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Published on: October 4, 2024
MiR-29a and MiR-140 Protect Chondrocytes against the Anti-Proliferation and Cell Matrix Signaling Changes by IL-1β
Xianghui Li1, Zhilei Zhen2, Guodong Tang2
1Department of Pharmacy, the Second Affiliated Hospital of Harbin Medical University, Harbin, China.
Abstract:
As a degenerative joint disease, osteoarthritis (OA) constitutes a major cause of disability that seriously affects the quality of life of a large population of people worldwide. However, effective treatment that can successfully reverse OA progression is lacking until now. The present study aimed to determine whether two small non-coding RNAs miR-29a and miR-140, which are significantly down-regulated in OA, can be applied together as potential therapeutic targets for OA treatment. MiRNA synergy score was used to screen the miRNA pairs that potentially synergistically regulate OA. An in vitro model of OA was established by treating murine chondrocytes with IL-1β. Transfection of miR-29a and miR-140 via plasmids was investigated on chondrocyte proliferation and expression of nine genes such as ADAMTS4, ADAMTS5, ACAN, COL2A1, COL10A1, MMP1, MMP3, MMP13 and TIMP metal-lopeptidase inhibitor 1 (TIMP1). Western blotting was used to determine the protein expression level of MMP13 and TIMP1, and ELISA was used to detect the content of type II collagen. Combined use of miR-29a and miR-140 successfully reversed the destructive effect of IL-1β on chondrocyte proliferation, and notably affected the MMP13 and TIMP1 gene expression that regulates extracellular matrix. Although co-transfection of miR-29a and miR-140 did not show a synergistic effect on MMP13 protein expression and type II collagen release, but both of them can significantly suppress the protein abundance of MMP13 and restore the type II collagen release in IL-1β treated chondrocytes. Compared with single miRNA transfection, cotransfection of both miRNAs exceedingly abrogated the suppressed the protein production of TIMP1 caused by IL-1β, thereby suggesting potent synergistic action. These results provided novel insights into the important function of miRNAs' collaboration in OA pathological development. The reduced MMP13, and enhanced TIMP1 protein production and type II collagen release also implies that miR-29a and miR-140 combination treatment may be a possible treatment for OA.
Insights
This study shows that combining miR-29a and miR-140 can reverse osteoarthritis damage in cells. This miRNA combination may offer a new treatment for osteoarthritis by reducing MMP13 and increasing TIMP1 and collagen II.
Area of Science:
- Biochemistry
- Molecular Biology
- Genetics
Background:
- Osteoarthritis (OA) is a degenerative joint disease causing significant disability worldwide.
- Current treatments for OA lack the ability to reverse disease progression.
- MicroRNAs (miRNAs) miR-29a and miR-140 are downregulated in OA and are investigated as potential therapeutic targets.
Purpose of the Study:
- To evaluate the therapeutic potential of co-administering miR-29a and miR-140 for osteoarthritis treatment.
- To investigate the synergistic effects of miR-29a and miR-140 on chondrocyte function and extracellular matrix gene expression.
- To explore the role of miRNA collaboration in OA pathogenesis.
Main Methods:
- An in vitro OA model using IL-1β-treated murine chondrocytes.
- Screening for miRNA pairs with synergistic regulatory potential using miRNA synergy score.
- Transfection of miR-29a and miR-140 via plasmids.
- Analysis of chondrocyte proliferation, gene expression (ADAMTS4, ADAMTS5, ACAN, COL2A1, COL10A1, MMP1, MMP3, MMP13, TIMP1), protein levels (MMP13, TIMP1) via Western blotting, and type II collagen release via ELISA.
Main Results:
- Combined miR-29a and miR-140 reversed IL-1β-induced chondrocyte proliferation defects.
- Co-transfection significantly impacted MMP13 and TIMP1 gene expression.
- While not synergistic for MMP13 protein or collagen II release, both miRNAs suppressed MMP13 and restored collagen II.
- Co-transfection showed synergistic action by abrogating IL-1β-induced TIMP1 suppression.
Conclusions:
- miR-29a and miR-140 exhibit collaborative function in OA pathological development.
- Combined treatment with miR-29a and miR-140 reduced MMP13, enhanced TIMP1 production, and restored type II collagen release.
- This miRNA combination represents a potential therapeutic strategy for osteoarthritis.
