.
Objective:To establish a stable and reliable model of zebrafish hyperlipidemia and evaluate the effect of high cholesterol diet on the lipid metabolism in zebrafish larvae. Methods:Zebrafish larvae after five days post fertilization(5 dpf)were randomly divided into two groups:normal feed group with dry powder of pure egg yolk,and high cholesterol feed model group with different percentages of cholesterol added to pure egg yolk. After 10 days,the evaluation was carried out by microscopy and detecting the physiological and biochemical indicators in the zebrafish larvae. Results:Confocal microscopy showed that the lipid deposition in caudal artery of juvenile fishes in high cholesterol diet model group was significantly higher than that in common diet group(P < 0.05),and the lipid deposition in caudal artery of juvenile fishes increased with the increase of cholesterol content in diet. When the feed intake of the 5% high cholesterol diet group increased,the lipid deposition in blood vessel increased significantly at the beginning(P <0.05)and subsequently stabilized gradually,but the survival rate of juvenile fishes decreased significantly(P < 0.05). The body width of fish larvae in the 5% high cholesterol diet group was significantly larger than that in common diet group(P < 0.01),but no significant differences in body length were observed between the two groups(P > 0.05). The levels of total cholesterol(TC)and total triglyceride(TG)in juvenile fishes fed with 5% high cholesterol diet were both significantly higher than those fed with normal diet(P <0.01),and the levels of LDL-C were also significantly higher(P <0.05). Conclusion:Feeding 5% high cholesterol diet for 10 days could induce the occurrence of hyperlipidemia in zebrafish. Increasing the percentage of cholesterol in diet or increasing the dosage of high cholesterol properly could accelerate the establishment of hyperlipidemia model.
引文
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[12]Stoletov K,Fang L,Choi SH,et al.Vascular lipid accumulation,lipoprotein oxidation,and macrophage lipid uptake in hypercholesterolemic zebrafish[J].Circ Res,2009,104(8):952-960
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[14]王泽民.高糖高脂导致的斑马鱼血管病变[D].山东:山东大学医学院,2014
[15]李春启,陈汝家,郭胜亚,等.一种斑马鱼高脂血症模型的建立方法及其应用:CN,CN102907357A[P].2013
[16]李德利,黄梦圆,魏丽圆,等.KLF7b对斑马鱼胚胎发育的影响[J].南京医科大学学报(自然科学版),2018,38(5):605-611
[17]李楠,张昕,史春梅,等.斑马鱼胚胎发育过程中内源性多肽的表达及与眼发育的相关研究[J].南京医科大学学报(自然科学版),2018,38(2):161-1652019-02-01
[2]Egan BM,Li J,White K,et al.2013 ACC/AHA cholesterol guideline and implications for healthy people 2020 cardiovascular disease prevention goals[J].J Am Heart Assoc,2016,5(8):e003558
[3]叶振南,李楠,盛丹丹,等.青钱柳多糖对高脂血症大鼠血脂及抗脂质过氧化作用的影响[J].现代食品科技,2014,30(4):1-5,20
[4]Khan TJ,Ahmed YM,Zamzami MA,et al.Effect of atorvastatin on the gut microbiota of high fat diet-induced hypercholesterolemic rats[J].Sci Rep,2018,8(1):662
[5]Sharma MS,Choudhary PR.Effect of fenugreek seeds powder(Trigonella foenum-graecum L.)on experimental induced hyperlipidemia in rabbits[J].J Diet Suppl,2017,14(1):8
[6]朱磊,路瑛丽,冯连世,等.不同配方高脂饲料构建SD大鼠肥胖模型的实验研究[J].中国运动医学杂志,2016,35(7):642-647
[7]陈剑峰,万勇.不同配方高脂饲料构建高血脂症大鼠模型的比较及评价[J].实验动物科学,2018,35(1):30-34
[8]游晓庆,吴春芳,骆凯,等.高脂膳食诱导新西兰兔高脂血症模型的研究[J].中国保健营养,2016,26(17):414-415
[9]吴韫宏,方显明.家兔实验性高脂血症模型的研究[J].广西中医药大学学报,2004,7(2):1-2
[10]Martín-Castillo A,Castells MT,Adánez G,et al.Effect of atorvastatin and diet on non-alcoholic fatty liver disease activity score in hyperlipidemic chickens[J].Biomed Pharmacother,2010,64(4):275-281
[11]方斌.ApoE基因敲除巴马猪动脉粥样硬化模型的建立及表型分析[D].南京:南京医科大学,2018
[12]Stoletov K,Fang L,Choi SH,et al.Vascular lipid accumulation,lipoprotein oxidation,and macrophage lipid uptake in hypercholesterolemic zebrafish[J].Circ Res,2009,104(8):952-960
[13]韩冰.斑马鱼高血脂模型的建立及小檗胺降血脂机制研究[D].重庆:西南大学,2016
[14]王泽民.高糖高脂导致的斑马鱼血管病变[D].山东:山东大学医学院,2014
[15]李春启,陈汝家,郭胜亚,等.一种斑马鱼高脂血症模型的建立方法及其应用:CN,CN102907357A[P].2013
[16]李德利,黄梦圆,魏丽圆,等.KLF7b对斑马鱼胚胎发育的影响[J].南京医科大学学报(自然科学版),2018,38(5):605-611
[17]李楠,张昕,史春梅,等.斑马鱼胚胎发育过程中内源性多肽的表达及与眼发育的相关研究[J].南京医科大学学报(自然科学版),2018,38(2):161-1652019-02-01