Research Paper Volume 15, Issue 23 pp 14457—14472
MiR-203a-3p attenuates apoptosis and pyroptosis of chondrocytes by regulating the MYD88/NF-κB pathway to alleviate osteoarthritis progression
- 1 Zhongshan Hospital of Traditional Chinese Medicine Affiliated to Guangzhou University of Traditional Chinese Medicine, Zhongshan 528401, Guangdong, China
- 2 The Second Clinical College of Guangzhou University of Chinese Medicine, Guangdong Provincial Hospital of Chinese Medicine, Guangzhou 510120, Guangdong China
- 3 Liuyang Hospital of Traditional Chinese Medicine, Liuyang 410300, Hunan, China
Received: August 16, 2023 Accepted: November 20, 2023 Published: December 13, 2023
https://doi.org/10.18632/aging.205373How to Cite
Copyright: © 2023 Chen et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Abstract
Background: Osteoarthritis (OA) is a degenerative joint disease that imposes a significant socioeconomic burden worldwide. Our previous studies revealed a down-regulation of miR-203a-3p in the knee tissues of OA patients. However, the underlying mechanism through which miR-203a-3p mediates the pathological process of OA remains unknown. Thus, we aimed to determine the effects of miR-203a-3p in the progression of OA.
Methods: Rat primary chondrocytes were stimulated with 10 μg/mL lipopolysaccharide (LPS) for 24 hours, followed by transfection with 50 nM miR-203a-3p mimic, inhibitor, and siRNA for MYD88 or consistent negative controls for 48 hours. To evaluate the effects of miR-203a-3p on cartilage matrix degradation, oxidative stress, apoptosis, and pyroptosis in chondrocytes, various techniques such as immunofluorescence staining, biochemical analysis, Western blotting, and the TUNEL staining were utilized. In the rat OA model, all rats were randomly divided into four groups: Sham, OA, OA+Agomir negative control (NC), and OA+Agomir. They received intra-articular injections of 25 nmol miR-203a-3p agomir, agomir NC, or normal saline twice a week for the duration of 8 weeks after OA induction. Immunofluorescence staining was performed to evaluate the effects of miR-203a-3p on cartilage matrix degradation in rats.
Results: MiR-203a-3p was down-regulated in LPS-treated rat chondrocytes and OA cartilage, and directly targeted MYD88. Moreover, miR-203a-3p significantly inhibited LPS-induced cartilage matrix degradation, oxidative stress, apoptosis, and pyroptosis of chondrocytes via targeting MYD88. Mechanistically, miR-203a-3p exerted protective effects via the inhibition of the MYD88/NF-κB pathway. In the rat OA model, intra-articular injections of miR-203a-3p agomir also significantly inhibited cartilage matrix degradation, thereby alleviating OA progression. Furthermore, the miR-203a-3p agomir-treated arthritic rat dramatically exhibited better articular tissue morphology and lower OARSI scores.
Conclusions: MiR-203a-3p plays a role in alleviating the progression of OA by regulating the MYD88/NF-κB pathway, thereby inhibiting cartilage matrix degradation, oxidative stress, apoptosis, and pyroptosis of chondrocytes. It highlights the potential significance of miR-203a-3p as an important regulator of OA.
Abbreviations
OA: osteoarthritis; miRNAs: MicroRNAs; NF-κB: nuclear factor-kappa B; IL-1β: interleukin-1β; TLR: toll-like receptor; DMEM: Dulbecco’s Modified Eagle Medium; FBS: fetal bovine serum; qRT-PCR: Quantitative real-time polymerase chain reaction; WB: Western blot; RIPA: Radio Immunoprecipitation Assay; MMP3: matrix metallopeptidase 3; MMP13: matrix metallopeptidase 13; NLRP3: NLR Family Pyrin Domain Containing 3; ASC: apoptosis-associated speck-like protein containing a CARD; GSDMD: gasdermin D; p-IκBα: phosphorylated IκBα; RT: room temperature; WT: wild type; MUT: mutant; MDA: Malondialdehyde; SOD: Superoxide Dismutase; 2,7-DCF-DA: 2,7-dichlorodihydrofluorescein diacetate; ACLT: anterior cruciate ligament transection; EDTA: ethylenediaminetetraacetic acid; OARSI: Osteoarthritis Research Society International; HE: Hematoxylin-eosin; SDs: standard deviations.