Balb c mice

Six-week-old male BALB/c mice were purchased from Daehan Biolink (Choongchung, Korea) and maintained in a room controlled at 23 ± 2 °C with a relative humidity of 50–55% and a 12/12 h light/dark cycle. To induce colitis, the DSS-treated mice were fed 5% DSS dissolved in sterile distilled water for 7 days. The control mice received plain drinking water only. CA (30 or 300 mg/kg body weight) and a positive drug control, SAL (100 mg/kg body weight) dissolved in olive oil, were administered concomitantly by gavage once a day for 7 days. The mice in the control group and colitis group were given only the vehicle (olive oil, 0.2 mL/day). Water consumption did not differ between groups in the experiment. This study was approved by the Institutional Animal Care and Use Committee of Korea Food Research Institute (#KFRI-M-110006), and animal care was in accordance with institutional guidelines.

Seven-week-old male BALB/c mice were purchased from Daehan Biolink (DBL; Seoul, Korea) and housed in the laboratory animal facility at Pusan National University. They were provided food and water ad libitum. The mice were housed in standard plastic cages (two mice per cage) with sawdust as bedding and maintained under controlled conditions of temperature at 22-24°C, humidity at 60 ± 5%, and alternating light/dark cycles (lights were on between 7:00 h and 19:00 h). There were likewise provided with standard laboratory food and water. The animal protocol used in this study was reviewed and approved by the Pusan National University–Institutional Animal Care Committee (PNU–IACUC) in accordance to procedure ethicality and scientific care.

Female five-week-old BALB/c mice were purchased from Daehan Biolink (Seoul, Korea). They were housed in the laboratory animal facility at Kyung Hee University (Seoul, Korea) and provided ad libitum water and food. The Kyung Hee University Institutional Animal Care Committee reviewed and approved the protocol with respect to ethical issues and scientific care (Approval Number, KHSASP-20-197).

A specific pathogen free male BALB/c mice aged 8 weeks were purchased from Daehan
Biolink (Eumseong, Korea) and experimented under aseptic management (breeding
room temperature, 25°C; humidity, 55%; and sterile distilled water
supply). Mice were divided into groups administered 250 and 500 mg/kg of WP and
HWPH, respectively, and a control group that was administered isotonic sodium
chloride solution. Eight mice were included in each group. Gastric intubation
was performed five times per week for 4 weeks. Body weight was measured twice a
week at 3-day intervals. After administration, blood was collected and
centrifuged at 2,000×g for 10 min to separate the serum.
After sacrifice, aseptically excised splenocytes for cell culture were dispensed
into 24 well-culture plates and cultured in a CO₂ incubator for 48 h.
Subsequently, phytohemagglutinin (Sigma-Aldrich, 5
μg/L×10⁶ cells) was added as a polyclonal
stimulator for in vitro activation. The concentration of
cytokines (INF-γ and IL4), antibodies (IgG2a and IgG1) and IgE in the
cell culture medium were measured using an ELISA kit purchased from BD
Bioscience (San Diego, CA, USA). For the small intestine cytokines, 2 mL of PBS
was added to 0.5 g of duodenum aseptically extracted after sacrifice, vortexed,
and left at 4°C for 2 days. Thereafter, the supernatant obtained by
centrifugation was analyzed for cytokines and IgE using the ELISA kits.