非酒精性脂肪性肝炎及肝纤维化动物模型的建立

Establishment of non-alcoholic steatohepatitis / liver fibrosis model

目的通过胆碱缺乏饮食及乙硫氨酸的联合作用构建非酒精性脂肪性肝炎(non-alcoholic steatohepatitis,NASH)及肝纤维化动物模型。方法选择雄性C57BL/6J小鼠给予含60%高脂的胆碱缺乏饲料,并在第1周的饮水中辅以0.165%的乙硫氨酸加重打击。于第4周、第8周和第16周各随机处死5只小鼠。分离血清,取肝脏组织分别进行HE染色、油红O染色及天狼猩红染色,应用图像分析法对天狼猩红染色结果进行胶原含量半定量分析。结果造模第4周,血清中ALT、AST及TG的含量明显上升,油红O染色显示肝脏组织已经出现大量的脂滴堆积,HE染色可见大量空泡,但无病理性纤维化发生。造模第8周,肝脏组织除可见大量脂滴,天狼猩红染色观察肝组织中央静脉及汇管区周围胶原纤维少量增生。造模第16周,天狼猩红染色在窦周隙内可见成细丝状乃至索状的胶原纤维分布。结论经改良方法后,成功建立小鼠NASH及肝纤维化的模型。

Objective To establish non-alcoholic steatohepatitis （NASH）/liver fibrosis model by the combination of choline-deficient diet and ethionine. Methods Thirty male C57BL/6J mice were randomly divided into control group （n =5） and model group （n = 15）. Model mice were placed on choline-deficient diet （60% kca] fat） and drinking water containing 0. 165% DL-ethionine at the first week. Five mice in model group were killed at 4th, 8th and 16th weeks, respectively. The serums were collected. The histological analysis was conducted by HE staining, Oil Red O staining and Sirius Red staining. The Sirius Red staining was analyzed by image analysis system to semi-quantify the collagen area. Results Compared with control group, liver tissue in model group mice showed markedly increased lipid accumulation but no fibrosis after 4 weeks. Observation of liver tissue revealed a small increasein collagen fiber hyperplasia around the central vein and portal area after 8 weeks. Sirius Red staining revealed filamentous collagen fiber distribution in perisinusoidal space after 16 weeks. Conclusion The mice model of NASH/liver fibrosis can be established successfully by the combination of choline-deficient diet and ethionine.