Construction and application of inducible macrophage-specific knockout GRK2 gene mice model
10.19405/j.cnki.issn1000-1492.2023.04.003
- Author:
Qi Wei
1
;
Xuemin Zhu
1
;
Xiaoyi Liu
1
;
Xuezhi Yang
1
;
Wei Wei
1
Author Information
1. Institute of Clinical Pharmacology，Anhui Medical University，Key Laboratory of Anti-inflammatory Immune Medicine，Ministry of Education，Anhui Collaborative Innovation Center for Anti-inflammatory and Immuno Drugs，Hefei 230032
- Publication Type:Journal Article
- Keywords:
G protein-coupled receptor kinase 2;
Cre / LoxP system;
specific knockout mice;
macrophages;
identifi- cation of genotype
- From:
Acta Universitatis Medicinalis Anhui
2023;58(4):534-540
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To establish an inducible macrophage-specific knockout G protein-coupled receptor kinase 2 ( GRK2) gene ( GRK2flox / flox Lyz2-CreERT + ) mice model．
Methods:GRK2flox / flox Lyz2-CreERT + mice were constructed based on Cre / LoxP system．The genotypes of GRK2flox / floxLyz2-CreERT + mice were identified by PCR amplification and agarose gel electrophoresis．After the mice were sacrificed by carbon dioxide method，the expression of GRK2 in bone marrow-derived macrophages ( BMDMs) and peritoneal macrophages ( PMs) was detected by Western blot．Immunofluorescence was used to detect GRK2 expression in mouse brain，heart and spleen macrophages．The M1 / M2 ratio in PMs induced by prostaglandin E2 (PGE2) was analyzed by flow cytometry.
Results:The results of genotype identification showed that the mice with a band at 355 bp in the length of the flox amplification product and a band at 355 bp in the length of the Cre amplification product were GRK2flox / floxLyz2-CreERT + mice．Western blot results showed that GRK2 expression in BMDMs and PMs of GRK2flox / floxLyz2-CreERT + mice decreased compared with GRK2flox / flox mice(P＜0. 01) ．Immunofluorescence results showed that GRK2 expression decreased in the brain，heart and spleen of GRK2flox / floxLyz2-CreERT + mice compared with GRK2flox / flox mice (P < 0. 01) ．Flow cytometry showed that compared with GRK2flox / flox mice，there was no significant difference in the proportion of CD86 / CD206 in the PMs of GRK2flox / floxLyz2-CreERT + mice．Under PGE2 ( 10 μmol / L) stimulation， the proportion of CD86 / CD206 in GRK2flox / floxLyz2-CreERT + mice PMs increased (P ＜0. 01) ．The proportion of CD86 / CD206 in the PMs of GRK2flox / flox mice was higher than that of GRK2flox / floxLyz2-CreERT + mice(P＜0. 01) .
Conclusion :In this study，GRK2flox / floxLyz2-CreERT + mice model was successfully constructed，and the mice promoted PGE2-induced polarization of PMs to M2-type macrophages compared with control mice.
- Full text:2024071710011124343诱导型巨噬细胞特异性敲除G...2基因小鼠模型的构建及应用_魏琦.pdf
