Donepezil d6821

BIS-MEP was synthesized by the Xie lab in the Department of Chemistry, School of Pharmacy, Fudan University. IBO (I2765) and donepezil (D6821) were purchased from Sigma-Aldrich (Atlanta, GA, USA). The Aβ monoclonal antibody, 6E10, was obtained from Covance (Emeryville, CA, USA). Glial fibrillary acidic protein (GFAP) monoclonal antibody was purchased from Millipore (Temecula, CA, USA). The polyclonal antibody against ionized calcium-binding adapter molecule 1 (IBA1), a microglia-specific protein, was obtained from Arigo Biolaboratories (Taipei, Taiwan, China). Aβ_1–42 (03112), Amplex Red Acetylcholine/AChE Assay Kit (A12217) and the Pierce BCA Protein Assay Kit (23225) were purchased from Thermo Fisher Scientific (Rockford, IL, USA). Mouse TNF-α and mouse IL-6 ELISA kits were obtained from ExCell Biology (Shanghai, China). All other reagents were purchased from commercial sources.

Four-week-old ICR male mice weighing approximately 25-30 g (Orient Co., Seongnam, Korea) were first placed in a breeding room under the following conditions: temperature of 22 ± 2°C, 50 ± 5% humidity, and 12-h dark/12-h light cycle. The mice were adapted to the environment for one week with water and feed supplied ad libitum. The animals were then divided into four groups with six animals per group: the control group, which was administered saline; the scopolamine group, which was administered scopolamine ((−)-scopolamine hydrobromide trihydrate, S1875, Sigma-Aldrich, St. Louis, MO, USA) intraperitoneally at a dose of 1 mg/kg to induce memory impairment; the donepezil group, which was administered 6 mg/kg donepezil (D6821, Sigma-Aldrich, St. Louis, MO, USA); and the positive control group, which was administered 1 mg/kg and 5 mg/kg β-carotene (C4582, Sigma-Aldrich, St. Louis, MO, USA) [17 , 18 ]. The fermented S. maxima extract was also administered at doses of 50, 100, 200, and 400 mg/kg with saline water.