Gpr183基因敲除小鼠的构建及其在急性肝损伤中功能初步分析

吴学文; 尹思琪; 钱颖; 杜冰; 秦居亮; 陈金联

doi:10.16016/j.1000-5404.202103181

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Gpr183基因敲除小鼠的构建及其在急性肝损伤中功能初步分析

基础医学 | 更新时间：2026-01-09

- Gpr183基因敲除小鼠的构建及其在急性肝损伤中功能初步分析
- Construction of GPR183 knockout mice and its function in acute liver injury
- 第三军医大学学报 2021年43卷第18期页码：1769-1774
- 作者机构：
  
  1.232001 安徽淮南，安徽理工大学医学院
  2.201100 上海，华东师范大学生命科学学院生命医学研究所
- 作者简介：
  
  [陈金联，E-mail：xuewenmed@163.com CHEN Jinlian, E-mail: xuewenmed@163.com
- 基金信息：
- DOI：10.16016/j.1000-5404.202103181
  中图分类号： R-332;R394-33;R575
- 收稿：2021-03-22，
  
  修回：2021-06-01，
  
  纸质出版：2021-09-30
- 稿件说明：
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吴学文, 尹思琪, 钱颖, 等. Gpr183基因敲除小鼠的构建及其在急性肝损伤中功能初步分析[J]. 第三军医大学学报, 2021,43(18):1769-1774.

Xuewen WU, Siqi YIN, Ying QIAN, et al. Construction of GPR183 knockout mice and its function in acute liver injury[J]. Journal of Third Military Medical University, 2021, 43(18): 1769-1774.
吴学文, 尹思琪, 钱颖, 等. Gpr183基因敲除小鼠的构建及其在急性肝损伤中功能初步分析[J]. 第三军医大学学报, 2021,43(18):1769-1774. DOI： 10.16016/j.1000-5404.202103181.

Xuewen WU, Siqi YIN, Ying QIAN, et al. Construction of GPR183 knockout mice and its function in acute liver injury[J]. Journal of Third Military Medical University, 2021, 43(18): 1769-1774. DOI： 10.16016/j.1000-5404.202103181.

摘要

目的

利用成簇的规律间隔短回文重复序列和相关蛋白9(CRISPR/Cas9)基因编辑技术构建Gpr183全基因敲除小鼠模型，并对Gpr183受体在淋巴细胞的调控功能和在急性肝损伤过程中的作用进行了初步分析。

方法

根据CRISPR/Cas9基因编辑技术实验方法构建Gpr183全基因敲除纯合子F2代小鼠，随后通过Western blot方法检测野生型和敲除型小鼠肝脏和脾脏组织Gpr183基因蛋白表达水平，并通过免疫荧光(IF)技术分析了野生型和敲除型小鼠脾脏淋巴细胞的分布情况以及IL-17和IL-22的表达水平，最后我们对Gpr183受体敲除后在小鼠急性肝损伤模型中进行了初步分析？。

结果

Gpr183基因敲除小鼠可正常生长繁殖，F2代纯合子小鼠(KO)较野生小鼠(WT)相比基因片段部分缺失，同时Western blot结果显示KO小鼠肝脏、脾脏组织中Gpr18

3基因表达蛋白缺失，IF结果显示Gpr183受体可引起小鼠脾脏B淋巴细胞由滤泡中央区向滤泡边缘区迁移，并且减弱了脾脏淋巴细胞IL-17和IL-22的分泌功能(

0.05)，同时在小鼠急性肝损伤诱导模型中，我们发现Gpr183受体敲除小鼠能明显缓解小鼠急性肝损伤过程(

0.01)，差异具有统计学意义。

结论

成功构建了Gpr183受体敲除小鼠模型，为Gpr183基因功能在体内研究提供了稳定的小鼠敲除模型，同时Gpr183受体敲除后能减轻小鼠急性肝损伤。

Abstract

Objective

To construct a GPR183 knockout mouse model by genome editing using clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) system

and to analyze the role of GPR183 receptor in lymphocyte regulation and acute liver injury.

Methods

Firstly

GPR183 full-gene knockout homozygous F2 generation mice were constructed using CRISPR/Cas9 gene editing technique. Then

the expression level of GPR183 protein in the liver and spleen tissues of wild-type (WT) and knockout (KO) mice was detected by Western blotting and the Results were compared; the distribution of spleen lymphocytes and the expression of IL-17 and IL-22 in WT and KO mice were analyzed by immunofluorescence (IF) assay. Finally

the function of GPR183 knockout was explored in mouse model with acute liver injury.

Results

GPR183 KO mice could grow and reproduce normally

and F2 homozygous KO mice had partial deletion of gene fragments as compared to WT mice. Meanwhile

Western blotting showed that GPR183 expression was missing in the liver and spleen tissues of KO mice. In addition

the Results of IF assay suggested that GPR183 receptor induced the migration of spleen B lymphocytes from the central to the peripheral region of the follicles

and reduced the secretion of IL-17 and IL-22 by the spleen lymphocytes (

0.05). We also found that GPR183 KO mice significantly alleviated the process of acute liver injury in mice (

0.01).

Conclusion

A GPR183 knockout mouse m

odel is successfully established

which provides a stable animal model for the further study of GPR183 gene function

in vivo

. Moreover

the knockout of GPR183 receptor can alleviate acute liver injury in mice.

关键词

Keywords

references

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