农业生物技术学报2025，Vol.33Issue(6)：1312-1323,12.DOI:10.3969/j.issn.1674-7968.2025.06.012
山羊PDE4B表达特性分析及其对肌内前体脂肪细胞分化的影响
Expression Characteristics Analysis of Goat(Capra hircus)Phosphodiesterase 4B(PDE4B)and Its Effect on the Differentiation of Intramuscular Precursor Adipocytes
摘要
Abstract
Intramuscular fat(IMF)in goat(Capra hircus)is an important factor in affecting meat quality traits such as flavor and juiciness.This study focused on the cloning and functional analysis of phosphodiesterases 4B(PDE4B),to delineate its regulatory role in modulating the differentiation dynamics of caprine intramuscular preadipocytes.In this study,the intramuscular precursor adipocytes of JianZhou big-eared goats were used to amplify the target gene fragments by RT-PCR,and their sequences were analyzed by online bioinformatics tools.The expression level of PDE4B gene in different tissues and different differentiation stages of intramuscular precursor adipocytes in goats was detected using qPCR.The PDE4B overexpression plasmid(pEGFP-N1-PDE4B)was successfully engineered via molecular subcloning and subsequently transfected into caprine intramuscular preadipocytes.Subsequent Oil red O staining and qPCR assays were employed to assess its regulatory effects on intracellular lipid droplet deposition,coupled with transcriptional profiling of key adipogenic differentiation,triglyceride biosynthesis,and lipolytic metabolism markers.The results showed that the length of the cloned PDE4B gene was 1815 bp,including the coding sequence(CDS)region of 1 629 bp,which encoded a total of 542 amino acids.Furthermore,the expression of PDE4B gene was significantly higher in goat lung than in other tissues examined(P＜0.01),and reached the highest expression after 48 h induction of differentiation in intramuscular adipocytes(P＜0.01).Oil red O staining and the quantitative results after PDE4B overexpression showed that there were significant increase in lipid droplet aggregation(P＜0.05).The mRNA expression level of differentiation marker genes CCAAT/enhancerbindingprotein α(C/EBPα),C/EBPβ and triglyceride synthesis marker genes activator protein 2(AP2)and fatty acid synthase(FASN)were extremely significantly up-regulated after overexpression of PDE4B gene(P＜0.01).Meanwhile,the expression of triglyceride catabolism marker genes adipose triglyceride lipase(ATGL),hormone-sensitive lipase(HSL)and lipoprotein lipase(LPL)were significantly down-regulated(P＜0.01).In conclusion,PDE4B gene might positively regulate the process of intramuscular precursor adipocyte differentiation in goats,and this effect might be realized by up-regulating the expression of C/EBPα,C/EBPβ,AP2,and FASN.This study provides data support for revealing the molecular mechanism of intramuscular precursor adipocyte differentiation regulated by PDE4B in goats.关键词
山羊/磷酸二酯酶4B(PDE4B)/表达特性/肌内前体脂肪细胞/脂质沉积调控Key words
Goat/Phosphodiesterases 4B(PDE4B)/Expression characteristics/Intramuscular precursor adipocyte/Lipid deposition regulation分类
农业科技引用本文复制引用
李金岚,孙诗雨,何晓菲,张力懿,林亚秋,刘伟,邢佳妮..山羊PDE4B表达特性分析及其对肌内前体脂肪细胞分化的影响[J].农业生物技术学报,2025,33(6):1312-1323,12.基金项目
国家自然科学基金面上项目(32372857) （32372857）
四川省青年科技创新研究团队(2022JDTD0030) （2022JDTD0030）
中央高校基本科研业务费专项基金项目(ZYN2024015) （ZYN2024015）
