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During the congress, E-Posters will be accessible to all participants on the congress website 24/7, as well as in the E-poster stations in the congress center.
Preparing your E-Poster
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E-Poster Submission Deadline
Please prepare and upload your E-Poster no later than March 14, 2026 11.59PM CET. After this date, you will no longer be able to prepare and upload your E-poster and it will not be displayed and accessible on the congress website.
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Abstract titles should be brief and reflect the content of the abstract.
Chronic kidney disease has developed into a critical public health problem around the world, the prevalence of CKD is nearly 15% worldwide, and China is the world’s leading country with 132 million people suffering from the disease. As CKD progresses to end-stage, the risk of death and systemic complications increases dramatically placing a heavy burden on patients and society. Interstitial fibrosis is an important pathological process and a common pathway in the progression of various kidney diseases to end-stage, and improving interstitial fibrosis can effectively slow the progression of CKD. In recent years, the role of circadian rhythms and clock genes in kidney injury has gradually received more attention, but most of the existing studies focus on core clock genes. In contrast, D-site binding protein (DBP), as an important clock output gene, can more directly regulate a variety of physiopathological processes and is an ideal target for precise disease intervention. However, no study has investigated its role in renal interstitial fibrosis. Based on the above, the present study aimed to clarify the expression characteristics, function and specific molecular mechanism of DBP in renal interstitial fibrosis. From the perspective of clock genes, this study will provide a new perspective and idea on the pathogenesis of interstitial fibrosis, as well as a theoretical and laboratory foundation for the preventing and treating of renal fibrosis.
1) Unilateral ureteral obstruction (UUO) was used to induce the renal interstitial fibrosis model in C57BL/6 male mice. The kidney tissues of the two groups of mice were also subjected to high-throughput RNA-Seq sequencing and analysis, and the clock genes with the most significant differences were selected and their expression in the kidney tissues was verified by WB and real-time quantitative PCR (RT-qPCR). In addition, HK2 cells were cultivated in vitro and the cellular fibrosis model was induced with 10 ng/ml TGF-β1, and WB, RT-qPCR and immunofluorescence staining were also used to determine the expression of DBP in HK2 cells. Meanwhile, 10 cases each of kidney tissues from CKD patients and paracancerous kidney tissues from renal tumour patients were collected, and their kidney tissue sections were subjected to Masson's and Sirius Scarlet staining to assess the degree of fibrosis, and the expression of α-SMA, vimentin and DBP were detected by immunohistochemical staining.
2) DBP systemic knockout (DBP KO) mice were generated using CRISPR/Cas9 technology and subjected to UUO surgery and intraperitoneal injection of folic acid to induce a model of renal interstitial fibrosis. DBP overexpression mice (AAV-DBP) and control mice were generated by tail vein injection of mouse DBP-AAV9 adenoassociated viruses and the renal interstitial fibrosis model was established by UUO surgery. Serum creatinine, blood urea nitrogen and cystatin C were measured to assess renal function. Renal tissue sections were stained with HE staining, Masson staining and Wolf Scarlet staining to assess the extent of tubular damage and interstitial fibrosis. The expression of fibrotic indexes (α-SMA, vimentin， E-cadherin，Tgf-β, Mmp9 and Col1al) and inflammatory indexes (Il-1β, Il-6 and Tnf-α) in mouse kidneys were detected by WB ， RT-qPCR, immunofluorescence and immunohistochemical staining. The siRNA and overexpression plasmid of DBP were respectively constructed and transfected into TGF-β1-induced HK2 cells, and the expression of DBP and α-SMA were detected by WB and RT-qPCR.
3) The results of RNA sequencing of mouse kidney tissue, transcriptome datasets GSE54650 and PRDB7796 were intersected to preliminarily identify downstream target genes that might be regulated by DBP. WB and RT-qPCR were used to detect the expression of target genes and their downstream factors in the kidneys of DBP systemic knockout and DBP overexpressing UUO mice. DBP targeting to the promoter regions of downstream target genes as well as the specific binding sites were also clarified using a dual luciferase reporter gene assay. Finally, rescue experiments were performed to inhibit downstream target genes in DBP-overexpressing UUO mice, and serum renal function indices were determined, and the degree of tubular damage and interstitial fibrosis was assessed by HE staining, Masson staining and Wolf Scarlet staining; immunofluorescence staining, WB and RT-qPCR were used to detect fibrosis-related indices and changes in factors downstream of the target genes.
1) The results of high-throughput RNA-Seq sequencing showed that UUO mice kidney tissues contained abnormal expression of various clock genes, the most significant of which was up-regulation of DBP, and the protein and mRNA levels were significantly increased which was further confirmed by WB and RT-qPCR. In vitro, the protein and mRNA expression of α-SMA and DBP in HK2 cells were significantly increased after TGF-β1 intervention, and the intensity of DBP immunofluorescence staining was significantly increased. In clinical samples, DBP expression in kidney tissue was markedly higher in CKD patients than in controls, and was more abundant in the renal tubules, and the results from the Nakagawa database were also consistent.
2) In vivo, compared with the control group, systemic knockdown of DBP reduced renal function indicators (CRE, BUN, CysC), attenuated renal tubular injury and interstitial collagen deposition, and significantly decreased the expression of fibrosis indicators (α-SMA, Tgf-β, Mmp9 and Col1al) and inflammatory indicators (Il-1β, Il-6, and Tnf-α) in renal tissues, Consistently, systemic knockdown of DBP also significantly ameliorated folate-induced renal interstitial fibrosis at multiple levels, including functional, tissue and molecular levels. Conversely, overexpression of DBP in UUO mice resulted in further deterioration of renal function, increased tubular injury and interstitial fibrosis, and significant up-regulation of fibrosis indices and renal inflammation indices in mouse kidney. In vitro, knockdown of DBP significantly reduced TGF-β1-induced α-SMA elevation in HK2 cells, and conversely, overexpression of DBP further promoted TGF-β1-induced α-SMA upregulation in HK2 cells.
3) The GSE54650 dataset screened 2351 genes that were rhythmically expressed in kidney tissue; the PRDB7796 dataset screened 5471 DBP-directly bindable target genes; high-throughput RNA-Seq sequencing of mouse kidney tissue identified 80 genes that were significantly up-regulated in the WT+UUO group and down-regulated in the DBP KO+UUO group. The intersection of the above three datasets shows that TGFBR2 is a downstream target gene directly regulated by DBP. WB and RT-qPCR results further indicated that the expression of TGFBR2 and its downstream factor p-Smad2/3 was significantly decreased in the kidney tissues of UUO mice after DBP knockdown and, conversely, was significantly increased after DBP overexpression. Meanwhile, the result of the dual luciferase assay showed that the '-939 ~ -948，ATTATCCAAT' site in the promoter region of TGFBR2 is the target binding site of DBP, which activates its transcription. Rescue experiments further demonstrated that ITD-1, a selective inhibitor of TGFBR2, significantly attenuated the promoting effect of DBP overexpression on renal fibrosis in UUO mice.
Clock genes DBP is aberrantly increased expressed in the state of renal interstitial fibrosis. It can directly target the '-939 ~ -948，ATTATCCAAT' site in the promoter region of TGFBR2 and activate its transcription, which promotes the phosphorylation of the downstream factor Smad2/3 and then exerts a pro-renal interstitial fibrosis effect.
