局解手术学杂志2025，Vol.34Issue(11)：950-954,5.DOI:10.11659/jjssx.01E025013
肝细胞特异性NLRP3基因敲除小鼠模型的构建及鉴定
Construction and identification of hepatocyte-specific NLRP3 gene knockout mouse model
摘要
Abstract
Objective To explore the possibility and genetic identification method of constructing a hepatocyte-specific NLRP3 gene knockout mouse model by using Cre-LoxP system gene knockout technology.Methods Phase one:mice specifically expressing the albumin promoter-Cre(AlbCre)recombinase in hepatocytes were mated with NLRP3flox/flox mice,and the hepatocyte-specific NLRP3 gene knockout mice with the genotype of NLRP3flox/flox/AlbCre+/-(hepatocyte NLRP3 knockout group)and the control mice in the same litter with the genotype of NLRP3flox/flox/AlbCre-/-(control group in the same litter)were obtained after two generations of selection and mating.The second stage was the mass reproduction stage.Mating NLRP3flox/flox/AlbCre+/-target mice with NLRP3flox/flox mice could quickly obtain a large number of experimental target mice and control mice in the same litter.The DNA was extracted from the tails of mice after numbering,and the offspring genotype was identified by PCR.qPCR and Western blot were used to detect the mRNA and protein expression levels of NLRP3 gene in the liver tissue.HE staining was used to observe the morphological changes in liver tissues,and serum liver transaminases and inflammatory factors were detected.The changes in body weight,liver-to-body ratio and special circumstances during reproduction and development of mice in the two groups were observed.Results The offspring genotype of the target mice in the F2 generation was consistent with theoretical result of NLRP3flox/flox/AlbCre+/-.The mRNA and protein levels of NLRP3 in liver tissues of mice in the hepatocyte NLRP3 knockout group were significantly lower than those in the control group in the same litter(P<0.05).The mice in the hepatocyte NLRP3 knockout group was not affected in terms of growth,development and reproduction after the NLRP3 gene knockout.There were no statistically significant differences in the body weight,liver-to-body ratio,liver tissue morphology,serum liver transaminase or inflammatory factors between the hepatocyte NLRP3 knockout group and the control group in the same litter(P>0.05).Conclusion The Cre-LoxP gene knockout technology can be used to successfully construct a hepatocyte-specific NLRP3 gene knockout mouse model,providing an important technical support for the next step of studying the function of the NLRP3 gene in the liver at the animal level.关键词
NLRP3/Cre-LoxP技术/肝/基因敲除/肝细胞Key words
NLRP3/Cre-LoxP technology/liver/gene knockout/hepatocyte分类
医药卫生引用本文复制引用
苟鸿翔,韩金成,甘丰德,易耀兴,范克瑞,胡凯..肝细胞特异性NLRP3基因敲除小鼠模型的构建及鉴定[J].局解手术学杂志,2025,34(11):950-954,5.基金项目
2023年重庆医科大学大学生科学研究与创新实验项目(202323) （202323）
重庆市博士"直通车"科研项目基础研究与前沿探索(CSTB2022BSXM-JCX0043) （CSTB2022BSXM-JCX0043）
