Correlation between inflammatory factors in synovial cells and microRNA-145/mitogen-activated protein kinase 4 molecular axis

doi:10.3969/j.issn.2095-4344.3145

Abstract

Abstract: BACKGROUND: Inflammatory cytokines have been shown to be abnormally expressed in chondrocytes and double-knuckle cells. Targeting inflammatory pathways may be an effective strategy for the treatment of osteoarthritis, but few reports focus on inflammatory responses in synovial cells.
OBJECTIVE: To investigate the effect of 99Tc-MDP on synovial cells-produced inflammatory factors through microRNA-145 (miR-145)/mitogen-activated protein kinase 4 (MKK4) molecular axis.
METHODS: Osteoarthritis fibroblast-like synovial cells (HFLS-RA cells) were treated with 99Tc-MDP of different mass concentrations for 48 hours. The expression levels of miR-145 and inflammatory factors, caspase 1, interleukin (IL)-1, IL-18 and tumor necrosis factor-α (TNF-α) in HFLS-RA cells were measured by qRT-PCR, and the survival rate of HFLS-RA cells was measured by cell counting kit-8. After determining the optimal mass concentration for 99Tc-MDP intervention, HFLS-RA cells were divided into four groups: blank control group (without any treatment), 99Tc-MDP group (13 μg/L 99Tc-MDP treated HFLS-RA cells for 48 hours), 99Tc-MDP+miR-145 inhibitor group (13 μg/L 99Tc-MDP treated HFLS-RA cells followed by transfection with miR-145 inhibitor), 99Tc-MDP+miR-145 inhibitor+sh-MKK4 group (13 μg/L 99Tc-MDP treated HFLS-RA cells followed by transfection with miR-145 inhibitor+sh-MKK4). The expression levels of miR-145 and the inflammatory factors, caspase 1, IL-1, IL-18 and TNF-α, in HFLS-RA cells were measured by qRT-PCR, and the survival rate of HFLS-RA cell was measured by cell counting kit-8.
RESULTS AND CONCLUSION: 99Tc-MDP significantly inhibited the expression of caspase 1, IL-1, IL-18 and TNF-α as well as the cell survival rate of hFLS-RA cells. The 50% inhibiting concentration of 99Tc-MDP was 13 μg/L. 99Tc-MDP significantly upregulated the expression of miR-145, and downregulated the expression of MKK4 expression in HFLS-RA cells (P < 0.01). MiR-145 targeted MKK4, and overexpression of miR-145 could significantly inhibit the expression of MKK4. Compared with the 99Tc-MDP group, the 99Tc-MDP+miR-145 inhibitor group significantly inhibited the expression of miR-145 in HFLS-RA cells (P < 0.05) and promoted the expression of inflammatory factors in the cells (P < 0.05) and cell proliferation (P < 0.05). Compared with the 99Tc-MDP+miR-145 inhibitor+sh-MKK4 group, the 99Tc-MDP+miR-145 inhibitor group could significantly promote HFLS-RA cell proliferation (P < 0.05) and increase the expression of inflammatory factors in the cells (P < 0.05). To conclude, 99Tc-MDP relieves osteoarthritis by inhibiting HFLS-RA cell-produced inflammatory factors via the downregulation of MKK4 by miR-145.

Key words: , 99Tc-MDP, miR-145, MKK4, inflammatory factor, inflammation, synovial cells, osteoarthritis

CLC Number:

Zhu Lixuan, Cui Yue, Luo Jing. Correlation between inflammatory factors in synovial cells and microRNA-145/mitogen-activated protein kinase 4 molecular axis[J]. Chinese Journal of Tissue Engineering Research, 2021, 25(14): 2199-2204.

Figures/Tables 4

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