重庆医学2025，Vol.54Issue(3)：573-579,7.DOI:10.3969/j.issn.1671-8348.2025.03.003
miR-29b-3p介导HNF-4α抑制凝血因子X表达在创伤性凝血病的机制研究
Study on the mechanism of HNF-4α mediated by miR-29b-3p to inhibit the expression of coagulation factor Ⅹ in trauma induced coagulopathy
摘要
Abstract
Objective To investigate the function and mechanism of miR-29 family in trauma induced coagulopathy(TIC).Methods Bioinformatics was used to analyze the targeting relationship between miR-29 family members and hepatocyte nuclear factor-4α(HNF-4α).HE staining results,TEG parameters and coagu-lation parameters were used to verify the TIC rat model construction.Real-time quantitative fluorescent PCR(RT-qPCR)and Western blot were used to detect the expression of miR-29 family,HNF-4α and coagulation factor Ⅹ(FⅩ)in rat liver tissues.Overexpression of miR-29b-3p(miR-29b-3p mimics)or silence of miR-29b-3p(miR-29b-3p inhibitor)was transfected into hepatocytes,and the levels of miR-29b-3p,HNF-4α and FⅩ in hepatocytes were detected by RT-qPCR and Western blot.Double lucifase reporter gene assay verified the targeted regulation of miR-29b-3p on HNF-4α.The miR-29b-3p mimics and/or HNF-4α overexpression vector were transfected into hepatocytes,and the levels of miR-29b-3p,HNF-4α and FⅩ in hepatocytes were detected by RT-qPCR and Western blot.Results Bioinformatic prediction results from the miRDIP database identified that multiple members of the miR-29 family(miR-29a-3p,miR-29b-3p,miR-29b-5p,and miR-29c-3p)contain potential binding sites with HNF-4α.Histopathological evaluation through HE staining,combined with TEG parameters and coagulation profiles,confirmed successful establishment of the TIC rat model.Quantitative analyses using RT-qPCR and Western blot revealed that compared to controls,both HNF-4α and coagulation FⅩ expression levels were markedly suppressed in the model group,while miR-29b-3p expression showed significant elevation in TIC rats(P＜0.01).In vitro functional studies demonstrated that neither overexpression nor silencing of miR-29b-3p significantly influenced hepatocyte proliferation(P＞0.05).How-ever,forced expression of miR-29b-3p effectively downregulated HNF-4α and its downstream target FⅩ,whereas miR-29b-3p knockdown substantially upregulated these molecules(P＜0.05).This regulatory rela-tionship was further validated by dual luciferase reporter assays confirming direct targeting between HNF-4α and miR-29b-3p.Notably,exogenous HNF-4α overexpression significantly rescued FⅩ suppression induced by miR-29b-3p overexpression(P＜0.05).Conclusion miR-29b-3p is up-regulated in TIC,which can promote the progression of TIC by targeting HNF-4α to regulate FⅩ expression.关键词
创伤性凝血病/微小RNA/肝细胞核因子4α/凝血因子Ⅹ/生物信息学Key words
traumatic coagulopathy/miRNA/HNF-4α/FⅩ/bioinformatics分类
医药卫生引用本文复制引用
陈亮,贺慈,罗金花,黄志桃..miR-29b-3p介导HNF-4α抑制凝血因子X表达在创伤性凝血病的机制研究[J].重庆医学,2025,54(3):573-579,7.基金项目
湖南省卫生健康委科研计划项目(202217015220). （202217015220）
