Angiotensin II Activates the Calcineurin/NFAT Signaling Pathway and Induces Cyclooxygenase-2 Expression in Rat Endometrial Stromal Cells
Figure 6
Calcium-induced COX-2 expression was not abrogated by CN inhibition in primary ESC from early pregnant rats.
ESC were isolated from uteri of pregnant rats on 4.5 day post coitus (d.p.c). (A and C) The Cox-2 mRNA was amplified from total RNA purified from primary cultures of the isolated cells by semi-quantitative RT-PCR. The transcript of β-actin 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 for 4 h to PIo (A), 500 nM of Ang II (C) (lanes 2 and 4) or left unstimulated (lanes 1 and 3). (B and D) Immunoblots showing endogenous protein expression of COX-2 and β-actin as a loading control. (B) Primary cultures of the isolated cells were pretreated for 1 h as before with vehicle (lanes 1 and 2) or CsA (lane 3), and then exposed to PIo for 8 h (lanes 2 and 3) or not (lane1). (D) Primary cultures of the isolated cells were pretreated for 1 h with vehicle (lanes 1 and 2) or CsA (lane 3 and 4), and then exposed to Ang II for 8 h (lanes 2 and 4) or not (lanes1 and 3). (A–D) Lower panels bar graphs show the densitometric data analysis of the results shown in upper panels. The COX-2/β-actin ratio was calculated and plotted against the values obtained with the control, vehicle-treated non-stimulated rat ESC (ns), which were assigned a value of 1. Open bars, cells pre-treated with vehicle; closed black bars, cells pre-treated with CsA. Values plotted are the means ± SD of the fold induction values obtained from three independent experiments. *P<0.05; ** P<0.01 (ANOVA).
