Angiotensin II Activates the Calcineurin/NFAT Signaling Pathway and Induces Cyclooxygenase-2 Expression in Rat Endometrial Stromal Cells
Figure 1
Calcium-induced expression of COX-2 was dependent on calcineurin activity in primary ESC of non-pregnant rats.
Primary ESC were isolated from uteri of non-pregnant rats and cultured during 96 h. (A) Cox-2 mRNA was amplified from total RNA by semi-quantitative RT-PCR. The transcript of β-actin was used as internal control. Primary cells were pretreated for 1 h with vehicle (lanes 1–2), 200 ng/ml of CsA (lanes 3–4), or 10 µg/ml of actinomycin D (AcD, lanes 5–6), and then exposed to PIo (a combination of 20 ng/ml of PMA plus 1 µM calcium ionophore Io, for 4 h (lanes 2, 4, 6), or left untreated (lanes 1, 3, 5). (B) Immunoblots of whole extracts obtained from ESC showing endogenous protein expression of COX-2 and β-actin as a loading control. Cells were pretreated as before for 1 h with vehicle (lanes 1–2) or CsA (lanes 3–4) and then exposed to PIo, for 8 h (lanes 2–4). (A and B). Right panels bar plots show the densitometric data analysis of the results shown in the left panels A and B. The COX-2/β-actin ratio was calculated and plotted against the values obtained with the control, non-stimulated rat ESC, which were assigned a value of 1 (ns). The values plotted are the means ± SD of the fold induction values obtained from three independent experiments. Open bars, cells pre-treated with vehicle; closed black bars, cells pre-treated with CsA; closed gray bars, cells pre-treated with AcD. *** P<0.001; ** P<0.01 (ANOVA).
