Chinese Journal of Inflammatory Bowel Diseases 2020;04(2):131-138
doi:10.3760/cma.j.cn101480-20190606-00083
Effect and mechanism of curcumin and resveratrol on macrophages of mice with ulcerative colitis
Lize ZHANG 1 ; Dandan WANG 1 ; Cuixia QIAO 1 ; Hairui GAO 1 ; Wenli YOU 1 ; Gang ZHAO 1
Affiliations
Keywords
Ulcerative colitis; Curcumin; Resveratrol; Macrophages; Autophagy
Country
China
Language
Chinese
Abstract
Objective:To explore the effects and related mechanisms of curcumin and resveratrol on macrophages of colon tissues in ulcerative colitis (UC) mice.Methods:Sixty BALB/c mice were randomly and equally divided into CON group, DSS group, DSS+Cur group, DSS+Res group, Eve+Cur group and Eve+Res group. There were 10 mice in each group. CON group was the normal control group and received no treatment. UC model of mice was established by giving 3% dextran sodium sulfate (DSS) to drink freely for 7 days in other 5 groups. UC model of mice was simply established in DSS group. The gavage administrations of 0.4 ml curcumin and 0.4 ml resveratrol were performed every day in the mice of DSS+Cur group and DSS+Res group respectively after establishing the UC model. The gavage administration of 0.4 ml mTOR inhibitor Everolimus was performed in Eve+Cur group after 0.4 ml curcumin gavage administration and also was performed in Eve+Res group after 0.4 ml resveratrol gavage administration every day. The mice were sacrificed after 7 days of administration. The macrophages of colon tissues were isolated and cultured in each group. The general conditions of the mice were observed and the DAI score was evaluated. In order to examine the effects of curcumin and resveratrol on macrophages, CCK-8 method was used to detect the proliferation and flow cytometry was used to detect the apoptosis of macrophages. Fluorescence quantitative PCR and Western blot were used to detect the mRNA and protein expressions of the autophagy-related factors LC3, Beclin-1 and Atg12. ELISA was used to detect the level of the inflammatory factors IL-6 and TNF-α in culture supernatants of macrophages. In order to explore the associated mechanism, ELISA was used to detect the influence of mTOR inhibitor Everolimus on IL-6 and TNF-α, fluorescence quantitative PCR and Western blot were used to detect the expressions of SIRT1/mTOR pathway-related factors mTOR and SIRT1 mRNA and protein.Results:No death was observed in all of the mice. Compared with CON group, the proliferative activity of macrophages was significantly higher, and the apoptosis rate decreased significantly, the mRNA and protein expressions of Atg12, Beclin-1 and LC3 increased significantly in DSS Group, DSS+Cur group and DSS+Res group (all P<0.05) . Compared with DSS group, the proliferative activity was significantly lower, the apoptosis rate increased obviously, the mRNA and protein expressions of Atg12, Beclin-1 and LC3 decreased significantly in DSS+Cur group and DSS+Res group (all P<0.05) . Compared with CON group, the DAI scores increased, the mRNA and protein expressions of mTOR and SIRT1 decreased significantly, the levels of IL-6 and TNF-α increased significantly in DSS group, DSS+Cur group, DSS+Res group, Eve+Cur group and Eve+Res group (all P<0.05) . Compared with DSS group, the DAI scores decreased, the mRNA and protein expressions of mTOR and SIRT1 increased significantly, the levels of IL-6 and TNF-α decreased significantly in DSS+Cur group and DSS+Res group (all P<0.05) . Compared with DSS+Cur group and DSS+Res Group, the DAI scores increased significantly, the mRNA and protein expressions of mTOR and SIRT1 decreased significantly, the levels of IL-6 and TNF-α increased significantly in Eve+Cur group and Eve+Res group (all P<0.05) . Conclusion:Curcumin and resveratrol can inhibit the macrophages proliferation of colon tissues in UC mice, promote the apoptosis and inhibit the development of inflammation and autophagy, which may be regulated by SIRT1/mTOR pathway.
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