2.6. Animals and Experimental Design

In preparation for drug administration, eriodictyol was accurately weighed and dissolved in PBS to obtain concentrations of 1, 2, and 4 mg/mL. Silibinin was dissolved in PBS to achieve a concentration of 10 mg/mL. The dosage and administration method for eriodictyol were determined based on previous investigations [10,15].

Sixty male ICR mice (20 ± 2 g), aged 4 weeks, were obtained from Chengdu Dassy Biotechnology Co., Ltd. The mice were housed in a controlled environment with a constant temperature of (23 ± 2 °C), relative humidity of (55 ± 5%), and a 12-h light-dark cycle. They had ad libitum access to food and water. After a 7-day acclimatization period, the mice were randomly divided into six experimental groups as follows: (1): The control group (n = 10) received an intraperitoneal injection of physiological saline. (2) The LPS/D-GalN group (ALI model, n = 10) received an intraperitoneal injection of physiological saline. (3) The LPS/D-GalN+eriodictyol low-dose treatment group (eriodictyol 10 mg/kg, n = 10) received an intraperitoneal injection of eriodictyol at a dose of 10 mg/kg. (4) The LPS/D-GalN+eriodictyol medium-dose treatment group (eriodictyol 20 mg/kg, n = 10) received an intraperitoneal injection of eriodictyol at a dose of 20 mg/kg. (5) The LPS/D-GalN+eriodictyol high-dose treatment group (eriodictyol 40 mg/kg, n = 10) received an intraperitoneal injection of eriodictyol at a dose of 40 mg/kg. (6) The LPS/D-GalN+silibinin treatment group (silibinin 100 mg/kg, n = 10) received an intraperitoneal injection of silibinin at a dose of 100 mg/kg.

The administration was carried out once a day for a duration of 7 days. On the 7th day, mice in the LPS/D-GalN, LPS/D-GallN+eriodictyol (10, 20, and 40 mg/kg), and LPS/D-GalN+silibinin groups were intraperitoneally injected with 600 mg/kg of D-GalN and 10 μg/kg of LPS, following one hour after the administration of eriodictyol, silibinin, or physiological saline. The control group received a saline injection alone. After 6 h, all mice were euthanized to collect blood and tissue samples. Plasma was collected, followed by centrifugation at 3500× g for 10 min at 4 °C to obtain the supernatant, which was then stored at −80 °C.