The Effects of Amphiregulin Induced MMP-13 Production in Human Osteoarthritis Synovial Fibroblast
Figure 1
Concentration- and time-dependent increases in MMP-13 production by AREG. ((a) and (b)) Human synovial fibroblasts were obtained from normal or osteoarthritis patients and examined by Western blot and qPCR for the expression of AREG. (c) OASF and normal SF were incubated with AREG for 24 h. The mRNA expression of MMPs was examined by qPCR . ((d) and (e)) OASF and normal SF were incubated with various concentrations of AREG for 24 h or with AREG (50 ng/mL) for 6, 12, or 24 h. The mRNA expression of MMP-13 was examined by qPCR . ((f) and (g)) OASFs were incubated with various concentrations of AREG for 24 h or with AREG (50 ng/mL) for 6, 12, or 24 h, and supernatants and cell lysates were then collected. MMP-13 protein levels in cell lysates were determined by Western blot analysis. MMP-13 enzymatic activity in cell lysates and supernatants was measured using zymography. Both protein levels and enzymatic activity increased in a time-dependent manner . (h) The protein and mRNA levels of AREG and MMP-13 in control-shRNA and AREG-shRNA OASF were examined by Western blot and qPCR. Results are expressed as the mean ± SEM. compared with control; compared with AREG-treated group.
