bta-miR-19b靶向BCL3的验证及其在牛初乳免疫调控中的功能分析

doi:10.12160/j.issn.1672-5190.2026.03.003

畜牧与饲料科学 ›› 2026, Vol. 47 ›› Issue (3): 23-29.doi: 10.12160/j.issn.1672-5190.2026.03.003

bta-miR-19b靶向BCL3的验证及其在牛初乳免疫调控中的功能分析

孔繁霞¹, 王高峰², 赵濛³, 吴美灵¹, 王乐乐¹

1.乌兰察布医学高等专科学校,内蒙古集宁区 012000;
2.乌兰察布市动物疫病预防控制中心,内蒙古集宁区 012000;
3.内蒙古自治区农牧业科学院,内蒙古呼和浩特 010031

收稿日期:2026-03-27 出版日期:2026-05-30 发布日期:2026-08-27
通讯作者: 王乐乐（1985—）,女,副教授,博士,主要从事生物学教学和研究工作。
作者简介:孔繁霞（1982—）,女,讲师,主要从事生物学教学和研究工作。
基金资助:
内蒙古自然科学基金项目（2025LHMS03065）

Validation of bta-miR-19b Targeting BCL3 and Its Functional Analysis in Bovine Colostrum Immune Regulation

KONG Fanxia¹, WANG Gaofeng², ZHAO Meng³, WU Meiling¹, WANG Lele¹

1. Ulanqab Medical College, Jining District 012000, China;
2. Ulanqab Center for Animal Disease Prevention and Control, Jining District 012000, China;
3. Inner Mongolia Academy of Agricultural and Animal Husbandry Sciences, Hohhot 010031, China

Received:2026-03-27 Online:2026-05-30 Published:2026-08-27

摘要/Abstract

摘要： [目的] 旨在验证bta-miR-19b对B细胞白血病/淋巴瘤3（B-cell CLL/lymphoma 3,BCL3）基因的靶向关系,并解析bta-miR-19b介导牛初乳免疫调控的分子功能与作用机制。[方法] 采用TargetScan 和miRanda两个软件取交集筛选bta-miR-19b免疫相关靶基因,并通过RNAhybrid网站预测bta-miR-19b与免疫相关候选基因IL7R、EGFR、TNF、BCL3、NME7、MAPK8和MAP3K1的结合自由能（MFE）。以BCL3基因3′UTR预测结合位点为模板合成野生型（WT）、突变型（MT）片段,构建pmirGLO 荧光素酶报告载体;合成bta-miR-19b mimics（化学合成的双链RNA分子,模拟内源性miRNA功能）及NC对照（NC-mimics）,与报告载体共转染293T细胞用于靶向验证。并通过生物信息学方法筛选bta-miR-19b-BCL3轴在牛初乳免疫调控过程中生物学过程和关键通路。[结果] bta-miR-19b与 BCL3 的结合自由能最小,提示BCL3存在潜在结合可能。双荧光素酶报告基因实验结果证实bta-miR-19b能够特异性靶向结合BCL3基因的3′UTR 区。bta-miR-19b-BCL3调节轴共存在42个GO Term中,其中和免疫相关的生物学过程有24个。bta-miR-19b-BCL3调节轴参与的KEGG通路有肿瘤坏死因子TNF信号通路和C型凝集素受体信号通路。[结论] bta-miR-19b可靶向结合BCL3 3′UTR,鉴于BCL3是NF-kB关键调节因子,推测bta-miR-19b-BCL3-NF-κB-TNF 调节轴可能是牛初乳发挥免疫功能的核心分子机制之一,具体机制有待功能实验验证。

关键词: bta-miR-19b, BCL3, 双荧光素酶报告基因实验, 牛初乳, 免疫调控

Abstract: [Objective] To validate the targeting relationship between bta-miR-19b and the B-cell CLL/lymphoma 3 （BCL3） gene and to elucidate the molecular function and regulatory mechanism by which bta-miR-19b mediates immune regulation in bovine colostrum. [Methods] The intersection of TargetScan and miRanda predictions was used to screen the immune-related target genes of bta-miR-19b, and the minimum free energy （MFE） of binding between bta-miR-19b and immune-related candidate genes IL7R, EGFR, TNF, BCL3, NME7, MAPK8 and MAP3K1 was predicted using the RNAhybrid website. The wild-type （WT） and mutant （MT） fragments were synthesized using the predicted binding site of the 3′UTR of the BCL3 gene as template to construct pmirGLO luciferase reporter vectors. The bta-miR-19b mimics （chemically synthesized double-stranded RNA molecules that mimic endogenous miRNA functions） and NC control （NC-mimics） were synthesized and co-transfected with the reporter vector into 293T cells for target validation. The biological processes and key pathways of bta-miR-19b-BCL3 axis in the immune regulation of bovine colostrum were screened by bioinformatics methods. [Results] The binding free energy of bta-miR-19b with BCL3 was the lowest, suggesting that there was a potential binding of BCL3. The results of dual luciferase reporter gene assay confirmed that bta-miR-19b could specifically bind to the 3′UTR region of the BCL3 gene. The bta-miR-19b-BCL3 regulatory axis was enriched in 42 GO terms, of which 24 were immune-related biological processes. The KEGG pathways involving the bta-miR-19b-BCL3 regulatory axis included tumor necrosis factor TNF signaling pathway and C-type lectin receptor signaling pathway. [Conclusion] bta-miR-19b can target the 3′UTR of BCL3. Given that BCL3 is a regulatory factor of the NF-κB signaling pathway, it is speculated that the bta-miR-19b-BCL3-NF-κB-TNF regulatory axis may be one of the core molecular mechanisms for bovine colostrum to exert immune function, and the specific mechanism needs to be validated by functional experiments.

Key words: bta-miR-19b, BCL3, double luciferase reporter gene assay, bovine colostrum, immune regulation

中图分类号:

S823.2
Q782

孔繁霞, 王高峰, 赵濛, 吴美灵, 王乐乐. bta-miR-19b靶向BCL3的验证及其在牛初乳免疫调控中的功能分析[J]. 畜牧与饲料科学, 2026, 47(3): 23-29.

KONG Fanxia, WANG Gaofeng, ZHAO Meng, WU Meiling, WANG Lele. Validation of bta-miR-19b Targeting BCL3 and Its Functional Analysis in Bovine Colostrum Immune Regulation[J]. Animal Husbandry and Feed Science, 2026, 47(3): 23-29.

参考文献

[1] 王大胜. 奶牛免疫应答性状对犊牛被动免疫和生长健康的影响[D].扬州:扬州大学,2025.
[2] ZHANG C, ZHENG J, HAN X T, et al.Bovine colostrum miR-30a-5p targets the NF-κB signaling pathway to alleviate inflammation in intestinal epithelial cells[J].Journal of Agricultural and Food Chemistry,2024,72(16):9164-9178.
[3] LIN D H, ELCAVAGE L E, KHALIZEVA E, et al.mRNA 3′ UTRs direct microRNA degradation to participate in imprinted gene networks and regulate growth[J].Genes and Development,2026,40(7/8):586-603.
[4] SONG X W, NIU L Z, FAN X X, et al.Overexpression of Rtl1 via synonymous mutation silencing miRNA target site drives skeletal muscle hypertrophy and inflammation in mice[J].BMC Biology,2026,24:67.
[5] WANG L L, WANG G F, ZHAO M, et al.Analysis of immune-related microRNAs in Bovine milk exosomes and construction of their regulatory networks[J].Journal of Applied Animal Research,2025,53:2512724.
[6] WANG V Y, LI Y D, KIM D, et al.Bcl3 phosphorylation by Akt, Erk2, and IKK is required for its transcriptional activity[J].Molecular Cell,2017,67(3):484-497.
[7] CAPECE D, VERZELLA D, FLATI I, et al.NF-κB:blending metabolism, immunity, and inflammation[J].Trends in Immunology,2022,43(9):757-775.
[8] 高静涛. Bcl3在对乙酰氨基酚诱导的急性肝损伤中的作用与机制研究[D].乌鲁木齐:新疆医科大学,2025.
[9] LIU H, ZENG L, YANG Y, et al.Bcl-3:a double-edged sword in immune cells and inflammation[J].Frontiers in Immunology,2022,13:847699.
[10] 秦建宁. miR-223-3p靶向Bcl3激活NF-κB P50促进糖尿病心肌病炎症和重构[D].衡阳:南华大学,2024.
[11] 王乐乐,王高峰,张宇琪,等.牛乳外泌体bta-miR-19b、bta-miR-186调控免疫反应的生物信息学研究[J].内蒙古医科大学学报,2025,47(4):337-342,354.
[12] 王乐乐,王高峰,赵濛.基于MAPK信号通路探讨牛乳及其外泌体关键miRNA的免疫调节机制[J].畜牧与饲料科学,2025,46(3):9-16.
[13] AVILA-BONILLA R G, SALAS-BENITO J S. Computational screening to predict microRNA targets in the flavivirus 3′ UTR genome:an approach for antiviral development[J].International Journal of Molecular Sciences,2024,25(18):10135.
[14] LI Y H, YANG J, WANG X P, et al.Transcriptome analysis reveals the regulation of miR-19b on inflammation in bovine mammary epithelial cells[J].Microbial Pathogenesis,2024,197:107082.
[15] HERRINGTON F, NIBBS R.Regulation of the adaptive immune response by the IκB family protein bcl-3[J].Cells,2016,5(2):14.
[16] 李雪,张捷,周鹏.免疫球蛋白对儿童免疫发育的影响[J].中国乳品工业,2025,53(11):22-30,68.
[17] 杨叶. 牛初乳免疫调节机制及其应用研究进展[J].中国乳业,2026(1):109-117.
[18] ALI A H, NAJJAR Z, LIU S Q, et al.microRNAs from colostrum and mature cow milk:high-throughput sequencing and functional enrichment[J].Food Chemistry:Molecular Sciences,2026,12:100337.
[19] WESSELLS J, BAER M, YOUNG H A, et al.BCL-3 and NF-κB p50 attenuate lipopolysaccharide-induced inflammatory responses in macrophages[J].Journal of Biological Chemistry,2004,279(48):49995-50003.
[20] HOWELL J A, BIDWELL G L III. Targeting the NF-κB pathway for therapy of ischemic stroke[J].Therapeutic Delivery,2020,11(2):113-123.
[21] KUWATA H, WATANABE Y, MIYOSHI H, et al.IL-10-inducible Bcl-3 negatively regulates LPS-induced TNF-α production in macrophages[J].Blood,2003,102(12):4123-4129.
[22] KREISEL D, SUGIMOTO S, TIETJENS J, et al.Bcl3 prevents acute inflammatory lung injury in mice by restraining emergency granulopoiesis[J].Journal of Clinical Investigation,2011,121(1):265-276.
[23] 彭若青,张少言,陈佳骏,等.肿瘤坏死因子-α在分枝杆菌肉芽肿免疫中的研究进展[J].中国现代医学杂志,2025,35(20):60-65.
[24] GHOSH S, IACUCCI M.Diverse immune effects of bovine colostrum and benefits in human health and disease[J].Nutrients,2021,13(11):3798.
[25] SIENKIEWICZ M, SZYMAŃSKA P, FICHNA J. Supplementation of bovine colostrum in inflammatory bowel disease:benefits and contraindications[J].Advances in Nutrition,2021,12(2):533-545.
[26] 杨箭,李宇航,任倩倩,等.Bta-miR-19b在LPS诱导的奶牛(Bos taurus)乳腺上皮细胞中的表达及其生物信息学分析[J].基因组学与应用生物学,2022,41(5):993-1005.
[27] WANG Y J, WANG L, WEN X, et al.NF-κB signaling in skin aging[J].Mechanisms of Ageing and Development,2019,184:111160.

bta-miR-19b靶向BCL3的验证及其在牛初乳免疫调控中的功能分析

Validation of bta-miR-19b Targeting BCL3 and Its Functional Analysis in Bovine Colostrum Immune Regulation

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 4

编辑推荐

Metrics

本文评价

[1]	王乐乐, 王高峰, 赵濛. 基于MAPK信号通路探讨牛乳及其外泌体关键miRNA的免疫调节机制[J]. 畜牧与饲料科学, 2025, 46(3): 9-16.
[2]	李晓奇, 王凤武, 韩健健. 间充质干细胞发挥抗菌作用的机制研究进展[J]. 畜牧与饲料科学, 2022, 43(2): 23-27.
[3]	韩永霞,刘文,韩海霞. 牛初乳中的功能性组分及保健功效[J]. 畜牧与饲料科学, 2009, 30(3): 37-37.
[4]	马燕芬,陈志伟,侯先志. 牛初乳的生物活性物质及其功能[J]. 畜牧与饲料科学, 2005, 26(6): 57-57.