肝胆胰外科杂志 ›› 2020, Vol. 32 ›› Issue (8): 488-493.doi: 10.11952/j.issn.1007-1954.2020.08.009

• 论著 基础研究 • 上一篇    下一篇

肝细胞特异性HuR基因敲除小鼠模型的表型研究

黄政伟1 ,季晓克1 ,黄文铅1 ,黄思琪2 ,李义孟1 ,吴燚阳3 ,戴克智4 ,张启瑜1   

  1. 1.温州医科大学附属第一医院 肝胆外科,浙江 温州 325015;温州医科大学附属第二医院,浙江 温州 325027,2.超声影像科,3.肝胆外科;4.温州医科大学附属康宁医院 精神卫生研究所,浙江 温州 325007)
  • 收稿日期:2020-05-18 出版日期:2020-08-15 发布日期:2020-09-06
  • 通讯作者: 张启瑜,主任医师,教授,博士生导师,E-mail:qiyuz@126.com。
  • 作者简介:黄政伟(1993-),男,浙江温州人,在读硕士。
  • 基金资助:
    温州市医药卫生科学研究项目计划(2019A17),国家自然科学基金青年项目(81900566)。

Phenotype study of hepatocyte-specific HuR gene knockout in mice model

HUANG Zheng-wei1 , JI  Xiao-ke1 , HUANG Wen-qian1 , HUANG Si-qi2 , LI Yi-meng1 , WU Yi-yang3 , DAI Ke-zhi4 , ZHANG Qi-yu1   

  1. 1Department of Hepatobiliary Surgery, The First Affiliated Hospital of Wenzhou Medical University, Zhejiang 325015, China;2Department of Ultrasound Imaging,3Department of Hepatobiliary Surgery, The Second Affiliated Hospital of Wenzhou Medical University, Zhejiang 325027, China;4Psychic Health Research Institute, The Affiliated Kangning Hospital of Wenzhou Medical University, Zhejiang 325007, China
  • Received:2020-05-18 Online:2020-08-15 Published:2020-09-06

摘要:

目的 探讨肝细胞特异性敲除HuR基因对小鼠肝脏功能的影响。方法 从美国Jackson实验室引进LoxP标记的人抗原R(human antigen R,HuR)基因小鼠(HuRfl/fl)和Alb-Cre转基因小鼠。杂交后,经数代选育配种,建立肝细胞特异性HuR基因敲除小鼠(HuRLKO)模型。PCR技术鉴别小鼠基因型,蛋白免疫印迹和免疫荧光实验检验小鼠肝脏的HuR表达水平,同时HE染色观察肝脏结构变化。高脂喂养HuRfl/fl/Alb-CreHuRfl/fl两组小鼠,观察小鼠体重、肝重和肝体比变化,提取血清检测肝脏损伤和脂肪代谢相关指标。结果 PCR成功检测筛选子代小鼠基因型,包括HuRfl/fl/Alb-CreHuRfl/fl小鼠。蛋白免疫印迹实验和免疫荧光实验证明小鼠肝细胞特异性HuR基因敲除成功。HE染色结果提示HuRLKO小鼠的肝细胞出现变性水肿、局部坏死。高脂喂养实验提示两组小鼠在肝重、肝体比、AST、ALT、HDL-C、血糖上的差异具有统计学意义(P<0.05),但在体重、TG、LDL-C的差异没有统计学意义(P>0.05)。结论 HuR基因是维持小鼠肝细胞功能的必要基因,Cre-LoxP技术可成功构建HuRLKO小鼠模型,且特异性高,表型明显,为研究HuR在肝脏各项疾病的发生发展中所扮演的角色,提供了一个很好的疾病模型。

关键词:

Abstract:

Objective To investigate the effect of hepatocyte-specific HuR gene knockout on liver function in mice model. Methods LoxP-labeled human antigen R (HuR) mice (HuRfl/fl) and Alb-Cre transgenic mice were imported from the Jackson Laboratory in the United States. After several generations of crossing and breeding,hepatocyte-specific HuR gene knockout mice (HuRLKO) model was established. PCR technology was used to identify the genotypes of mice. Western blotting and immunofluorescence experiments were used to examine the expression of HuR in mice liver. HE staining was used to observe the structural changes in mice liver. HuRfl/fl/Alb-Cre and HuRfl/fl mice were fed with high-fat diet, and the changes of body weight and liver weight were recorded. Serum was extracted to detect hepatic impairment and lipid metabolism related indexes. Results PCR detected the genotypes of the progeny mice successfully, including HuRfl/fl/Alb-Cre mice and HuRfl/fl mice. Western blotting and immunofluorescence experiments proved that the hepatocyte-specific HuR gene knockout in mice was successful. The results of HE staining suggested that the hepatocytes of HuRLKO mice had degeneration and local necrosis. The high-fat feeding experiment showed that the differences in liver weight, AST, ALT, HDL-C and blood glucose between the two groups of mice were statistically significant (P<0.05), but the difference in body weight, TG and LDL-C was not statistically significant (P>0.05). Conclusion HuR gene is an essential gene to maintain the function of hepatocytes in mice model. Cre-LoxP technology can successfully construct the HuRLKO mice model with high specificity and obvious phenotype, which provides a good disease model for the study of the role of HuR in the occurrence and development of various liver diseases.

Key words:

中图分类号: 

  • R657
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