过度喂养建立斑马鱼幼鱼肥胖模型  

Establishment of a diet-induced obesity model in zebrafish larvae

作　　者：郑新春[1] 刘莉[1] 戴文聪[1] 王坤元[1] 陈小辉[2] 赵灵凤[2] 黄志斌[2] 侯金林[1] 

机构地区：[1]器官衰竭防治国家重点实验室//广东省病毒性肝炎研究重点实验室//南方医科大学南方医院感染内科 [2]南方医科大学基础医学院细胞生物学教研室,广东广州510515

出　　处：《南方医科大学学报》2016年第1期20-25,共6页Journal of Southern Medical University

基　　金：国家自然科学基金基金(81372283)~~

摘　　要：目的探讨利用过度喂养的方法建立斑马鱼幼鱼肥胖模型。方法将7 dpf大小幼鱼随机分为30 mg/d正常喂养组和180 mg/d过度喂养组,每组100条,喂养20 d。分别测量每组幼鱼体质量、体长、BMI、TG和TCH含量,整体油红O染色、冰冻切片油红染色和HE染色观察幼鱼肝脏病理改变。尼罗红染色活体观察幼鱼肝内脂滴和体内脂肪组织动态变化。结果过度喂养组幼鱼体质量、体长、BMI和TG含量比与正常喂养组明显增加。整体油红O染色和病理学结果显示过度喂养幼鱼肝脏脂肪变性率(89.4%)比正常喂养(20.7%)明显高,肝脏出现大泡性脂肪变性。尼罗红染色观察到过度喂养幼鱼肝内脂滴聚集;随着喂养时间延长脂肪组织聚集逐渐增多且分布部位增加。饥饿状态下幼鱼脂肪和肝内脂滴被动员消耗而重新喂养后脂肪组织重新被诱导。结论用过度喂养方法成功建立斑马鱼幼鱼肥胖模型,尼罗红染色为活体观察幼鱼脂肪和肝内脂质提供很好的实验方法,为研究肥胖提供实验模型。Objective To establish a diet-induced obesity model in zebrafish larvae. Methods At 7 days post-fertilization（dpf）,200 zebrafish larvae with normal development were randomly allocated to two groups with the feeding quantity of 30 mg per day（normal feeding group） or 180 mg per day（overfed group） for 20 days. The weight, length, BMI, triglyceride（TG） and total cholesterol（TCH） of each group were measured. Whole- mount Oil Red O staining, frozen Oil Red O staining and hematoxylin- eosin（HE） staining were used to estimate the rate of hepatic steatosis and liver histology of the zebrafish. The dynamic change of hepatic lipid droplets and distribution of adipose tissue were observed with Nile Red staining in overfed zebrafish in vivo. Results The weight, length, BMI and TG of overfed zebrafish were significantly increased compared with those in normal feeding group. Whole-mount Oil Red O staining showed that the percent of hepatic steatosis in overfed group（89.4%） was markedly higher than that in normal feeding group（20.7%）. Macrovesicular steatosis was observed in the liver of the overfed larvae. Nile Red staining visualized hepatic lipid droplets and the distribution of larval adipose tissue, which increased with feeding time in the overfed zebrafish. Starving larvae showed depletion of fat and hepatic lipid, and adipose tissue was induced after refeeding. Conclusions We successfully established an diet-induced obesity model in zebrafish larva,in which Nile Red staining allows in vivo observation of the adipocytes and hepatic lipid droplets.

关 键 词：斑马鱼 肥胖 肝脂肪变性 尼罗红 活体成像 

分 类 号：R723.14[医药卫生—儿科] R-332[医药卫生—临床医学]