畜牧兽医学报2025，Vol.56Issue(10)：4914-4924,11.DOI:10.11843/j.issn.0366-6964.2025.10.013
miR-376c-5p靶向PIK3CA对前体脂肪细胞脂肪沉积的影响
miR-376c-5p Regulates Lipid Accumulation in Preadipocytes by Targeting PIK3CA
摘要
Abstract
This study aimed to investigate the targeting relationship between miR-376c-5p and PIK3CA to elucidate how miR-376c-5p influences 3T3-L1 preadipocyte differentiation,exploring its role and regulatory mechanisms in adipogenesis.In the early stage,the research group selected 6 healthy 3-year-old Bashbay sheep(large-tailed type)and 6 F2 generation sheep of wild Argali × Bashbay sheep(small-tailed type)under the same feeding conditions,their tail adipose tissues were col-lected,and screened out the differentially expressed miR-376c-5p through the sequencing data analysis results of the tail fat.Then,an miR-376c-5p mimic overexpression vector was construc-ted for transfection,and the expression levels of PIK3CA and adipogenic differentiation marker genes were detected by qPCR.The adipogenic capacity was detected by combining Oil Red O staining and Bodipy staining.The conservation of miR-376c-5p in different species was analyzed,and bioinformatics software was used to predict the target gene(PIK3CA)of miR-376c-5p and the binding sites between them.Additionally,dual-luciferase reporter assay was performed to verify the targeting relationship between miR-376c-5p and PIK3CA.Compared with the mimic NC group,qPCR results showed that overexpression of miR-376c-5p significantly downregulated the mRNA expression levels of key adipogenic differentiation marker genes(PPARγ,ADIPOQ,ACACA and FASN)(P＜0.001).Both Oil Red O and Bodipy staining results demonstrated a reduction in lipid droplet formation following miR-376c-5p overexpression,indicating that over-expression of miR-376c-5p inhibited adipogenic differentiation in 3T3-L1 preadipocytes.Bioinfor-matic prediction revealed a potential binding site between miR-376c-5p and the 3'UTR of PIK3CA.Dual-luciferase reporter assay further confirmed that overexpression of miR-376c-5p significantly suppressed the luciferase activity of a vector containing the PIK 3CA 3'UTR frag-ment(P＜0.05).Moreover,overexpression of miR-376c-5p markedly reduced the mRNA ex-pression level of the candidate target gene PIK3CA(P＜0.05).These findings suggest that miR-376c-5p may inhibit adipogenesis by targeting PIK 3CA,providing a theoretical basis for further exploration of miRNA functions at the molecular level.关键词
miR-376c-5p/PIK3CA/脂肪沉积Key words
miR-376c-5p/PIK3CA/fat deposition分类
畜牧业引用本文复制引用
宋湘怡,曹行,刘武军,蒋琳..miR-376c-5p靶向PIK3CA对前体脂肪细胞脂肪沉积的影响[J].畜牧兽医学报,2025,56(10):4914-4924,11.基金项目
天山英才计划(2023TSYCLJ0017) （2023TSYCLJ0017）
自治区羊产业技术体系(XJARS-09-04) （XJARS-09-04）
