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中华肝脏外科手术学电子杂志 ›› 2019, Vol. 08 ›› Issue (03) : 270 -275. doi: 10.3877/cma.j.issn.2095-3232.2019.03.021

所属专题: 文献

基础研究

Cre-LoxP技术构建肝脏特异性HIF-2α基因敲除小鼠模型
赵安竹1, 付辉蓉1, 李昀1, 陈坚娣1, 鲁红云1,()   
  1. 1. 519000 珠海,中山大学附属第五医院老年病科
  • 收稿日期:2019-02-26 出版日期:2019-06-10
  • 通信作者: 鲁红云
  • 基金资助:
    国家自然科学基金(81670815)

Establishment of a mouse model with liver-specific HIF-2α gene knockout with Cre-loxP technique

Anzhu Zhao1, Huirong Fu1, Yun Li1, Jiandi Chen1, Hongyun Lu1,()   

  1. 1. Department of Gerontology, the Fifth Affiliated Hospital of Sun Yat-sen University, Zhuhai 519000, China
  • Received:2019-02-26 Published:2019-06-10
  • Corresponding author: Hongyun Lu
  • About author:
    Corresponding author: Lu Hongyun, Email:
引用本文:

赵安竹, 付辉蓉, 李昀, 陈坚娣, 鲁红云. Cre-LoxP技术构建肝脏特异性HIF-2α基因敲除小鼠模型[J]. 中华肝脏外科手术学电子杂志, 2019, 08(03): 270-275.

Anzhu Zhao, Huirong Fu, Yun Li, Jiandi Chen, Hongyun Lu. Establishment of a mouse model with liver-specific HIF-2α gene knockout with Cre-loxP technique[J]. Chinese Journal of Hepatic Surgery(Electronic Edition), 2019, 08(03): 270-275.

目的

探讨采用Cre-LoxP技术构建肝脏特异性HIF-2α基因敲除小鼠模型的可行性。

方法

由美国Jackson实验室引进loxP标记的HIF-2α基因小鼠,采用Cre/loxP重组酶系统和Alb-Cre转基因小鼠,通过多次繁殖杂交,建立肝脏特异性HIF-2α基因敲除小鼠模型。利用PCR技术鉴别小鼠基因型,并用RT-qPCR检测小鼠肝脏、脂肪及肌肉组织HIF-2α mRNA水平,同时检测野生型小鼠和Alb-Cre+/HIF-2αfl/fl的纯合子小鼠血糖、血脂以及肝功能水平。3组HIF-2α mRNA数据比较采用单因素方差分析和Turkey检验。

结果

PCR法成功检测出子代小鼠基因型,包括Alb-Cre+/HIF-2αfl/fl和Alb-Cre-/HIF-2αfl/fl纯合子小鼠。肝脏HIF-2α基因敲除鼠肝脏组织HIF-2α mRNA表达量为0.10±0.02,明显低于野生型小鼠的1.00±0.10(HSD=-1.87,P<0.05)。

结论

采用Cre-LoxP技术可成功构建肝脏特异性HIF-2α基因敲除小鼠。

Objective

To investigate the feasibility of establishing a mouse model with liver-specific HIF-2α gene knockout using Cre-LoxP technique.

Methods

The loxP-labeled mice with HIF-2α gene were introduced from the Jackson Laboratory of the United States. A liver-specific HIF-2α gene knockout mouse model was established by using Cre/loxP recombinase system and Alb-Cre transgenic mice through multiple generations of hybridization. The genotypes of mouse were identified by PCR. The expression levels of HIF-2α mRNA in the liver, fat and muscular tissues were detected by RT-qPCR. The blood glucose, blood lipid and liver function levels in the wild-type mice and Alb-Cre+/HIF-2αfl/fl homozygous mice were detected. The levels of HIF-2α mRNA among three groups were statistically compared by one-way ANOVA and Turkey test.

Results

The genotypes of the 2nd generation mouse were successfully identified, including Alb-Cre+/HIF-2αfl/fl and Alb-Cre-/HIF-2αfl/fl homozygous mice. The expression level of HIF-2α mRNA in the liver tissues of HIF-2α gene knockout mice was 0.10±0.02, significantly lower than 1.00±0.10 of the wild-type mice (HSD=-1.87, P<0.05).

Conclusion

The mouse models with liver-specific HIF-2α gene knockout can be successfully established using Cre-LoxP technique.

表1 基因鉴定引物
表2 RT-qPCR引物
图1 HIF-2α基因敲出小鼠基因鉴定电泳图
图2 不同基因型小鼠肝脏、脂肪和肌肉组织HIF-2α基因转录水平
图3 不同基因型小鼠IPGTT和IPITT结果
图4 不同基因型小鼠血脂水平
图5 不同基因型小鼠肝功能水平
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