中国农业大学学报2024，Vol.29Issue(4)：226-238,13.DOI:10.11841/j.issn.1007-4333.2024.04.20
FOXO1基因调控牛前体脂肪细胞增殖和分化的功能研究
Study on the function of FOXO1 gene in regulating the proliferation and differentiation of bovine preadipocytes
摘要
Abstract
The aim of the study was to investigate the effects of interfering with Forkhead box protein O1(FOXO1)gene on the proliferation and differentiation of bovine preadipocytes.Taking the Guyuan cattle preadipocytes as the research object,bovine preadipocytes were isolated by tissue block method,and the purity and differentiation potential of the isolated cells were identified by phenotypic identification and marker gene expression profiling.The bovine FOXO1 interfering lentivirus was screened and packaged,and the effects of interfering FOXO1 on the proliferation and differentiation of bovine adipocytes were explored by EdU staining,flow cytometry,oil red O staining and qPCR.The results showed that:1)The bovine preadipocytes were successfully isolated and showed high purity and differentiation potential.2)EdU staining showed that infection of Lv-FOXO1 significantly increased the proliferation rate of bovine adipocytes(P＜0.01).3)Flow cytometry results showed that interfering with FOXO1 significantly increased the proportion of S-phase positive cells and promoted the G1/S phase transformation of bovine adipocytes,which in turn promoted the proliferation of bovine adipocytes.4)After interfering with FOXO1,the expression levels of proliferation marker genes PCNA,CDK1 and CDK2 were all extremely significantly upregulated(P＜0.01).5)After interfering with FOXO1,the production of lipid droplets in bovine precursor adipocytes was significantly reduced,and the expression of PPARG,a lipid-forming marker gene,was extremely significantly down-regulated(P＜0.01),together with the expressions of CEBPB and LPL(P＜0.05).In summary,interfering with FOXO1 can promote the G1/S phase transformation of bovine adipocytes by up-regulating the expressions of proliferation marker genes PCNA,CDK1 and CDK2,thereby promoting the proliferation of bovine adipocytes.Moreover,it can reduce the formation of lipid droplets in bovine adipocytes by reducing the expressions of lipogenic marker genes PPARG,CEBPB and LPL,thus affecting fat deposition.This study can provide theoretical basis for genetic improvement of meat quality of Chinese local yellow cattle.关键词
FOXO1/固原黄牛,前体脂肪细胞/增殖/分化Key words
FOXO1/Guyuan cattle,preadipocytes/proliferation/differentiation分类
农业科技引用本文复制引用
宋雅萍,张久盘,魏大为,杨东梅,赵毅昂,姜超,宋小雨,吴昊..FOXO1基因调控牛前体脂肪细胞增殖和分化的功能研究[J].中国农业大学学报,2024,29(4):226-238,13.基金项目
国家自然科学基金项目(32060744,32202641) （32060744,32202641）
宁夏自然科学基金项目(2023AAC03394) （2023AAC03394）
宁夏重点研发计划项目(2023BCF01006) （2023BCF01006）
