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

Comparative study on Liver Fibrosis Models Using uPA Knock-out Mice and Wild-type Mice

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％CCl40．15ml，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（PCⅢ）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PCⅢ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PCⅢ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.