uPA基因缺失型小鼠与野生型小鼠肝纤维化模型的比较研究

摘要

目的 分别采用C57BL/6J野生型(WT)和uPA基因缺失(uPA-/-)小鼠构建肝纤维化动物模型,并进行比较研究.方法 选取成年雄性C57BL/6J野生型和uPA-/-小鼠,分为4组:对照组(WT,uPA-/-)和模型组(WT,uPA-/-),每组10只.模型组小鼠腹腔注射10% CCI40.15ml,对照组小鼠腹腔注射橄榄油0.15ml,每周2次,连续4周和6周.测定动物血清中ALT、AST、ALB及A/G等生化指标;盐酸水解法测定肝组织羟脯氨酸(Hyp);用放射免疫法测定血清Ⅲ型前胶原蛋白(PC Ⅲ)含量;用HE染色及Masson三色胶原染色观察肝组织病理学改变.结果 造模4周和6周时,无论是WT模型组小鼠还是uPA-/-模型组小鼠血清ALT和AST活性均较对照组小鼠显著升高,ALB含量降低(P<0.01);造模4周时,仅见uPA-/-小鼠的A/G含量降低(P<0.01).造模4周和6周时,uPA-/-模型组小鼠血清PC Ⅲ水平和肝脏组织Hyp含量较对照组显著升高(P<0.01);造模6周时,WT模型组小鼠才出现血清PC Ⅲ水平和肝脏组织Hyp含量升高(P<0.01),且升高值明显低于uPA-/-模型组小鼠.病理组织学观察显示,uPA-/-模型组的肝脏纤维化程度比WT模型组明显严重.结论 与WT小鼠相比,采用uPA-/-小鼠建立肝纤维化动物模型的造模周期明显缩短,肝纤维化程度更严重.%Objective To build and compare two liver fibrosis models using uPA knock-out (uPA-/-)mice and wild-type (WT) mice. Methods Adult male C57BL/6J WT mice and uPA knock-out (uPA-/-) mice were divided into four groups with 10 mice in each group: control groups (WT, uPA-/-) and liver fibrosis model groups (WT, uPA-/-). Mice were injected intraperitoneally with 0.15ml 10％ CCh (or olive oil as control) 2 times per week for 4 weeks and 6 weeks respectively. Serum alanine aminotransferase (ALT), aspartate(AST), albumin(ALB) and albumin/globulin(A/G) were determined, liver hydroxyproline (Hyp) level was determined using hydrochloric acid hydrolysis method and serum procollagen Ⅲ (PC III) content was measured by radioimmunoassay; liver histopathology were performed using HE staining and Masson staining. Results After modeling for 4 weeks and 6 weeks, serum ALT and AST activity of both WT mice and uPA-/- mice significantly increased while serum Alb content significantly decreased comparing with control (P＜0.01); at 4 weeks, significant decrease of A/G content was found in uPA-/- mice model only (P＜0.01). After modeling for 4 weeks and 6 weeks, serum PC III level and liver Hyp content of uPA-/- mice model significantly increased comparing with control (P＜0.01); At 6 weeks, serum PC III level and liver Hyp content of WT mice models started to increase significantly (P＜0.01), and the elevated values were not as high as those in uPA-/- mice model. Histopathology showed that the degree of liver fibrosis in uPA-/- mice model was more severe than that in WT mice model. Conclusion Compared with C57BL/6J WT mice, establishment of liver fibrosis model by using uPA-/- mice was required shorter time and showed more severe degree of liver fibrosis.