Circ_0007706 downregulation ameliorates neonatal hypoxic ischemic encephalopathy via regulating the miR-579-3p/TRAF6 axis

Jinguang Wang; Minmin Hua; Huixin Li; Dan Xu; Fangfang Li; Falin Xu

doi:10.1016/j.brainresbull.2023.01.013

Circ_0007706 downregulation ameliorates neonatal hypoxic ischemic encephalopathy via regulating the miR-579-3p/TRAF6 axis

Brain Res Bull. 2023 Mar:194:90-99. doi: 10.1016/j.brainresbull.2023.01.013. Epub 2023 Jan 30.

Authors

Jinguang Wang¹, Minmin Hua², Huixin Li³, Dan Xu⁴, Fangfang Li⁵, Falin Xu⁶

Affiliations

¹ Department of Neonatology, The Third Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China. Electronic address: 437537002@qq.com.
² Department of Neonatology, The Third Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China. Electronic address: 952756495@qq.com.
³ Department of Anesthesiology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China. Electronic address: Lihuixin0321@163.com.
⁴ Department of Neurology, The Third Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China. Electronic address: 723932168@qq.com.
⁵ Department of Respiratory Medicine, The Third Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China. Electronic address: Ff880319@163.com.
⁶ Department of Neonatology, The Third Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China. Electronic address: zdsfy123456@163.com.

PMID: 36720318
DOI: 10.1016/j.brainresbull.2023.01.013

Abstract

Background: Neonatal hypoxic ischemic encephalopathy (HIE) is a main factor of neonatal death and permanent neurologic deficit. This study sought to investigate the functional role of hsa_circ_0007706 (circ_0007706) in modulating neonatal HIE.

Methods: In vitro HIE cell model was established in hBMVECs under the condition of oxygen‑glucose deprivation/reperfusion (OGD/R) treatment. qRT-PCR analysis was utilized for detecting the level of circ_0007706, microRNA-579-3p (miR-579-3p) and TNF receptor-associated factor 6 (TRAF6). RNase R treatment and Oligo (dT) ₁₈ primers were employed to verify the features of circ_0007706, and nucleocytoplasmic separation was conducted for determining the location of circ_0007706. CCK-8 assay, EdU assay, and flow cytometry were carried out to measure cell proliferation and apoptosis, respectively. The protein expression of Bax, Bcl-2 and TRAF6 was detected using western blot. Meanwhile, the levels of the pro-inflammatory factors were determined via ELISA. SOD activity and MDA level were assessed via the respective kits. Besides, dual-luciferase reporter assay and RNA pull-down were used to identify the association between miR-579-3p and circ_0007706 or TRAF6.

Results: Circ_0007706 was elevated in HIE newborns and OGD/R cell model. Knockdown of circ_0007706 greatly alleviated OGD/R-induced injury, inflammatory response and oxidative stress. We found that miR-579-3p was a direct target of circ_0007706, and miR-579-3p inhibitor could reverse the impact of circ_0007706 knockdown on OGD/R-caused cell damage in hBMVECs. In addition, miR-579-3p directly interacted with TRAF6, and the protective effects of miR-579-3p on OGD/R-induced injury in hBMVECs were harbored by TRAF6 overexpression. Our data indicated that circ_0007706 knockdown could downregulate the expression of TRAF6 by sponging miR-579-3p in OGD/R-treated hBMVECs.

Conclusion: This study demonstrated that circ_0007706 knockdown assuaged HIE-induced injury by decreasing TRAF6 expression via targeting miR-579-3p.

Keywords: Circ_0007706; Neonatal hypoxic ischemic encephalopathy; TRAF6; miR-579-3p.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Apoptosis
Cell Proliferation / genetics
Down-Regulation
Glucose
Humans
Hypoxia-Ischemia, Brain* / genetics
Infant, Newborn
MicroRNAs* / genetics
TNF Receptor-Associated Factor 6 / genetics

Substances

TNF Receptor-Associated Factor 6
Glucose
MicroRNAs
MIRN579 microRNA, human