大鼠肝细胞凋亡和坏死动物模型的建立及评价

Establishment and Evaluation of the Animal Models of Hepatic Apoptosis and Necrosis in Rats

目的:通过大鼠肝右叶血供干预分别建立肝细胞凋亡和坏死动物模型,为进一步的磁共振成像实验奠定基础。方法:选取体重为250-300克的SD大鼠90只,随机分为A、B、C三组,每组30只,分别行肝右叶门静脉和肝动脉结扎术、门静脉结扎术及假手术,各组在术后3 h及1、3、7和14 d分别处死6只大鼠,取肝脏标本送病理及电镜检查。结果: A组30只大鼠肝右叶发生凝固性坏死,光镜及电镜下均呈典型的坏死改变,TUNEL染色呈阴性;B组30只大鼠肝右叶发生小灶性凋亡,光镜下可见肝细胞核固缩改变,电镜下见肝组织核边集和凋亡小体,TUNEL染色呈阳性改变。术后3 h 即可见凋亡细胞,24 h时凋亡细胞数最多,7 d后肝组织开始发生小灶性坏死;C组30只大鼠肝右叶未发生凋亡和坏死的病理改变。结论:大鼠肝右叶门静脉和肝动脉双结扎可建立胆脏凝固性坏死模型,单纯肝右叶门静脉结扎能构建肝细胞小灶性凋亡模型。

Objective：To establish rat models of hepatic cell apoptosis and necrosis by intervention of blood supply of the right hepatic lobe. Methods： 90 SD rats with body weight 250 - 300g were randomly divided into three groups, with 30 rats in each group. Group A had ligation of both the right hepatic artery and portal vein for the aim of necrosis. Group B had ligation of the right portal vein for apoptosis. Group C had no intervention of blood supply for control and comparison. Six rats from each group were sacrificed at 3 h, 1d, 3d, 7d and 14d after operation,and the right hepatic lobes were sampled and examined using light microscopy and electronic microscopy. Results： All of the rats from group A had typical coagulative necrosis in right hepatic lobe under light microscopy and electron microscopy,and were negative on TUNEL staining. In group B rats,focal apoptosis in right hepatic lobe could be revealed. There were pyknosis under light microscope; karyon bound gathering and apoptosis small body under electron microscopy. Results of TUNEL staining were positive. In the right lobe,apoptosis could be seen at 3h,with a peak at 24h;focal necrosis accompanied apoptosis could be revealed at 7d after surgery. All thirty rats from group C showed no signs of necrosis and apoptosis in the right hepatic lobe. Conclusion： Right hepatic lobe coagulative necrosis can be obtained in rat models by ligation of right portal vein and hepatic artery, focal apoptosis can be obtained simply by right portal vein ligation in rat models.