中国实验动物学报2024，Vol.32Issue(4)：493-502,10.DOI:10.3969/j.issn.1005-4847.2024.04.010
肝特异性Rbp4基因敲除小鼠的建立及糖代谢特征分析
Establishment of liver-specific Rbp4 knockout mice and analysis of glucose metabolism characteristics
摘要
Abstract
Objective To establish a liver-specific Rbp4 gene knockout mouse model and to explore the effect of liver Rbp4 gene deletion on glucose metabolism.Methods Cre-LoxP technology was used to construct a liver-specific Rbp4 gene knockout mouse model using C57/BL6J and Alb-Cre mice.The genotype of the mice was identified by polymerase chain reaction and agarose gel electrophoresis.Ten 18 week old C57/BL6J male mice were included in the WT group,10 flox homozygous and Alb-Cre negative mice of the same age were included in the experimental control group(Rbp4flox/flox:Cre-),and 10 flox homozygous and Alb-Cre positive mice of the same age were included in the experimental group(Rbp4flox/flox:Cre+).Expression levels of RBP4 protein and mRNA in the liver were verified by Western Blot and quantitative reverse transcription-polymerase chain reaction(qRT-PCR),respectively,and expression levels of Rbp4 mRNA in other tissues were detected by qRT-PCR.Morphological changes in liver tissue were detected by hematoxylin and eosin staining.Blood glucose values were detected in mouse tail vein blood samples using a blood glucose meter,and glucose tolerance and insulin tolerance were determined.Expression levels of the liver glucose metabolism genes phosphoenolpyruvate carboxylase(Pepck)and glucose-6-phosphatase(G6pase)were detected by qRT-PCR.Results Liver-specific Rbp4 knockout mice were successfully bred and identified.RBP4 protein and mRNA levels were significantly decreased in the liver of Rbp4flox/flox:Cre+mice(P＜0.05),but there was no significant difference in the relative expression levels of Rbp4 mRNA in fat,kidney,pancreas,spleen,heart,or muscle tissues among the three groups(P＞0.05).Liver-specific Rbp4 knockout had no significant effect on liver morphology,glucose tolerance,or insulin tolerance(P＞0.05).Pepck mRNA levels in the liver differed significantly among the three groups(P＜0.05),and pairwise comparison showed that liver Pepck mRNA levels were significantly lower in Rbp4flox/flox:Cre+mice compared with levels in Rbp4flox/flox:Cre-mice(P＜0.05).There was no significant difference in liver glucose-6-phosphatase(G6pase)mRNA expression among the three groups(P＞0.05).Conclusions We successfully constructed a liver-specific Rbp4 knockout mouse model.Deletion of Rbp4 in the liver inhibited expression of Pepck mRNA in the liver,thus providing a basis for further exploration of the role of this gene in glucose metabolism in mice.关键词
2型糖尿病/视黄醇结合蛋白4/基因敲除/糖代谢/胰岛素抵抗Key words
type 2 diabetes mellitus/retinol-binding protein 4/gene knockout/glucose metabolism/insulin resistance分类
生物科学引用本文复制引用
卢婉贤,马琦,王黎,刘梦迪,郭宝平..肝特异性Rbp4基因敲除小鼠的建立及糖代谢特征分析[J].中国实验动物学报,2024,32(4):493-502,10.基金项目
国家自然科学基金(82060589). Funded by the National Natural Science Foundation of China(82060589). （82060589）
