安徽医科大学学报2025，Vol.60Issue(8)：1432-1439,8.DOI:10.19405/j.cnki.issn1000-1492.2025.08.010
条件性胰岛β细胞HLF基因敲除小鼠构建与鉴定
Construction and identification of conditional HLF knockout mice with islet β cells
摘要
Abstract
Objective To explore the mechanism of action of hepatic leukemia factor(HLF)in diabetes mellitus and to construct a conditional animal model of mice with islet β-cell-specific HLF gene knockout.Methods At the cellular level,the effects of HLF inhibition or overexpression on the proliferation of MIN6 cells was verified by the CCK-8 assay.The effects of HLF inhibition or overexpression were detected at the mRNA level and protein level by RT-qPCR technology and Western blot technology,respectively.HLFflox/flox transgenic mice were crossed with Pdx1-Cre+/-mice(C57BL/6J)to obtain offspring mice.The genotypes of the mice were identified by the PCR method.The differences in the expression levels of the HLF gene at the mRNA and protein levels in islet β-cell knockout mice(HLFflox/flox Cre+/-)and control mice(HLFflox/flox)were detected by RT-qPCR technology and Western blot technology to verify the knockout effect.At the same time,the islet tissues of the mice in two groups were taken to make paraffin sections and analyzed by hematoxylin-eosin(HE)staining.Results HLF gene inhibition or overex-pression had no significant effect on the proliferation of MIN6 cells.When the HLF gene was inhibited in MIN6 cells,the mRNA expression level decreased by 74％compared with the control group,and the protein expression level decreased by 60％compared with the control group.After overexpressing the HLF gene,the mRNA expres-sion level was 2.13 times compared with that of the control group,and the protein expression level was 1.8 times compared with that of the control group.The mRNA expression level of the HLF gene in the knockout mice de-creased by 89％compared with the control group,and the protein expression level decreased by 65％compared with the control group.The results of HE staining showed that there was no significant difference in the cell mor-phology in the islet tissues between the knockout mice and the control mice.Inhibiting HLF increased the glycogen content in MIN6 cells by approximately 20％.Conclusion The HLF gene knockout mice are successfully con-structed,providing an animal model for studying the role of HLF in the pathogenesis of diabetes mellitus.关键词
肝白血病因子/条件性胰岛β细胞/RT-qPCR/Western blot/基因敲除/C57BL/6J小鼠Key words
hepatic leukemia factor/conditional islet β cells/RT-qPCR/Western blot/gene knockout/C57BL/6J mice分类
医药卫生引用本文复制引用
侯梦龙,齐心语,吴建凤,廖启超,马杰,周磊,李一星..条件性胰岛β细胞HLF基因敲除小鼠构建与鉴定[J].安徽医科大学学报,2025,60(8):1432-1439,8.基金项目
国家重点研发计划项目(编号:2023YFE0100800) National Key Research and Development Program of China(No.2023YFE0100800) （编号:2023YFE0100800）
