Angiotensin II Activates the Calcineurin/NFAT Signaling Pathway and Induces Cyclooxygenase-2 Expression in Rat Endometrial Stromal Cells
Figure 2
Ang II induced COX-2 expression in primary ESC of non-pregnant rats. Expression was abrogated by CN inhibition.
(A) Cox-2 mRNA was amplified from total RNA purified from primary cultures of ESC, by semi-quantitative RT-PCR. The transcript of the β-actin gene was used as internal control. Cells were pretreated for 1 h with vehicle (lanes 1 and 2), or 200 ng/ml of CsA (lanes 3 and 4), and then exposed to 500 nM of Ang II for 4 h (lanes 2 and 4) or left unexposed (lanes 1 and 3). (B) Immunoblots of whole extracts of primary cultured ESC isolated from uteri of non-pregnant rats showing endogenous protein expression of COX-2 and β-actin as a loading control. Primary cultures of the cells were pretreated as before for 1 h with vehicle (lanes 1 and 2), or CsA (lanes 3 and 4), and then exposed to 500 nM Ang II (lanes 2 and 4), or left untreated (lanes 1 and 3) for 8 h. (A and B) Right panel 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 performed. Open bars, cells pre-treated with vehicle; closed black bars, cells pre-treated with CsA *** P<0.001; ** P<0.01 (ANOVA).
