Knockdown of microRNA-96-5p resists oxidative stress-induced apoptosis in nucleus pulposus cells

Ji Wang; Ziming Xia; Zhengyi Su; Hong Xue; Chengjun Huang; Ming Su

Knockdown of microRNA-96-5p resists oxidative stress-induced apoptosis in nucleus pulposus cells

Am J Transl Res. 2023 Jul 15;15(7):4912-4921. eCollection 2023.

Authors

Ji Wang¹, Ziming Xia¹, Zhengyi Su¹, Hong Xue¹, Chengjun Huang¹, Ming Su²

Affiliations

¹ Department of Orthopedics, Liuzhou Traditional Chinese Medical Hospital of Guangxi University of Chinese Medicine Liuzhou, Guangxi, P. R. China.
² Department of Pain, Liuzhou Traditional Chinese Medical Hospital of Guangxi University of Chinese Medicine Liuzhou, Guangxi, P. R. China.

PMID: 37560214
PMCID: PMC10408537

Abstract

Background: Intervertebral disc degeneration (IVDD) often leads to low back pain, which severely affects people's quality of life. Oxidative stress (OS) can accelerate nucleus pulposus cell (NPCs) senescence and apoptosis. Exploring the mechanism underlying OS-induced apoptosis is of utmost importance to aid in the development of IVDD treatment.

Methods: In the current study, we tested the function of microRNA-96-5p in H₂O₂-treated NPCs. Apoptosis and mitophagy-related proteins were examined by western blot. Reactive oxygen species (ROS) generation, mitochondrial membrane potential, and apoptosis of NPCs were evaluated by flow cytometry. A luciferase reporter assay was conducted to confirm the interaction between microRNA-96-5p and Forkhead Box Protein O1 (FOXO1).

Results: H₂O₂ treatment enhanced apoptosis in NPCs and upregulated the microRNA-96-5p expression. It was shown that knockdown of microRNA-96-5p attenuated H₂O₂-induced OS and apoptosis. FOXO1 is a direct target of microRNA-96-5p, and knockdown of microRNA-96-5p enhanced PINK1/Parkin-mediated mitophagy by up-regulating FOXO1.

Conclusions: Collectively, knockdown of microRNA-96-5p enhanced PINK1/Parkin-mediated mitophagy by up-regulating FOXO1. Our results facilitate the understanding of the role of microRNA-96-5p in IVDD and the mechanism of H₂O₂-induced oxidative damage.

Keywords: FOXO1; MicroRNA-96-5p; intervertebral disc degeneration; mitophagy; nucleus pulposus cells.