大鼠肝纤维化模型肝细胞凋亡及肝细胞坏死的动态分析

Dynamic analysis of hepatocellular apoptosis and necrosis in liver fibrosis model of rat

目的 观察肝细胞凋亡和肝细胞坏死在肝纤维化形成过程中的变化 ,探讨其在肝纤维化发病机制中的作用。方法 雄性Wistar大鼠 32只 ( 85～ 95克 )随机分为 4组。A ,B ,C为实验组 ,采用腹腔注射CCl4 的方法造模 ;D为对照组 ,代之以生理盐水。于 2 ,4,6周末分别处死A ,B ,C三组大鼠 ,D组同期处理。取肝脏石蜡包埋 ,连续切片做病理学检查、细胞凋亡原位检测。测定血清ALT。结果 病理学检查A ,B ,C ,D四组分别符合肝纤维化的S1,S3 ,S4 ,S0 期。各实验组 (A ,B ,C)的肝细胞坏死记分值均高于对照组 (D) ( 1 6 7± 0 82 ) ,( 2 83± 0 75 ) ,( 2 5 0± 1 0 5 )vs( 0 16± 0 4 1) ,(P <0 0 1) ,肝细胞凋亡级数也均高于对照组 (D) ( 2 5 0± 0 5 5 ) ,( 2 83± 0 4 1) ,( 2 0 0± 0 6 3)vs( 0 33± 0 5 2 ) ,(P <0 0 1)。各实验组 (A ,B ,C)的ALT均高于对照组 (D) ( 76 83± 2 1 87) ,( 117 0 0±15 94) ,( 10 3 6 7± 36 6 8)vs( 39 16± 2 32 ) ,(P <0 0 5 )。结论 肝细胞凋亡和肝细胞坏死在肝纤维化发生过程中存在动态变化 ,二者在其不同时期发挥的作用不同。

Aim To study the effects of hepatocellular apoptosis and necrosis on the pathogenesis of liver fibrosis.Methods 32 male Wistar rats were randomly divided into 4 groups.A,B,C were experimental groups,while D was control group.The rats of experimental groups were intraperitoneally injected with CCl 4,and the control with normal saline.The rats of group A,B,C were killed after 2,4,6 weeks respectively,and those of the control were killed at the same time as a comparison.The livers were taken for pathologic examination,in situ terminal deoxynucleatidy1 transferase-mediated deoxyuridine triphosphate nick end labeling (TUNEL).The serum ALT was tested too.Results The rats of group A,B,C,D respectively corresponded to stage S 1,S 3,S 4,S 0 of the liver fibrosis.The score of hepatocellular apoptosis in experimental groups were significantly higher than control group (2.50±0.55),(2.83±0.41),(2.00±0.63)vs(0.33±0.52),(P<0.01),and so were the hepatocellular necrosis score(1.67±0.82),(2.83±0.75),(2.50±1.05)vs(0.16±0.41),(P<0.01).In addition,serum ALT in experimental groups were significantly higher than control group (76.83±21.87),(117.00±15.94),(103.67±36.68)vs(39.16±2.32),(P<0.05).Conclusions There are dynamic changes of hepatocellular apoptosis and necrosis in the development of liver fibrosis.They play different roles in different stages.