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。
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.
出处
《南方医科大学学报》
CAS
CSCD
北大核心
2014年第6期777-782,共6页
Journal of Southern Medical University
基金
国家科技重大专项(2012ZX10002003)~~