Thrombin Induces COX-2 and PGE₂ Expression via PAR1/PKCalpha/MAPK-Dependent NF-kappaB Activation in Human Tracheal Smooth Muscle Cells

Figure 4

Thrombin induces COX-2 expression via MAPKs. (a) Cells were pretreated with SB202190, U0126, or SP600125 for 1 h and then incubated with thrombin for 6 h. The levels of COX-2 protein were determined by Western blot. (b) Cells were pretreated with U0126 (10 μM), SB202190 (10 μM), or SP600125 (10 μM) for 1 h and then incubated with thrombin for 4 h. The mRNA levels and promoter activity of COX-2 were determined. (c) Cells were transfected with scrambled, p38, p42, or JNK2 siRNA and then incubated with thrombin for 6 h. The levels of p38, p42, JNK2, and COX-2 protein were determined. (d) Cells were pretreated without or with SB202190 (10 μM), SP600125 (10 μM), U0126 (10 μM), or Gö6976 (3 μM) for 1 h and then incubated with thrombin for the indicated time intervals. The levels of phospho-p38 MAPK, phospho-p42/p44 MAPK, and phospho-JNK1/2 were determined by Western blot. (e) The media saved from (a) were used to determine the levels of PGE2 generation. Data are expressed as of three independent experiments. ^#, as compared with the control or pretreatment with inhibitor indicated in the figure.

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