Downregulation of iNOS, IL-1β, and P2X7 Expression in Mast Cells via Activation of PAR4 Contributes to the Inhibition of Visceral Hyperalgesia in Rats

Figure 5

Effect of MAPK on PAR4-AP-evoked tryptase, iNOS, IL-1β, and P2X7 protein and mRNA expressions in BMMCs. (a) Western blotting analyses for tryptase, iNOS, IL-1β, and P2X7 protein expression in BMMCs following treatment with PAR4-AP and pretreatment with U-46619 (an activator of ERK1/2 and p38), tBHQ (an activator of ERK1/2), PD98059 (an inhibitor of ERK1/2), or SB203580 (an inhibitor of p38). The mean optic densities of the proteins were calculated by normalizing to GADPH. (b) Quantitative RT-PCR analyses of tryptase, iNOS, IL-1β, and P2X7 mRNA expression in BMMCs following treatment of the BMMCs with PAR4-AP and pretreatment with U-46619, tBHQ, PD98059, or SB203580. The results were calculated by normalizing to β-actin in the same sample with the ΔCt method. The changes in the relative mRNA levels are expressed as fold changes compared with the controls. All values are expressed as the means ± SEMs (). versus controls; versus PAR4-AP-only groups.

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