uPA-/-小鼠肝纤维化模型中的α-SMA、MMP13表达分析与模型评价

doi:10.3969/j.issn.1674-5817.2012.06.008

实验动物与比较医学 ›› 2012, Vol. 32 ›› Issue (6): 493-498.DOI: 10.3969/j.issn.1674-5817.2012.06.008

uPA^-/-小鼠肝纤维化模型中的α-SMA、MMP13表达分析与模型评价

夏敏杰¹, 成倩倩¹, 王玉柱¹, 田芳¹, 温成丽¹, 王晓东^2,3, 李卫华¹, 丁训诚¹

1.上海市计划生育科学研究所, 上海 200032;
2.上海实验动物研究中心, 上海 201203;
3.上海西普尔-必凯实验动物有限公司, 上海 201203

收稿日期:2012-02-23 出版日期:2012-12-25 发布日期:2012-12-25
作者简介:夏敏杰(1983),硕士,研究实习员;E-mail:minjie.xia@yahoo.com.cn
基金资助:
上海市科委基金项目(09140902800)

The Expression of α-SMA, MMP13 and Model Evaluation in uPA Knock-out (uPA^-/-) Mice with Liver Fibrosis

XIA Min-jie¹, CHENG Qian-qian¹, WANG Yu-zhu¹, TIAN Fang¹, WEN Cheng-li¹, WANG Xiao-dong^2,3, LI Wei-hua¹, DING Xun-cheng¹

1. Shanghai Institute of Planned Parenthood Research, Shanghai 200032, China;
2. Shanghai Lab oratory Animal Research Center, Shanghai 201203, China;
3. Sino-British SIPPR/BK Laboratory. Animal LTD., CO, Shanghai 201203, China

Received:2012-02-23 Online:2012-12-25 Published:2012-12-25

摘要/Abstract

摘要： 目的采用uPA基因缺失(uPA^-/-)小鼠构建肝纤维化动物模型,经α-平滑肌肌动蛋白(α-SMA)与金属基质蛋白酶-13(MMP13)的表达分析,评价其可行性。方法选取成年雄性BL/6J野生型和uPA^-/-基因缺失型小鼠, 分为4组: 对照组(Con-WT, Con-uPA^-/-)和模型组(Mod-WT, Mod-uPA^-/-),每组20只。模型组小鼠腹腔注射10% CCl₄, 对照组小鼠腹腔注射橄榄油, 每周2次, 连续6周, 诱导小鼠肝纤维化。用Real-time PCR方法检测小鼠肝脏组织中α-SMA和MMP13的mRNA表达, Western blot及免疫组化分析α-SMA和MMP13的蛋白表达。结果 Real-time PCR结果显示, uPA^-/-小鼠的肝脏中α-SMA、MMP13的mRNA表达量显著高于WT小鼠; Western blot结果显示, α-SMA蛋白和MMP13蛋白在对照组小鼠肝脏中几乎不表达, 在Mod-WT中有少量表达,在Mod-uPA^-/-中表达明显增多; 免疫组化结果显示, Mod- uPA^-/-的α-SMA和MMP13表达高于Mod-WT。结论 α-SMA的mRNA和蛋白表达在uPA^-/-小鼠中均明显升高, uPA基因的缺失促进了肝星型细胞向肌成纤维细胞的转化,从而加速了细胞外基质蛋白在肝脏中的沉积; MMP13蛋白表达在肝纤维化模型小鼠中均有明显升高。uPA基因缺失(uPA^-/-)小鼠是建立肝纤维化模型的易感品系。

关键词: uPA基因缺失小鼠, 肝纤维化, α-SMA, MMP13

Abstract: Objective To evaluate the feasibility of uPA knock-out (uPA^-/-) mice liver fibrosis model by analysing the expression of α-Smooth Muscle Actin (α-SMA) and Matrix Metalloproteinase-13 (MMP13). 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 (Con-WT, Con- uPA^-/-) and liver fibrosis model groups (Mod-WT, Mod-uPA^-/-). Mice were intraperitoneally injected with 0.15ml 10% CCl₄ (or olive oil as control) twice per week for 6 weeks to induce liver fibrosis. The mRNA expression of á-SMA and MMP13 was detected by real-time polymerase chain reaction (PCR) and the protein expression of α-SMA and MMP13 was analyzed by western blot and immunohistochemistry. Result The real-time PCR showed that the mRNA expression of α-SMA, MMP13 in uPA^-/- mice wes significantly higher than that in WT mice. The western blot results indicated that there were no expression of α-SMA protein and MMP13 protein of the mice liver in the control group, a few expression of α-SMA protein in Mod-WT group, and a significant increase of α-SMA protein expression in the Mod-uPA^-/- group. The immunohistochemistry results demonstrated that the protein expression of α-SMA and MMP13 of Mod-uPA^-/- group was higher than Mod-WT group. Conclusion The mRNA and protein expression of α-SMA were significantly higher in the uPA^-/- mice. The knocking-out of the uPA gene promoted the hepatic stellate cells transforming into myofibroblasts, thus speeding up the deposition of extracellular matrixin in the liver. Also, the protein expression of MMP13 in liver fibrosis model mice was significantly increased. uPA^-/-mice are susceptible animal strain to establish liver fibrosis model.

Key words: uPA knock-out mice, Liver fibrosis, α-SMA, MMP13

中图分类号:

Q95-33

夏敏杰, 成倩倩, 王玉柱, 田芳, 温成丽, 王晓东, 李卫华, 丁训诚. uPA^-/-小鼠肝纤维化模型中的α-SMA、MMP13表达分析与模型评价[J]. 实验动物与比较医学, 2012, 32(6): 493-498.

XIA Min-jie, CHENG Qian-qian, WANG Yu-zhu, TIAN Fang, WEN Cheng-li, WANG Xiao-dong, LI Wei-hua, DING Xun-cheng. The Expression of α-SMA, MMP13 and Model Evaluation in uPA Knock-out (uPA^-/-) Mice with Liver Fibrosis[J]. Laboratory Animal and Comparative Medicine, 2012, 32(6): 493-498.

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uPA^-/-小鼠肝纤维化模型中的α-SMA、MMP13表达分析与模型评价

The Expression of α-SMA, MMP13 and Model Evaluation in uPA Knock-out (uPA^-/-) Mice with Liver Fibrosis

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uPA-/-小鼠肝纤维化模型中的α-SMA、MMP13表达分析与模型评价

The Expression of α-SMA, MMP13 and Model Evaluation in uPA Knock-out (uPA-/-) Mice with Liver Fibrosis

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uPA^-/-小鼠肝纤维化模型中的α-SMA、MMP13表达分析与模型评价

The Expression of α-SMA, MMP13 and Model Evaluation in uPA Knock-out (uPA^-/-) Mice with Liver Fibrosis