Acetylcholinesterase

Foxtail millet husk (dry) was acquired from Jinsu Agricultural Technology Corporation (Henan Province, China). The sample was smashed (RS-FS1401 mill, Royalstar Co., Ltd., Hefei, China) sieved (40-mesh), and then stored at −20 °C.
Betaine and L-lactic acid were obtained from Shanghai Macklin Biochemical Co., Ltd., (Shanghai, China). Acetylcholinesterase, α-glucosidase (from Saccharomyces cerevisiae), and standards (vanillic acid, p-hydroxybenzoic acid, p-hydroxybenzaldehyde, caffeic acid, p-coumaric acid, ferulic acid, syringic acid, rutin, and kaempferol, the purity above 98%) were sourced from Shanghai Yuanye Biotechnology Co., Ltd., (Shanghai, China), while ABTS(2,2-azinobis-(3-ethylbenzothiazoline-6-sulfonate)), DPPH(2,2′-diphenyl-1-picrylhydrazyl), Folin–Ciocâlteu reagent, Trolox (6-hydroxy-2,5,7,8-tetramethylchromane-2-carboxylic acid), and TPTZ(2,4,6-tris(2-pyridyl)-s-triazine) were acquired from Sigma-Aldrich Chemical Co. (St. Louis, MO, USA). Chromatographic methanol and formic acid were purchased from Thermo Fisher Scientific Reagent Co., Ltd., (Waltham, MA, USA). All other reagents, including glycerol, sodium carbonate (Na₂CO₃), sodium acetate, sodium hydroxide (NaOH), sodium nitrite (NaNO₂), glycol, aluminum trichloride (AlCl₃), ferric chloride (FeCl₃), and methanol, were purchased from Tianjin Deen Chemical Reagent Co., Ltd. (Tianjing, China).

Rutin (purity > 99.0%), 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2′-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), and 5,5′-dithiobis-(2-nitrobenzoic acid) (DTNB) were purchased from Aladdin Reagent Int. (Shanghai, China). Acetylcholinesterase was purchased from Yuanye Bio-Technology Co. (Shanghai, China). Acetylthiocholine (ATCh) was purchased from Macklin Bioc-Technology Co. (Shanghai, China).

Butyrlycholinesterase (BuChE), acetylcholinesterase (AChE, from electron eel), acetylcholine iodide (AChEI), butyrylcholine iodide (BuChEI), and 5,5-dithio-bis-nitrobenzoic acid (DTNB) were purchased from Shanghai Yuanye Bio-Technology Co., Ltd. (Shanghai, China). Mouse ACh kit and AChE kit were offered by Yi Fei Xue Biotechnology Co., Ltd. (Nanjing, China). Mouse SOD kit and MDA kit were obtained from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). BDNF and TrkB antibodies were purchased from Nanjing Jin Yibai Biological Technology Co., Ltd. (Nanjing, China). 2,2-Diphenyl-1-picrylhydrazyl (DPPH) was obtained from TCI Chemical Industry Co., Ltd. (Shanghai, China); 2,2′-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) was purchased from Sigma-Aldrich (USA). Ginkgo ketoester tablets were offered by Jiangsu Shenlong Pharmaceutical Co., Ltd. (Yancheng, China) with at least 30% contents of flavonoids and lactones; donepezil (10 mg/piece) was purchased from Eisai China Inc.; scopolamine hydrobromide was obtained from Aladdin Reagents (Shanghai, China); and ultra-pure water was purified by an EPED super purification system (Nanjing, China). Other reagents and chemicals were of analytical grade.