天津医药

天津医药 ›› 2019, Vol. 47 ›› Issue (12): 1224-1229.doi: 10.11958/20191174

• 实验研究 • 上一篇    下一篇

转录因子D位点结合蛋白对原代大鼠系膜细胞中 免疫炎症因子的调控作用

蒋红利#，程鑫#，史珂慧，何荃，魏萌，王萌，陈蕾△   

  1. 西安交通大学第一附属医院血液净化科（邮编710061）
  • 收稿日期:2019-04-16 修回日期:2019-08-12 出版日期:2019-12-15 发布日期:2019-12-15
  • 通讯作者: 陈蕾 E-mail:445046614@qq.com
  • 作者简介:蒋红利（1966），女，主任医师，博士，主要从事肾脏病学与血液净化学研究；程鑫（1989），女，硕士，医师，主要从事肾脏病学与血 液净化学研究
  • 基金资助:
    西安交通大学第一附属医院青年创新基金;西安市科技计划项目

The regulatory role of transcription factor D-site binding protein on expression of immune and inflammatory factors in primary rat mesangial cells

 JIANG Hong-li#, CHENG Xin#, SHI Ke-hui, HE Quan, WEI Meng, WANG Meng, CHEN Lei△   

  1. Department of Dialysis, the First Affiliated Hospital of Xi’an Jiaotong University, Xi’an 710061, China
  • Received:2019-04-16 Revised:2019-08-12 Published:2019-12-15 Online:2019-12-15
  • Contact: Lei Chen E-mail:445046614@qq.com
  • Supported by:
     

PDF (PC)

3390

摘要： 目的 通过基因表达谱分析，探讨并验证转录因子D位点结合蛋白（DBP）对原代大鼠系膜细胞（RMCs）中 免疫炎症因子的调控作用。方法 从8周龄SD大鼠中分离、培养并鉴定原代RMCs，利用小干扰RNA技术敲低其 DBP表达，进而行RNA-seq测序分析；筛选差异表达基因（DEGs），并对DEGs行Gene Ontology（GO）注释及KEGG信 号通路分析；采用qPCR验证DBP低表达的RMCs中免疫炎症因子mRNA表达水平；利用质粒转染技术获得DBP过表 达RMCs并采用qPCR检测DBP过表达对免疫炎症因子mRNA表达的影响。结果 成功分离、培养原代RMCs，并获 得DBP低表达的原代RMCs。RNA-seq测序结果显示，与野生原代RMCs相比，DBP低表达的原代RMCs中共有267 个DEGs（上调84个，下调183个）。GO注释及KEGG信号通路分析表明，DEGs主要富集在免疫炎症相关的生物学过程 及信号通路。原代RMCs中，DBP敲低可明显下调免疫炎症因子C-C模体趋化因子配体2（CCL2）、C-X-C模体趋化因 子（CXCL）10、CXCL1、肿瘤坏死因子（TNF）-α、白细胞介素（IL）-1、IL-6 mRNA表达，DBP过表达则可明显上调上述免 疫炎症因子mRNA表达。结论 原代RMCs中，DBP可通过调控免疫炎症因子表达参与免疫炎症反应。

关键词: 肾小球系膜细胞, 肾小球肾炎, 膜增生性, 基因表达谱, D位点结合蛋白, 免疫炎症因子, 系膜增殖性肾炎

Abstract: Objective To investigate and verify the regulatory role of D-site binding protein (DBP) on the expression of immune and inflammatory factors in primary rat mesangial cells (RMCs) through gene expression profile analysis. Methods The primary RMCs were isolated from 8-week-old SD rats, then cultured for next study. The DBP expression was knocked down by small interfering RNA in primary RMCs, which were used for RNA-seq analysis. Based on the results of RNA-seq analysis, differentially expressed genes (DEGs) were screened out. Gene Ontology (GO) annotation terms and KEGG signaling pathway analysis were conducted for DEGs. The mRNA expression level of immune and inflammatory factors in DBP-knockdown primary RMCs were detected by qPCR. Furthermore, plasmid transfection was used to obtain DBP-overexpressed primary RMCs, and the effect of DBP overexpression on mRNA expression of immune and inflammatory factors was detected by qPCR. Results The primary RMCs were isolated and cultured successfully, and the primary RMCs with DBP knockdown was obtained for RNA-seq sequencing. Compared with the wild primary RMCs, 267 DEGs were found in the primary RMCs with DBP knockdown (84 DEGs up-regulated, 183 DEGs down-regulated). GO annotation terms and KEGG signaling pathway analysis showed that DEGs were mainly enriched in biological processes and signaling pathways related to immune and inflammatory response. In primary RMCs, the knockdown of DBP significantly decreased the mRNA expression of C-C motif chemokine ligand 2 (CCL2), C-X-C motif chemokine 10 (CXCL10), C-X-C motif chemokine 1 (CXCL1), tumor necrosis factor alpha (TNF -α), interleukin 1(IL-1) and interleukin 6 (IL-6), while the overexpression of DBP induced their mRNA expression. Conclusion In primary RMCs, transcription factor DBP regulates the expression of immune and inflammatory factors.

Key words: mesangial cells, glomerulonephritis, membranoproliferative, gene expression profiling, D-site binding protein, immune and inflammatory factors, mesangial proliferative glomerulonephritis

中图分类号: