Figure 3: DHED treatment induces PR expression, an estrogen marker in the hypothalamus.

(a) DHED administration (p.o., 30 μg/kg once a day for 4 days) induces PR expression, evaluated with in situ hybridization histochemistry in the POA of the animals tested in the physiological model of hot flushes (Fig. 2b). Vehicle and EE (p.o., 200 μg/kg once a day for 4 days) were included in the experimental design as negative and positive (estrogen) controls, respectively (n = 4–6 per treatment group). (b) Optical density over the POA in DHED, EE or vehicle-treated rats. Both EE and DHED significantly increased optical density, representing PR expression in the POA compared to vehicle-treated animals. Data represent mean ± SEM. *P < 0.05 compared with vehicle-treated OVX group.