动物造模,巨噬细胞 z-bio 2021-06-29 977

　　Objective: To construct and identify Atg5 gene knockout mice under the condition of macrophages, and to provide an animal model for studying the role of macrophage autophagy in the pathogenesis of kidney disease.

　　Method: The introduced LysM-Cre mice were bred with Atg5flox/+ and selfed Atg5flox/+ mice to obtain the progeny of Atg5flox/+Cre+/- and Atg5flox/floxCre-/-, respectively. Mice; Mating offspring mice. Compare the above two genotypes to obtain macrophage knockout mice (Atg5flox/floxCre +/-, Atg5-/-) and control mice (Atg5flox/floxCre-/-), Atg5 +/+). Extract genomic DNA from mouse tail tissues, PCR amplification and agarose gel electrophoresis, and perform genotyping of mice at the DNA level. The macrophage NA and protein were extracted from mouse bone marrow, and the knockout effect of Atg5 gene was tested using gene sequencing and western blotting.

　　Result: The Atg5 gene knockout mouse model under macrophage conditions was successfully established, and the mice survived and were fertile. At the gene and protein level, the macrophage Atg5 gene was successfully knocked out, and the basal level of autophagy in Atg5 mice was lower than that in Atg5 mice (p62 was significantly increased, and LC3II was significantly decreased). Normal basic level.

　　Conclusion: This study used the Cre/loxp system to successfully construct and identify conditional macrophage Atg5 gene knockout mice, to study the role of macrophage autophagy in the pathogenesis of kidney disease from the animal level, providing a study for the study platform.