Figure 7
Attenuation of colitis by AQ in vivo. (A–C) C57BL6 mice were administered with normal water (no DSS, n = 6) or 3% DSS water (DSS, n = 12) for 6 days. DSS-treated mice were injected with either vehicle (DSS + veh, n = 6) or AQ (10 mg/kg, DSS + AQ, n = 6) every day. Three independent experiments were performed for statistical analysis. Body weight (n = 15 per group) was measured and weight loss was calculated compared to that on day 0. **p < 0.005 (A). Colon length was determined and expressed as mean ± SD of 6 mice. **p < 0.005 and ***p < 0.0005 (B). Representative images of HE and PAS staining of colon sections (n = 6) (C). (D–G) Naïve CD4+ T cells were isolated from C57BL6 mice and adoptively transferred to RAG KO mice (RAG KO/CD4+ T, n = 12). After 4 weeks, mice were injected daily with vehicle (n = 6) or AQ (10 mg/kg, n = 6) for 2 weeks. This experiment was repeated twice and DAI (n = 12 per group) was determined by observing the weight loss, diarrhea, and rectal bleeding. ***p < 0.0005 (D). Colon tissue sections (n = 6 per group) were stained with HE or fluorescence-tagged antibodies against CD4 and Foxp3. DPI stained nuclei and representative images are shown (E). Total proteins (n = 3) were harvested from spleen and subjected to immunoblotting analysis of FoxP3, RORγt, T-bet, and β-actin. Representative blots of three independent experiments are shown (F). Colon tissues (n = 6) were excised and cultured in DMEM for 24 h, and cell supernatants were used for determining IL-17 and IFN-γ by ELISA. *p < 0.05 and ***p < 0.0005 (G).
