Exhausting circ_0136474 and Restoring miR-766-3p Attenuate Chondrocyte Oxidative Injury in IL-1β-Induced Osteoarthritis Progression Through Regulating DNMT3A

Abstract

Circular RNA circ_0136474 is a new contributor of human osteoarthritis (OA) by suppressing chondrocyte proliferation. However, its role and mechanism in OA chondrocyte injury remain ill defined. Herein, we performed real-time quantitative PCR to detect RNA expression of circ_0136474, microRNA (miR)-766-3p, and DNA methyltransferase 3A (DNMT3A) and utilized Western blotting to measure protein expression of DNMT3A, matrix metalloproteinase-1 (MMP1), MMP13, collagen II, proliferating cell nuclear antigen (PCNA) and B cell lymphoma (Bcl)-2, and Bcl-2-associated X protein (Bax). Direct interaction between miR-766-3p and circ_0136474 or DNMT3A was confirmed by bioinformatics algorithms, dual-luciferase reporter assay, and RNA immunoprecipitation. Functional experiments including cell counting kit-8 assay, flow cytometry, and special assay kits were employed to measure oxidative injury in interleukin (IL)-1β-induced OA-like chondrocytes. First, IL-1β administration induced cell viability inhibition, collagen II suppression, and promotion of MMP1 and MMP13 in human chondrocyte CHON-001 cells. Expression of circ_0136474 and DNMT3A was upregulated, and miR-766-3p was downregulated in human OA cartilages and IL-1β-induced CHON-001 cells. Functionally, both blocking circ_0136474 and upregulating miR-766-3p could rescue cell viability and levels of PCNA, Bcl-2, reduced glutathione (GSH), and total superoxide dismutase (SOD), and attenuate apoptosis rate and levels of Bax, reactive oxygen species (ROS), and lipid peroxidation malondialdehyde (MDA). Mechanically, circ_0136474 served as miR-766-3p sponge to govern miR-766-3p-targeted DNMT3A expression. Accidently, restoring DNMT3A counteracted the miR-766-3p upregulation role, and silencing miR-766-3p weakened circ_0136474 knockdown effect in IL-1β-induced CHON-001 cells. In conclusion, exhausting circ_0136474 could mitigate OA chondrocyte oxidative injury through regulating miR-766-3p/DNMT3A axis.

Circular RNA hsa_circ_0005567 overexpression promotes M2 type macrophage polarization through miR-492/SOCS2 axis to inhibit osteoarthritis progression

“…CircRNA-associated ceRNA network construction reveals the circRNAs involved in the progression of osteoarthritis. Such as has_circ_0136474, which regulates IL-1β-Induced chondrocyte injury through miR-766-3p/DNMT3A Signaling Pathway; Circ-BRWD1, which contributes to osteoarthritis development through the modulation of miR-1277/TRAF6 axis [ 27 , 28 ]. Using RNA pull-down and dual luciferase reporter gene assay, we have demonstrated that hsa_circ_0005567 could regulate SOCS2 expression by miR-492.…”

miR‐423‐3p activates FAK signaling pathway to drive EMT process and tumor growth in lung adenocarcinoma through targeting CYBRD1

“…MicroRNA‐423‐3p (miR‐423‐3p) belongs to microRNAs, which are defined as small noncoding RNA molecules with single strand. 13 Increasingly, studies have uncovered the functional role of microRNAs in various diseases, such as ischemia‐reperfusion injury, 14 human osteoarthritis, 15 and psoriasis. 16 In addition, the research reports about microRNAs have mainly focused on its regulation of multiple cancers, including osteosarcoma, 17 melanoma, 18 and colorectal cancer.…”

The Role Played by Ferroptosis in Osteoarthritis: Evidence Based on Iron Dyshomeostasis and Lipid Peroxidation

“…Zuo et al [ 97 ] and Yin et al [ 98 ] found increased intracellular ROS and decreased GPX levels in the same model cells. Zhu et al [ 99 ] reported increased intracellular ROS and MDA levels and decreased expression of GSH in IL-1β-induced OA-like chondrocytes and showed that circ_0136474 activity or upregulated miR-766-3p expression attenuated oxidative injury. Wang et al [ 100 ] reported increased intracellular ROS and decreased expression of GSH in TNF-α-treated chondrocytes.…”