Abstract: SA-PO134
MiR-874/ADAM19 Mediates Macrophage Activation and Renal Fibrosis After AKI
Session Information
- AKI: Mechanisms - AKI-CKD Transition
November 09, 2019 | Location: Exhibit Hall, Walter E. Washington Convention Center
Abstract Time: 10:00 AM - 12:00 PM
Category: Acute Kidney Injury
- 103 AKI: Mechanisms
Authors
- Wang, Junni, First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, Zhejiang, China
- Nie, Wanyun, First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, Zhejiang, China
- Xie, Xishao, First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, Zhejiang, China
- Han, Fei, First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, Zhejiang, China
Background
The pathogenesis of CKD following AKI is not fully investigated.
Methods
We established a mouse AC (AKI to CKD) model caused by ischemia/reperfusion(I/R), and identified miR-874 downregulated in fibrotic kidneys of both AC model and UUO model by RNA-Seq. Then we used human patient samples as well as animal and cell models to investigate how miR-874 regulates renal fibrosis after I/R injury.
Results
MiR-874 was reduced at different time point after I/R and UUO. In vitro, miR-874 was downregulated in HK2 cells treated with TGF-β1. Moreover, miR-874 level of peripheral mononuclear cells was lower in IgA nephropathy (IgAN) patients with proliferative sclerosing glomerulonephritis than those in pathological stage M0E0S0T0(p<0.01). In vitro, transient transfection of miR-874 inhibitor in HK2 cells induced the increase of mesenchymal makers, and transfection of miR-874 mimic in HK2 cells treated with TGF-β1 could alleviate EMT compared with negative control. Overexpression of human miR-874 into AC and UUO mice led to alleviated renal fibrosis with decreased expression level of Acta2, Col1a1, Fn1, chemokines including CCL2/CCL5, and ADAM19, a target gene of miR-874 verified by luciferase microRNA target reporter assay. F4/80 staining was also junior in miR-874 mice compared with negative control. In vitro, transfection of miR-874 mimic in mouse macrophage cell line Raw264.7 stimulated with LPS downregulated the expression of CCL2/CCL5. Then we focused on the biological function of ADAM19 expression towards renal fibrosis. ADAM19 was induced in both UUO mice and AC mice at different time point. Transfection of adenovirus carrying human ADAM19 into HK2 cells, ADAM19 could induce the increase of mesenchymal makers and inflammatory factors and decrease of epithelial markers. Overexpression of ADAM19 directly induced fibrotic changes in vivo. The expression of CCL2/CCL5 and F4/80 staining were also upregulated in ADAM19 mice compared with negative control. In vitro, transfection of adenovirus carrying human ADAM19 into Raw264.7 cells increased nitric oxide synthase 2 (NOS2), chemokines and proinflammatory cytokines such as interleukin-1β (IL-1β) at both mRNA and protein levels.
Conclusion
Our results suggest miR-874/ADAM19 could mediate renal fibrosis through regulating renal tubular epithelial cell injury and macrophage activation.