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Updated: Jan 4, 2026
Software-Assisted Quantitative Measurement of Osteoarthritic Subchondral Bone Thickness
Published on: March 18, 2022
MIR-140-5p affects chondrocyte proliferation, apoptosis, and inflammation by targeting HMGB1 in osteoarthritis
Yingjie Wang1, Songpo Shen2, Zeng Li1
1Department of Orthopedics, Peking Union Medical College Hospital, Peking Union Medical College and Chinese Academy of Medical Science, No.1 Shuaifuyuan, Dongcheng District, Beijing, 100730, China.
Objective:
This study aimed to test the expression and biological function of miR-140-5p in osteoarthritis (OA), and identify its target gene and explore its mechanism in OA.
Methods:
Differential genes were screened and analyzed by gene microarray and WGCNA analysis. The normal human chondrocytes C28/I2 were induced by IL-1β to construct the OA cell model. The expression of miR-140-5p and high mobility group box 1 (HMGB1) was quantified by quantitative real-time PCR (qRT-PCR) in OA tissues and IL-1β-induced chondrocytes. Western blotting was performed to evaluate the expression of HMGB1 and PI3K/AKT pathway activation. The concentrations of tumor necrosis factor (TNF)-α, interleukin (IL)-6, MMP-1 and MMP-3 were determined by ELISA. CCK-8 and flow cytometry were conducted to determine the cellular capabilities of proliferation and cell apoptosis.
Results:
Bioinformatics analysis demonstrated that HMGB1 was highly expressed in OA and activated PI3K/AKT pathway. Also, HMGB1 was predicted as a target of miR-140-5p. The levels of miR-140-5p were negatively correlated with HMGB1 in OA tissues and IL-1β-induced chondrocytes. The overexpression of miR-140-5p reduced the expression of HMGB1 protein, p-AKT (Ser473) and p-PI3K in IL-1β-induced chondrocytes. Besides, the expression of p-AKT (Ser473) and p-PI3K was significantly upregulated by employing miR-140-5p inhibitor, but retrieved after treating with LY294002. Furthermore, miR-140-5p inhibited inflammation, matrix metalloprotease expression and apoptosis in IL-1β-induced chondrocytes through regulating HMGB1.
Conclusion:
MiR-140-5p was down-regulated while HMGB1 was upregulated in OA. MiR-140-5p could inhibit the PI3K/AKT signaling pathway and suppress the progression of OA through targeting HMGB1.
Insights
Osteoarthritis (OA) shows decreased miR-140-5p and increased high mobility group box 1 (HMGB1). MiR-140-5p suppresses OA progression by inhibiting the PI3K/AKT pathway via targeting HMGB1.
Area of Science:
- Molecular Biology
- Cell Biology
- Biochemistry
Background:
- Osteoarthritis (OA) is a degenerative joint disease characterized by cartilage breakdown.
- MicroRNAs (miRNAs) play crucial roles in regulating gene expression and are implicated in OA pathogenesis.
- Dysregulation of specific miRNAs, such as miR-140-5p, may contribute to OA development.
Purpose of the Study:
- To investigate the expression and function of miR-140-5p in osteoarthritis.
- To identify high mobility group box 1 (HMGB1) as a target gene of miR-140-5p.
- To elucidate the mechanism by which miR-140-5p influences OA progression via the PI3K/AKT signaling pathway.
Main Methods:
- Gene microarray and WGCNA analysis to identify differential genes in OA.
- Construction of an OA cell model using IL-1β-induced human chondrocytes (C28/I2).
- Quantification of miR-140-5p and HMGB1 expression using qRT-PCR; assessment of PI3K/AKT pathway activation via Western blotting; measurement of inflammatory cytokines and matrix metalloproteinases using ELISA; evaluation of cell proliferation and apoptosis via CCK-8 and flow cytometry.
Main Results:
- HMGB1 was upregulated in OA tissues and IL-1β-induced chondrocytes, correlating negatively with miR-140-5p levels.
- Overexpression of miR-140-5p reduced HMGB1 expression and inhibited the PI3K/AKT pathway.
- MiR-140-5p suppressed inflammation, matrix metalloproteinase production, and apoptosis in OA chondrocytes by regulating HMGB1.
Conclusions:
- MiR-140-5p is downregulated, while HMGB1 is upregulated in osteoarthritis.
- MiR-140-5p exerts protective effects in OA by targeting HMGB1 and inhibiting the PI3K/AKT signaling pathway.
- Targeting the miR-140-5p/HMGB1 axis represents a potential therapeutic strategy for osteoarthritis.
