Abstract: Objective To explore the effect and mechanism of acupuncture at Zusanli (ST36) on inflammatory response in mice with mesenteric lymphadenitis based on the mitogen-activated protein kinase (MAPK)/nuclear factor-κB (NF-κB) signaling pathway. Methods Twelve male Kunming strain mice were randomly selected as blank control group, and mice with successfully established mesenteric lymphadenitis models were randomly divided into model group and acupuncture at Zusanli group, with 12 mice in each group. Mice in the acupuncture at Zusanli group received electroacupuncture treatment at Zusanli for 4 days, while those in the blank control group and the model group received no treatment. Behavioral characteristics of mice in each group were observed. Hematoxylin and eosin (HE) staining was used to evaluate pathological changes in mesenteric lymph node tissues of mice. Enzyme-linked immunosorbent assay (ELISA) was employed to detect the expression levels of pro-inflammatory cytokinestumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), interleukin-6 (IL-6)in mouse serum. Western blot was used to detect the expression of autophagy-related proteins (Beclin1, LC3-Ⅱ, LC3-Ⅰ, p62) and MAPK/NF-κB signaling pathway-related proteins (p38 MAPK, p-p38 MAPK, NF-κB p65, p-NF-κB p65) in mesenteric lymph node tissues of mice. Results Mice in the model group exhibited decreased food intake, lethargy, soft and loose stools, and occasional kyphosis. The behavioral characteristics of mice in the acupuncture at Zusanli group were improved compared with those in the model group. HE staining results showed pathological changes in mesenteric lymph node tissues of mice in the model group, while the pathological changes in the acupuncture at Zusanli group were improved compared with those in the model group. ELISA results showed that the levels of TNF-α, IL-1β, and IL-6 in serum of mice in the model group were higher than those in the blank control group, while those in the acupuncture at Zusanli group were lower than those in the model group (P<0.05). Western blot results showed that the relative expression of Beclin1 protein and the LC3-Ⅱ/LC3-Ⅰ in mesenteric lymph node tissues of mice in the model group were lower than those in the blank control group, while the relative expression of p62 protein and the ratios of p-p38 MAPK/p38 MAPK and p-NF-κB p65/NF-κB p65 were higher than those in the blank control group (P<0.05). In the acupuncture at Zusanli group, the relative expression of Beclin1 protein and the LC3-Ⅱ/LC3-Ⅰ were higher than those in the model group, while the relative expression of p62 protein and p-p38 MAPK/p38 MAPK and p-NF-κB p65/NF-κB p65 were lower than those in the model group (P<0.05). Conclusion Acupuncture at Zusanli alleviates the inflammatory response in mice with mesenteric lymphadenitis by inhibiting the p38 MAPK/NF-κB signaling pathway.