Dipirona50

At 3 months of age, the rats were ovariectomized. Surgery was performed through an abdominal ventral incision under deep anesthesia with sodium pentobarbital (60 mg/kg, i.p., Cheminova de México, México City, México; Reg. SAGARPA Q-7048-044) and atropine sulfate (0.05 mg/kg, i.p., Sigma-Aldrich, St. Louis, MO, USA). The oviducts and ovaries were ligated and subsequently removed. The tissue area was carefully cleaned with benzalkonium chloride (Medipharm®, San Luis Río Colorado, Sonora, México), and then the muscle and skin were sutured separately. Analgesic-antipyretic medication (50 mg/kg Dipirona50®, i.m., Virbac Animal Health, Guadalajara, México) was administered to minimize postsurgical pain for 4 days after surgery. After surgery, the rats were returned to the housing facilities for 12 weeks to ensure the long-term absence of ovarian hormones and confirm an increase in anxiety-like behavior [19 (link), 23 (link)]. Afterward, the rats were randomly assigned to the experimental groups, received their respective treatments, and underwent behavioral testing.

Pentobarbital sodium was purchased from Cheminova de Mexico, Mexico City, Mexico. Reg. SAGARPA Q-7048-044). Benzalkonium chloride from Medipharm^®, San Luis Río Colorado, Sonora, Mexico. Dipirona50^® was obtained from Virbac Animal Health, Guadalajara, Mexico. Dimethylsulfoxide (DMSO) was purchased from Golden Bell Reactivos (Mexico City, Mexico). Atropine sulphate, chrysin (Chry, 5,7-Dihydroxyflavone < 97%) and allopregnanolone (Allo, 5α-Pregnan-3α-ol-20-one) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Diazepam (Valium, injectable solution) was obtained from Laboratory Roche (Mexico City, Mexico). Saline solution (0.9%) was purchased from PiSA Farmacéutica (Guadalajara, Jalisco, Mexico).