中国药理学通报2024，Vol.40Issue(1)：189-194,6.DOI:10.12360/CPB202310009
肝星状细胞特异性Grk2基因敲除小鼠模型的制备及鉴定
Establishment and genotype identification of hepatic stellate cell-specific Grk2 gene knockout mouse model
摘要
Abstract
Aim To establish a stable hepatic stellate cell(HSC)-specific G protein-coupled receptor kinase 2(GRK2)knockout mice and provide the important animal model for fur-ther studying the biological function of GRK2 in HSC.Methods The loxP-labeled Grk2 gene mouse(Grk2fl/fll)and the Lrat-Cre mouse were hybridized to establish a HSC-specific Grk2 gene knockout mouse(Grk2△HSC)model.The growth and repro-duction of mice were observed and analyzed.The flox and Cre gene were amplified by PCR for genotyping.Immunofluorescence double staining was used to examine the expression of GRK2 in HSC.Western blot was used to examine the expression of GRK2 in primary HSC,lung,spleen,kidney and heart tissue.HE staining was used to observe the structural changes in mice liver,lung,spleen,heart and kidney tissues.Results The genotypes of Grk2△HSC mice were successfully identified.No significant differences were detected in diet,drinking water,reproductive ability between the two groups.The results of immunofluores-cence double staining and Western blot showed that the expres-sion of GRK2 in HSC was significantly lower in the Grk2△HSC mice than that in the control mice,and the expression of GRK2 in other major tissue such as lung,spleen,kidney and heart of Grk2△HSC mice had no significant changes compared with the con-trol group.HE staining showed that there were no significant differences in structures of liver and other main tissue,which could be used for the follow-up study.Conclusions This study successfully establishes HSC specific Grk2 gene knockout mice using Cre-loxP technology,providing an excellent tool for further studying the role of GRK2 in liver.关键词
G蛋白偶联受体激酶2/Cre-loxP重组酶系统/细胞特异性敲除/肝星状细胞/基因鉴定/繁育Key words
G protein-coupled receptor kinase 2/Cre-loxP recombinase system/cell specific knockout/hepatic stellate cell/genotyping identification/breeding分类
医药卫生引用本文复制引用
王语涵,许雅萍,李南,陈婷婷,李玲,高萍萍,王华,魏伟,孙妩弋..肝星状细胞特异性Grk2基因敲除小鼠模型的制备及鉴定[J].中国药理学通报,2024,40(1):189-194,6.基金项目
国家自然科学基金资助项目(No 82370632) （No 82370632）
安徽省高校杰出青年科研项目(No 2023AH020033) （No 2023AH020033）
安徽省转化医学研究院科研基金项目(No 2022zhyx-C07) （No 2022zhyx-C07）
安徽医科大学科研水平提升计划(No 2021xkjT016) （No 2021xkjT016）
安徽医科大学第三附属医院基础与临床合作研究提升计划培育专项(No 2022sfy014) （No 2022sfy014）
