MiR-182-5p and miR-378a-3p regulate ferroptosis in I/R-induced renal injury

Chenguang Ding,Yuxi Qiao,Bo Wang,Jin Zheng,Xiaoming Ding,Yang Li,Meng Dou,Heli Xiang,Puxun Tian,Wujun Xue

doi:10.1038/s41419-020-03135-z

Abstract

Renal tubular cell death is the key factor of the pathogenesis of ischemia/reperfusion (I/R) kidney injury. Ferroptosis is a type of regulated cell death (RCD) found in various diseases. However, the underlying molecular mechanisms related to ferroptosis in renal I/R injury remain unclear. In the present study, we investigated the regulatory role of microRNAs on ferroptosis in I/R-induced renal injury. We established the I/R-induced renal injury model in rats, and H/R induced HK-2 cells injury in vitro. CCK-8 was used to measure cell viability. Fe2+ and ROS levels were assayed to evaluate the activation of ferroptosis. We performed RNA sequencing to profile the miRNAs expression in H/R-induced injury and ferroptosis. Western blot analysis was used to detect the protein expression. qRT-PCR was used to detect the mRNA and miRNA levels in cells and tissues. We further used luciferase reporter assay to verify the direct targeting effect of miRNA. We found that ischemia/reperfusion-induced ferroptosis in rat’s kidney. We identified that miR-182-5p and miR-378a-3p were upregulated in the ferroptosis and H/R-induced injury, and correlates reversely with glutathione peroxidases 4 (GPX4) and solute carrier family 7 member 11 (SLC7A11) expression in renal I/R injury tissues, respectively. In vitro studies showed that miR-182-5p and miR-378a-3p induced ferroptosis in cells. We further found that miR-182-5p and miR-378a-3p regulated the expression of GPX4 and SLC7A11 negatively by directly binding to the 3′UTR of GPX4 and SLC7A11 mRNA. In vivo study showed that silencing miR-182-5p and miR-378a-3p alleviated the I/R-induced renal injury in rats. In conclusion, we demonstrated that I/R induced upregulation of miR-182-5p and miR-378a-3p, leading to activation of ferroptosis in renal injury through downregulation of GPX4 and SLC7A11.

Highlights

Acute renal failure (ARF) is characterized by the sustained loss of kidney function, of which the primary cause is ischemia/reperfusion (I/R) kidney injury[1]
We found that the glutathione peroxidases 4 (GPX4), SLC7A11, FTH1, and Keap[1] were downregulated, and Nrf[2], NOX1, and COX2 were upregulated in the I/R rats’ kidney tissues, indicating the activation of ferroptosis in the I/R-induced kidney injury (Fig. 3F)
Renal tubular cell death induced by I/R was considered as the main cause of ARF19,46

Summary

Introduction

Acute renal failure (ARF) is characterized by the sustained loss of kidney function, of which the primary cause is ischemia/reperfusion (I/R) kidney injury[1]. I/R injury is prone to occur after kidney transplantation, leading to the failure of the transplanted kidney[2,3,4]. It is well recognized that the death of renal tubular cells, induced death of renal tubular cells is still not fully understood[8,9]. Ferroptosis is a type of regulated cell death (RCD), which is different from other forms of cell death characterized by the accumulation of iron, lipid peroxidation, and condensed mitochondrial membrane densities[10]. Ferroptosis has been found to participate in many disease processes, such as cancer development, myocardial infarction, and neurological disease[11,12,13,14].

Methods

Results

Conclusion