肝星状细胞特异性Grk2基因敲除小鼠模型的制备及鉴定OACSTPCD

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.