摘要
目的：采用复合因素建立大鼠肝纤维化模型,并对其肝脏纤维化病变和相关指标进行动态观察。方法：雄性Wistar大鼠随机分为对照组和肝纤维化模型组,模型组大鼠每周2次乙醇灌胃、猪血清腹腔注射、CCl4葵花仔油溶液皮下注射,同时每日喂饲高脂饲料进行联合造模。分别在造模后第2、4、6、7、8、9、10周检测各组大鼠血清丙氨酸转氨酶（ALT）、天冬氨酸转氨酶（AST）、总蛋白（TP）、白蛋白（Alb）、透明质酸（HA）、层粘连蛋白（LN）、转化生长因子β1（TGF-β1）、基质金属蛋白酶（MMP-2、MMP-9、MMP-13）及其抑制物（TIMP1、TIMP-2）水平,并观察肝脏病理病变。结果：从第2周起,模型组大鼠血清ALT、AST水平开始升高,各时间点与对照组比较均具有显著性差别;血清HA和LN水平逐渐增高,第7～10周与对照组比较均具有显著性差别;血清MMP-2水平逐步升高,第4、6～10周与对照组相比均具有显著性差别;血清MMP-13水平从第6周起逐渐降低,第9和10周时显著低于对照组;血清TIMP-1和TIMP-2水平随造模时间延长逐步升高,TIMP-1在第4,6～10周与对照组相比均具有显著性差别,TIMP-2在第7～10周显著高于对照组;模型组大鼠各时间点血清TGF-β1水平均显著高于对照组。病理组织学检查显示,第6周时,模型组大鼠肝脏细胞广泛性空泡变性,汇管区成纤维细胞增生,肝小叶间隔可见胶原纤维沉积,出现典型的肝纤维化改变;到第8周时,形成典型的假小叶,肝硬化形成。结论：采用酒精灌胃,猪血清腹腔注射,喂饲高脂饲料,并结合CCl4皮下注射6周后,可成功建立大鼠肝纤维化模型;HA、LN、TGF-β1、MMP-2、TIMP-1和TIMP-2等可作为较好反映大鼠肝纤维化病变的无创伤化血清学指标。
Objective：To establish the model of liver fibrosis（ALF） in rats induced by complex factors,and observe dynamicly its hepatic fibrosis lesions.Methods：54 healthy male rats were divided randomly into two groups：control group（n=9） and liver fibrosis group（n=45）.The rats in the liver fibrosis group were given intraperitoneal injection of swine serum,subcutaneous injection of CC14 and gastric perfusion alcoho twice a week,and were fed with high fat food daily to induce ALF.During induction process,the serum levels of alanine aminotransferase（ALT）,aspartate aminotransferase（AST）,total protein（TP）,albumin（Alb）,Hyaluronic acid（HA）,laminin（LN）,transforming growth factor beta1（TGF-β1）,matrix metalloproteinases（MMP-2,MMP-9,MMP-13）,matrix metalloproteinases inhibitory factors（TIMP-1,TIMP-2） and histopathological changes of liver tissue were observed at the 2th,4th,6~10th week repectively.Results：From the beginning of the 2nd week,the ALT and AST level in serum of the model group went up,and ALT,AST level had significant differences compared with those in control group respectively at each time point;the serum levels of HA and LN group went up gradually,At the 7~10th week,compare with control group,there had significant differences;the serum levels of MMP-2 went up gradually,At the 4 th week and 6~10th week,compare with control group,there had significant difference;the serum levels of MMP-13 went down gradually from the 6th week,and was lower than that of control group at the 9th and 10th week;the serum levels of TIMP-1,TIMP-2 went up gradually from the 2th week,and At the 4th week and 6~10th week,compare with control group,there had significant difference for TIMP-1,TIMP-2 was higher than that of control group at the 7~10th week;the serum levels of TGF-β1 of model group was higher than that of control group at each time point.The histopathological changes of liver tissue shows：the hepatocyte of the rats in the liver fibrosis group were vacuolar degeneration widely,Fibroblast hyperplasia were founded between portaltracts.Collagen fibers were founded between hepatic lobules.That is typical change of ALF.At the 8th week,pseudolobuli were founded,cirrhosis was formed.Conclusion：It is a successful way to induce ALF in rats by complex factors including alcohol,swine serum,high fat diet and CC14 for 6 weeks.Moreover,HA,LN,TGF-β1,MMP-2,TIMP-1 and TIMP-2 in serum can reflect preferablely hepatic fibrosis lesions of rat as good targets.
出处
《中国卫生检验杂志》
CAS
2010年第3期547-550,共4页
Chinese Journal of Health Laboratory Technology
基金
浙江省科技厅实验动物科技计划(2008F80013)
