大鼠肝纤维化模型中肝细胞凋亡及其调控基因的表达  

Hepatocellular apoptosis and its apoptosis-regulating gene in rat liver fibrosis model

作　　者：吕鹏[1] 罗和生[1] 余保平[1] 

机构地区：[1]湖北医科大学附属第一医院消化内科,湖北省武汉市430060

出　　处：《世界华人消化杂志》2001年第2期165-169,共5页World Chinese Journal of Digestology

摘　　要：目的观察肝细胞凋亡及其调控基因在肝纤维化形成过程中的动态变化,探讨其在肝纤维化发病机制中的作用。方法雄性Wistar大鼠28只(体质量85g～95g)随机分为4组。A,B,C为实验组,采用腹腔注射CCl_4的方法造模;D为对照组,代之以生理盐水。于2,4,6wk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HE染色、细胞凋亡原位TUNEL检测、BAX,BCL-X_L蛋白S-P法免疫组化染色。结果 HE染色A,B,C三组分别符合肝纤维化的Ⅰ,Ⅲ,Ⅳ期,凋亡的肝细胞在肝组织中散在分布TUNEL检测各实验组(A,B,C)肝细胞凋亡级数均高于对照组(D)(2.5±0.6),(2.8±0.4),(2.0±0.6)vs(0.3±0.5),(P<0.01,q_(AD)=9.86,q_(BD)=11.36,q_(CD)=7.59).S-P法免疫组化检测BAX,BCL-X_L主要分布于中央静脉及肝细胞坏死灶周围,在各实验组(A,B,C)中的表达强度均高于对照组(D)。BAX:(1.0±0.6),(1.5±0.6),(1.2±0.4)vs(0.3±0.5),(P<0.05,q_(AD)=3.05,q_(CD)=3.82,A,CvsD;P<0.01q_(BD)=5.32,BvsD)。BCL-X_L:(1.5±0.6),(1.3±0.8),(1.8±0.4)vs(0.3±0.5),(P<0.01,q_(AD)=4.88,q_(BD)=4.16,q_(CD)=6.25,A,B,CvsD)。各实验组的BAX/BCL-X_L高于对照组((0.9+0.8),(2.2±1.2),(0.6±0.4)vs(0.4±0.2),(P<0.05,q_(AD)3.44,q_(BD)=3.40,A,CvsD;P<0.01,q_(BD)=5.14,BvsD)。凋亡级数与BAX/BCL-X_L呈正相关(r=0.60137,P<0.05)。结论肝细胞凋亡在肝纤维化发生过程中存在动态变化,凋亡调控基因bax,bcl_x_1的表达及其比率是变化的主要原因之一。AIM To study the effect of hepatocellular apoptosis andapoptosis-regulating gene on the pathogenesis of liver fibrosis. METHODS Twenty-eight male Wistar rats were randomly divided into 4 groups. A, B, C were experimental groups, while D was normal control group. The rats of experimental groups were intraperitoneally injected with CCl_4, and the normal control with normal saline. The rats of group A, B, C were killed after 2, 4, 6 weeks respectively, and those of the normal control were killed at the same time. The livers were taken for pathologic examination, in situ TUNEL and immunohistochemical stain of BAX and BCL-X_L by streptavidin peroxidase method. RESULTS The expression of hepatocellular apoptosis in experimental groups was significantly higher than in normal control group (2.5±0.6), (2.8±0.4), (2.0±0.6) vs(0.3±0.5), (P<0.01, q_Ao=9.86, q_BD=11.36, q_CD=7.59, A, B, CvsD). The positive rates of BAX and BCL-X_Lwere also higher than in control. BAX:(1.0±0.6), (1.5±0.6), (1.2±0.4) vs 10.3±0.5), (P<0.05, q_AD=3.05,q_CD=3.82, A, CvsD; P<0.01 q_BD=5.32, BvsD). BCL-X_L:(1.5±0.6), (1.3±1.8), (1.8±0.4) vs (0.3±0.5), (P<0.01, q_AD=4.88, q_BD=4.16, q_CD=6.25, A, B, CvsD). BAX/ BCL-X_L of experimental groups was significantly higher than that of normal control group (0.9±0.8), (2.2±1.2), (0.6+0.4) vs (0.4±0.2), (P<0.05, q_AD=3.44, q_CD=3.40, A, CvsD; P<0.01, q_BD=5.14, BvsD). The rates of hepatocellular apoptosis and BAX/BCL-X_L were positively correlated (r=0.6037, P< 0.05). CONCLUSION There were dynamic changes of hepatocellular apoptosis in the development of liver fibrosis. The changes in apoptosis-regulating gene expression and their ratios may be one of the major reasons.

关 键 词：肝硬化 病理学 脱噬作用 基因表达调控 

分 类 号：R575.2[医药卫生—消化系统]