Tnf α

The supernatant of rDPSCs from each group was collected. The levels of TNF-α (ER1393, Wuhan Fine Biotech, Wuhan, China), IL-6 (MBS269892, MyBioSource, San Diego, CA, USA), IL-1β (MBS2023030, Mybiosource), and CXCL1 (ER1014, Wuhan Fine Biotech) were measured by ELISA, and the experimental procedures were strictly performed according to the kit instructions.

Birds were euthanized on days 1 and 42 of the experiment. On the first day of the experiment, 10 one-day-old chicks from each group were euthanized by decapitation to collect blood and liver tissue. The remaining 20 chickens in each group were euthanized on the 42nd D of the experiment. The livers (from all birds in each group) were immediately excised, rinsed of blood, and homogenized using a mechanical homogenizer in PBS containing 0.05% sodium azide and 0.5% Triton X-100 (pH 7.4). Liver samples were then sonicated for 10 min. All solid particles were removed by centrifugation of the homogenates at 12,000 × g for 10 min. Dichloromethane (0.4 mL) was added to 1 mL of supernatant to eliminate all lipid contaminants. This was followed by a second centrifugation for 10 min at 12,000 × g, after which the supernatant was removed and immediately assayed. ELISA kits for the determination of chicken CRP, Hp, α-1-AGP, SAA, TRF and TNF-α, IFN-γ, IL-2, and IL-10 were obtained from Wuhan Fine Biotech Co., Ltd. (East Lake High-tech Development District, Wuhan, Hubei Province, China). The ELISA experiments were performed according to the manufacturer's specifications. APP and cytokine content were expressed as ng/mL total protein.