Abstract
Diabetic nephropathy (DN) has been identified as the major cause of end-stage renal disease (ESRD) in most developed countries. MicroRNA-770-5p depletion could repress high glucose (HG)-triggered apoptosis in podocytes, and downregulation of E2F transcription factor 3 (E2F3) could facilitate podocyte injury. Nevertheless, whether E2F3 is involved in miR-770-5p knockdown-mediated improvement of DN is still unclear. The expression levels of miR-770-5p and E2F3 were detected in HG-treated podocytes by RT-qPCR. The expression levels of E2F3, apoptosis-related proteins Bcl-2 related X protein (Bax), B-cell lymphoma-2 (Bcl-2), Bad, apoptotic peptidase activating factor 1 (APAF1), C-caspase3, C-caspase7, and C-caspase9 were detected by western blot assay. The effects of miR-770-5p and E2F3 on HG-treated podocytes proliferation and apoptosis were detected by CCK-8 and flow cytometry assays. The interaction between miR-770-5p and E2F3 was predicted by Targetscan, and then verified by the dual-luciferase reporter assay. MiR-770-5p was upregulated and E2F3 was downregulated in HG-treated podocytes. MiR-770-5p inhibited proliferation and promoted apoptosis and E2F3 promoted proliferation and suppressed apoptosis in HG-treated podocytes. E2F3 is a target gene of miR-770-5p and it partially abolished the effect of miR-770-5p in HG-triggered proliferation and apoptosis of podocytes. MiR-770-5p deficiency blocked HG-induced APAF1/caspase9 pathway via targeting E2F3 in podocytes. We firstly confirmed that E2F3 was a target of miR-770-5p in podocytes. These findings suggested that miR-770-5p expedited podocyte injury by targeting E2F3, and the miR-770-5p/E2F3 axis might represent a pathological mechanism of DN progression.
Highlights
Diabetic nephropathy (DN) is a type of microvascular complication in diabetes, and has been identified as the major cause of end-stage renal disease (ESRD) in most developed countries [1]
Podocytes were treated with various doses of glucose. miR-770-5p expression was highly expressed in podocytes treated with 30 mM glucose relative to cells treated with 5 and 15 mM glucose (Figure 1A)
30 mM glucose-treated podocytes were cultured in a time-dependent method. miR-770-5p was markedly upregulated at 24 and 48 h compared to 0 h (Figure 1B)
Summary
Diabetic nephropathy (DN) is a type of microvascular complication in diabetes, and has been identified as the major cause of end-stage renal disease (ESRD) in most developed countries [1]. 40% of patients with diabetes develop DN, and the diabetes-related deaths in adults were more than 0.39 million only in 2010 [1,2]. With a rapidly increasing incidence, diabetes has become an important public health concern [3]. A class of highly differentiated glomerular epithelial cell, exert the vital function of maintaining the integrity of the glomerular filtration barrier [4,5]. Previous studies showed that the injury and loss of podocytes is strictly related to the early pathological mechanism of DN [6]. Despite the substantial progress in therapeutic methods, it remains imperative to further explore podocyte-based therapies that can prevent or cure DN
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callMore From: Brazilian Journal of Medical and Biological Research
Disclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
