Buchi b 480

The CP samples were purchased from Onggi Korean Herbal Medicine Market (Daegu, Korea) figurein April 2019, and herbal identification was performed by Professor Young-Cheol Lee, College of Korean Medicine, Sangji University, Wonju, Korea. A voucher specimen (no. 2019-SJCP-1) was deposited in the laboratory of the Department of Herbology, College of Korean Medicine, Sangji University Wonju 26339, Republic of Korea. Dried and chopped CP (600 g) was extracted three times with 70% ethanol using a 3 h reflux. The materials were filtered under reduced pressure at 40 °C using a vacuum rotatory evaporator (BUCHI B-480, Buchi, Flawil, Switzerland) and dried in a freeze-drier (EYELA FDU-540, Japan) to yield the CP extract (29.56 g). The yield (w/w) of the extract was approximately 4.93%. Dexamethasone (Sigma Aldrich, Korea), which was dissolved in saline, was used as a positive control. Methacholine, OVA, OA, and aluminum hydroxide were purchased from Sigma-Aldrich. All other chemicals and solvents used in the experiments were of analytical grade and were purchased from Sigma-Aldrich, unless otherwise indicated.

Citrus unshiu peel was supplied from Bonchowon (Yeongcheon-si, Gyeongsangnam-do) and used. 10 times water was added to 300 g of dried Citrus unshiu peel from herbs and extracted at 100°C for 2 hours using a hot water extractor (Daewoong Bio, DWT-1800T, Hwaseong, Korea). After filtration, the extraction solvent was evaporated at 50°C using a rotary evaporator (Buchi B-480, Buchi Labortechnik AG, Flawil, Switzerland). Then, the solvent was evaporated in vacuo using a freeze dryer (LABCONCO, Kansas, MO, USA) to obtain the extract in a yield of 9.3%, and Citrus unshiu peel water extract (CUP) powder was stored at −80°C.