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