Effects of DON on the expression of inflammatory cytokines, CD44, MMP-13, and integrin in cultured chondrocytes / 中华地方病学杂志

ABSTRACT

Objective:To observe the effects of deoxynivalenol (DON) on the expression of chondrocyte inflammatory cytokines, cluster of differentiation (CD) 44, matrix metalloproteinase (MMP)-13 and integrin in vitro. Methods:Primary chondrocytes from Wistar rats aged 1 to 2 days were isolated, and the chondrocytes were identified by toluidine blue staining. The effects of DON (dose groups of 0.0, 0.1, 0.2, 0.4, 0.8 μg/ml) on proliferation of chondrocytes were detected by CCK-8 method for 24, 48 and 72 h. According to the cell survival rate, the DON concentrations [0.00 (control), 0.05, 0.10, 0.15, 0.20 μg/ml] were selected for subsequent experiments, and the exposure time was 48 h. The contents of interleukin (IL)-6, IL-1β, tumor necrosis factors (TNF)-α, CD44 and MMP-13 in the cell culture supernatant were detected by enzyme linked immunosorbent assay (ELISA) kit. The expression levels of CD44 and integrin subunits α2, α5, and β1 protein in chondrocytes were detected by Western blotting.Results:The survival rate of chondrocytes decreased with the increase of DON concentrations and time ( P < 0.01). There were statistically significant differences in IL-6 content between different groups ( H = 13.425, P < 0.01), and 0.10 μg/ml group [256.89 (191.02, 477.58) pg/ml] was significantly higher than that of the control group [10.37 (0.00, 119.13) pg/ml, P < 0.05]. There were no statistically significant differences in IL-1β and TNF-α contents between the groups ( F = 0.881, 1.317, P > 0.05). The CD44 contents in 0.10, 0.15, and 0.20 μg/ml groups [(0.87 ± 0.21), (0.85 ± 0.24), (0.77 ± 0.17) pg/ml] were lower than that in the control group [(1.06 ± 0.19) pg/ml, P < 0.05]. There were statistically significant differences in MMP-13 expression between the groups ( F = 7.947, P < 0.01). There were statistically significant differences in protein expression of CD44, integrin subunits α2, α5 and β1 between the groups ( F = 5.737, 6.562, 6.074, 4.476, P < 0.05 or < 0.01). Conclusion:DON may disrupt the balance between cells and extracellular matrix by increasing the expression of IL-6, MMP-13, integrin subunits α2 and β1, and inhibiting the expression of CD44 and integrin subunit α5, thus causing cartilage injury.