Fig. 4. Activation of mAChRs down-regulated neutrophils migration and reduced proinflammatory cytokines level. (A) Neutrophils were photographed with a fluorescence microscope (n = 25), and neutrophils migration was reduced after treatment with bethanechol (1 μM, 10 μM) in the statistical area. The caudal fin was not injured in the right picture, and the caudal fin was injured in the left picture. (B) Model of caudal fin injury: The bethanechol (1 μM, 10 μM) treated or untreated caudal fin was cut with a scalpel blade (white rectangles indicate statistical areas). (C) When the zebrafish caudal fin was not damaged, there was no difference in the number of neutrophils in the statistical area between bethanechol (1 μM, 10 μM) and the control group (the white rectangles indicate the statistical area). (D) The expression levels of tnf-α, il-1β, il-6 and il-8 in zebrafish treated with bethanechol (1 μM, 10 μM) were decreased compared with the LPS group (n = 30). (*P < 0.05，** P < 0.01, ***P < 0.001, ****P < 0.0001, one-way ANOVA analysis, LSD for port-hoc test).
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