Donepezil hydrochloride

SLF was prepared at a concentration of 100 mg/mL at the School of Pharmacy, Southwest Medical University (Luzhou, Sichuan, China) using a 1.0 g suspension equivalent to 31.5 g crude material. After purchasing donepezil hydrochloride (lot: 110823A) from Eisai Inc. (Suzhou, Jiangsu, China), a 0.1 mg/mL concentration was prepared. Aβ_25-35 (lot: Y-0044) was acquired from BIOSS Biotech Co., Ltd. (Beijing, China). Anti-AKT and anti-GSK-3β antibodies were purchased from Abcam (Cambridge, MA, USA). Anti-tau antibody was obtained from American Bioworld Co., Ltd. (Dublin, OH, USA). Anti-GAPDH antibody was purchased from Kangchen Bio-tech Inc. (Shanghai, China). The brain stereotaxic apparatus was purchased from Huaibei Zhenhua Biological Equipment Co., Ltd. (Anhui, Huaibei, China). The Morris water maze was purchased from Chengdu Taimeng Technology Co., Ltd. (Chengdu, Sichuan, China). An S70 Inverted microscope was purchased from Leica DMIRB (Rueil-Malmaison, France). The ChemiDocXRS gel imaging and analysis system was purchased from Bio-Rad Laboratories, Inc. (Berkeley, CA, USA). SP-9000 immunohistochemistry kit was purchased from Beijing ZSGB Biological Engineering Co., Ltd. (Beijing, China). ECL Western detection reagents were from 4A Biotech Co., Ltd. (Beijing, China).

Donepezil hydrochloride (Eisai China Inc.) was directly dissolved in sterile distilled water. CDS extract, encompassed danshen, sanqi and bingpian in proportions of 450:141:8, was provided by Guangzhou Baiyunshan Hutchison Whampoa Chinese Medicine Co.Ltd. The compound danshen was prepared as previous study [10 (link)]. Briefly, Salvia miltiorrhiza (450 g), Notoginseng (141 g) and Borneolum Syntheticum (8 g), were extracted with 4.8 L ethanol heating reflux for 1.5 h, filtered, ethanol were recycled and filtered solution were concentrated to moderate volume; then the dregs were extracted with 4.8 L 50 % ethanol heating reflux for 1.5 h, filtered, ethanol were recycled and filtrate were concentrated to moderate volume; dregs were extracted with 3.6 L water for 2 h, filtered, filtrate were concentrated to moderate volume; Radix Notoginseng were crushed into fine powder, combined with the above concentrate solution and the moderate auxiliary materials made into grain, dried; Borneolum Syntheticum were porphyrized, blending with the grain. It was dissolved in sterile distilled water. And both mixtures were stored at 4 °C for use.
Aβ_25–35 (Sigma-Aldrich, St. Louis, Missouri) was dissolved in sterile distilled water at a concentration of 1 mg/ml as a stocking solution. Aβ_25–35 was “aged” by incubation at 37 °C for 4 days and stored at −20 °C for use [16 (link), 17 (link)].