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Influence of Anemoside B4 on ulcerative colitis in mice based on regulation of macrophage polarization |
SUN Jie1 SHI Xiaohua1 LI Caiyun2 JI Shiliang2 |
1.Department of Gastroenterology, Suzhou Science & Technology Town Hospital, Jiangsu Province, Suzhou 215153, China;
2.Department of Pharmacy, Suzhou Science & Technology Town Hospital, Jiangsu Province, Suzhou 215153, China
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Abstract Objective To investigate the effect of Anemoside B4 on ulcerative colitis (UC) induced by dextran sulfate sodium (DSS) in mice based on regulation of macrophage polarization. Methods Forty SPF male C57BL/6 mice (body weight 18-20 g) were divided into control group, model group, B4 group, and positive drug group by random number table method, with ten mice in each group. Model group, B4 group, and positive drug group were given 3%DSS for five days consecutively to construct UC model. After successful modeling, B4 group was given intragastric administration of Anemoside B4 (5 mg/kg, once a day), positive drug group was given intragastric administration of Mesalazine (400 mg/kg, once a day), control group and model group were given intragastric administration of the same amount of normal saline. All four groups were given continuous intervention for seven days. The changes of mice body weight during the intervention were observed. After the intervention, the colon tissues of mice were stained with hematoxylin-eosin, and the histopathological index was further evaluated, and the inflammatory indicators of four groups were detected. The mRNA expression levels of nuclear factor-κB (NF-κB) p50, p65, and M1 macrophages type specific index inducible nitric oxide synthase (iNOS), M2 macrophages type specific index arginine-1 (Arg1) in colon tissues of the four groups were detected by qRT-PCR. The polarization of macrophages in the colon tissues of the four groups was observed by immunohistochemical staining. Results The body weight of the model group was lower than those of the control group at 6, 9, and 12 d after intervention, while the body weight of the B4 group at 12 d after intervention was higher than that of the model group (P<0.01 or P<0.05). In the control group, mucosal intestinal glands were arranged neatly without ulcerative lesions. In the model group, most of the crypts and epithelium of the intestinal mucosal layer were lost, accompanied by a large number of inflammatory cell infiltration, inflammatory invasion of the mucosal layer and submucosa. In B4 group, the colonic mucosa had slight inflammation, ulcer, and bleeding symptoms were significantly reduced, the epithelium and recess were relatively intact, and the colonic mucosa injury was also significantly improved. Histopathological index, levels of tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, IL-6, and mRNA expression levels of NF-κB p50, NF-κBp65, iNOS in model group were higher than those in control group, levels of IL-10, transforming growth factor-β (TGF-β), and mRNA expression level of arginine-1 (Arg1) were lower than those in control group (P<0.05). Histopathological index, levels of TNF-α, IL-1β, IL-6, and mRNA expression levels of NF-κBp50, NF-κBp65, iNOS in B4 group were lower than those in model group, levels of IL-10, TGF-β, and mRNA expression level of Arg1 were higher than those in model group (P<0.05). The proportion of M1 macrophages in the model group was higher than that in the control group, and the proportion of M2 macrophages was lower than that in the control group, the proportion of M1 macrophages in the B4 group was higher than that in the model group, and the proportion of M2 macrophages was higher than that in the model group (P<0.05). Conclusion Anemoside B4 can inhibit the M1 macrophages, promote the M1 macrophages, improve the intestinal inflammatory environment and relieve UC.
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