Icariin

Herba Epimedii, Radix Achyranthis Bidentatae, Radix Astragali, and Concha Ostreae were purchased from Anhui Xiehecheng Co., Ltd. (Bozhou, China) and evaluated by Doctor Rongchun Han (College of Pharmacy, Anhui University of Chinese Medicine, Hefei, China). The standard substances of calycosin (Lot number: MUST-18120901), calycosin-7-glucoside (Lot number: MUST-19031920), ononin (Lot number: MUST-19041101), epimedin C (Lot number: MUST-19081310), icariin (Lot number: MUST-18091010), and ecdysterone (Lot number: MUST-18120209) were purchased from Chengdu Must Bio-Technology Co., Ltd. (Chengdu, China). Epimedin A (Lot number: JOT-10354), epimedin B (Lot number: JOT-10353), and ginsenoside Ro (Lot number: JOT-10448) were purchased from Chengdu Pufei De Biotech Co., Ltd. (Chengdu, China). The purity of all standards was over 98.0%. Acetonitrile (HPLC grade; Lot number: SHBM1775) and methanol were purchased from Sigma Aldrich Trading Co., Ltd. (Shanghai, China). Formic acid (HPLC grade; Lot number: J2027129) was obtained from Aladdin Biochemical Technology Co., Ltd. (Shanghai, China). Ultrapure water was prepared by a Milli Q-Plus system (Millipore, Bedford, USA).

Chengdu Must Bio-Technology Co., Ltd. provided the compositions of the ECC-BYF III, which are icariin (MUST-17051810), astragaloside IV (MUST-17022804), nobiletin (MUST-16070901), 20-S-ginsenoside Rh1 (MUST-17030717), and paeonol (MUST-16071405). Aminophylline tablets were purchased from Xinhua Pharmaceutical Co., Ltd. (Shandong, China). Both ECC-BYF III and aminophylline were fully dissolved in 0.5% CMC-Na (sodium carboxymethyl cellulose; Hengxing Chemical Reagent Manufacturing Co., Ltd.; 500 g/bottle) and prepared into suspension before gavage. Acupuncture needles were purchased from Suzhou Acupuncture Supplies Co., Ltd. (Huanqiu Brand), and its specification is 0.30 mm × 13 mm, 100 pieces per box. The electroacupuncture apparatus was purchased from Suzhou Medical Supplies Factory Co., Ltd.