miR-125b参与TRAF6影响的人胰腺导管上皮屏障作用

基础医学与临床 ›› 2021, Vol. 41 ›› Issue (9): 1272-1276.

miR-125b参与TRAF6影响的人胰腺导管上皮屏障作用

韦碧薇^1#, 龚雅慧^1,2#, 苏州¹, 杨慧莹¹, 覃蒙斌³, 梁志海^1*

1.广西医科大学第一附属医院消化内科,广西南宁 530021;
2.荆州市中心医院儿科, 湖北荆州 434020;
3.广西医科大学第二附属医院消化内科, 广西南宁 530007

收稿日期:2020-11-09 修回日期:2021-04-30 出版日期:2021-09-05 发布日期:2021-09-02
通讯作者: *ahhai@163.com
作者简介:#对本文有相同贡献
基金资助:
广西自然科学基金(2017GXNSFAA198068);广西自然科学基金青年科学基金(2018GXNSFBA281154);南宁市青秀区科技计划(2019026)

Involvement of miR-125b in human pancreatic ductal epithelial barrier affected by TRAF6

WEI Bi-wei^1#, GONG Ya-hui^1,2#, SU Zhou¹, YANG Hui-ying¹, QIN Meng-bin³, LIANG Zhi-hai^1*

1. Department of Gastroenterology, the First Affiliated Hospital of Guangxi Medical University, Nanning 530021;
2. Department of Pediatrics, Jingzhou Central Hospital, Jingzhou 434020;
3. Department of Gastroenterology, the Second Affiliated Hospital of Guangxi Medical University, Nanning 530007, China

Received:2020-11-09 Revised:2021-04-30 Online:2021-09-05 Published:2021-09-02
Contact: *ahhai@163.com

摘要/Abstract

摘要： 目的探讨肿瘤坏死因子受体相关因子6(TRAF6)对人胰腺导管上皮细胞系HPDE6C7屏障的影响,以及miR-125b与TRAF6的靶向结合关系。方法用重组慢病毒感染上调HPDE6C7细胞的TRAF6表达。Western blot检测核因子κB(NF-κB)通路蛋白和紧密连接蛋白occludin和claudin-1的表达量;Transwell小室法检测单层HPDE6C7细胞的异硫氰酸荧光素-葡聚糖(FITC-D)透过率以评估上皮屏障功能;双荧光素酶报告基因实验检测miR-125b与TRAF6是否存在靶向结合关系。结果经重组慢病毒感染并上调TRAF6基因表达后,HPDE6C7细胞的NF-κB p65、TAK1和TAB1蛋白表达水平上升(P＜0.05),occludin 和claudin-1蛋白表达水平下降(P＜0.05),单层细胞通透率明显上升(P＜0.05);双荧光素酶报告基因实验证实TRAF6与miR-125b靶向结合。结论 TRAF6通过调控NF-κB信号通路影响胰腺导管上皮屏障功能;miR-125b可能通过与TRAF6靶向结合抑制该作用。

关键词: 急性胰腺炎, 胰腺导管上皮屏障, 肿瘤坏死因子受体相关因子6(TRAF6), 核因子κB(NF-κB), miR-125b

Abstract: Objective To investigate the effect of tumor necrosis factor receptor-associated factor 6 (TRAF6) on barrier and targeted binding of miR-125b to TRAF6 in human pancreatic ductal epithelial cell line HPDE6C7. Methods TRAF6 expression HPDE6C7 cells were up-regulated by recombinant lentivirus infection. The protein expression of nuclear factor κB(NF-κB) pathway and tight junction were examined by Western blot, the permeability of monolayer HPDE6C7 cells was measured with Transwell cab to evaluate the epithelial barrier function, and the targeted binding of miR-125b to TRAF6 was identified by dual-luciferase reporter assay. Results After up-regulation of TRAF6 expression by HPDE6C7 cells, the protein level of NF-κB p65, TGFβ-activated kinase 1(TAK1) and TAK1-binding 1 (TAB1) was increased, and the protein level of occludin and claudin-1 was decreased(P＜0.05), the permeability increased significantly (P＜0.05). Dual-luciferase reporter assay confirmed that TRAF6 was the target gene of miR-125b. Conclusions TRAF6 potentially play a role in affecting the pancreatic ductal epithelial barrier by regulating the NF-κB signaling pathway, which may be inhibited by miR-125b targeted binding to TRAF6.

Key words: acute pancreatitis, pancreatic ductal epithelial barrier, tumor necrosis factor receptor-associated factor 6 (TRAF6), nuclear factor κB (NF-κB), miR-125b

中图分类号:

R576

韦碧薇, 龚雅慧, 苏州, 杨慧莹, 覃蒙斌, 梁志海. miR-125b参与TRAF6影响的人胰腺导管上皮屏障作用[J]. 基础医学与临床, 2021, 41(9): 1272-1276.

WEI Bi-wei, GONG Ya-hui, SU Zhou, YANG Hui-ying, QIN Meng-bin, LIANG Zhi-hai. Involvement of miR-125b in human pancreatic ductal epithelial barrier affected by TRAF6[J]. Basic & Clinical Medicine, 2021, 41(9): 1272-1276.

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