Unlike for Human Monocytes after LPS Activation, Release of TNF-α by THP-1 Cells Is Produced by a TACE Catalytically Different from Constitutive TACE
Figure 7
Effects of Ro 32-7315, Celastrol and TQ on MOCA and ABZ substrates hydrolysis by THP-1 cells.
The viable intact THP-1 cells (0.5×106 cells/ml) were incubated with 5 µM of the MOCA or ABZ substrate in Ca2+ and Mg2+ free HBSS in the presence of various concentrations of Ro 32-7315, Celastrol or TQ. The final DMSO concentration in the reaction mixture was 0.1%. The fluorescence intensity was monitored for 600 s and the rates of the substrates hydrolysis were calculated from the linear section of the fluorescence curve. To estimate the accurate inhibitory potency of the tested compounds, the reaction rates of MOCA hydrolysis obtained for different compounds concentrations (V(MOCA)[inhibitor]) were divided by MOCA hydrolysis reaction rate in the presence of 0.1%DMSO (V(MOCA)[0.1% DMSO]). Inhibitory potency (%) = 100%−(V(MOCA)[inhibitor]/V(MOCA)[0.1% DMSO])×100%.