南方医科大学学报 ›› 2014, Vol. 34 ›› Issue (06): 777-.

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二乙基亚硝胺诱导建立斑马鱼肝纤维化模型

王坤元,刘莉,戴文聪,陈小辉,郑新春,侯金林   

  • 出版日期:2014-06-20 发布日期:2014-06-20

Establishment of a hepatic fibrosis model induced by diethylnitrosamine in zebrafish

  • Online:2014-06-20 Published:2014-06-20

摘要: 目的探讨利用二乙基亚硝胺(DEN)致中毒性肝损伤的方法构建斑马鱼肝纤维化模型的可行性。方法3月龄野生型斑马
鱼120条,随机等分为对照组和DEN处理组,分别观察两组斑马鱼生存率及行为学改变,在DEN处理2周、4周和6周末,分别从
两组中随机抽取等量样本测量肝指数,并制作石蜡切片,采用HE染色、Gomori氏网状纤维染色及天狼星红苦味酸染色方法观
察肝纤维化病变。结果DEN处理组斑马鱼在造模期间未见明显行为学改变,与对照组斑马鱼相比,DEN处理组斑马鱼肝指数
在2周、4周和6周末均无统计学差异。病理染色结果显示造模4周末,DEN处理组斑马鱼肝细胞变性坏死,30%斑马鱼肝脏网
状纤维合成分泌增多;6周末,DEN处理组斑马鱼结构紊乱,80%斑马鱼肝脏纤维过度沉积,网状纤维和胶原纤维合成分泌异常
增多,可见纤维结节形成。结论采用DEN诱导中毒性肝损伤方法能够成功地构建稳定的斑马鱼肝纤维模型,为进一步探讨肝
纤维化的发病机制以及药物筛选奠定了实验基础。

Abstract: Objective To establish a zebrafish model of liver fibrosis via diethylnitrosamine (DEN)-induced liver injury.
Methods A total of 120 wild-type 3-month-old zebrafish were randomly divided into DEN-treated group and control group.
The survival rate and behavioral changes of each group were observed. After treatment with DEN for 2, 4, and 6 weeks, liver
index was measured, and liver fibrosis was evaluated with HE staining, Gomori staining and Sirius red staining. Results No
obvious behavioral change was observed in DEN-treated group during the experiment. Compared with that in control group,
the liver index of zebrafish in DEN-treated group showed no significantly changes at the time points of observation.
Proliferation of reticulate fibers was found in 30% of zebrafish treated with DEN for 4 weeks, and the rate increased to 80% at 6
weeks when reticulate fibers and collagen fibers actively proliferated to result in fiber collapse and formation of fibrotic
nodules. Conclusion A stable zebrafish liver fibrosis model was successfully established by inducing liver damage to facilitate
studies of the pathogenesis of liver fibrosis and screening therapeutic drugs.