4.2. Carrageenan-induced inflammatory air pouch mouse model

Male ICR mice (20–25 g) were purchased from Orient Bio Inc. (Seongnam, Korea). In order to make space (or air pouch) under the skin, 2 mL of air was injected thrice into the intra-scapular area of the mice’s back for six days. The animals were subsequently divided into two categories of which one (n = 6) was not treated with carrageenan while the other (n = 24) was treated with carrageenan; the animal study was undertaken twice using the same protocol. The carrageenan-treated category consisted of four groups including an inflammation-induced group that had been injected with carrageenan after normal saline oral administration, a 5 mg/kg indomethacin-treated group (used as an anti-inflammatory drug), a 30 mg/kg A. hookeri-treated group, and a 300 mg/kg A. hookeri-treated group. Indomethacin and A. hookeri were orally administered. At two hours after treatment, the carrageenan solution (1 mL, 2 w/v % dissolved in saline) was injected into the air pouch and after 24 h of carrageenan injection, all mice were sacrificed using with over-dosing of Zoletil (Virbac, Carros, France) via intraperitoneal injection. To collect the exudate, the pouches were flushed with 2 mL of phosphate buffered saline (PBS). The number of total and differential cells in the exudate from the pouch was counted using the Hemavet Multispecies Hematology System (Drew Scientific Inc., Waterbury, CT, USA).