姜黄素及白藜芦醇对溃疡性结肠炎小鼠巨噬细胞的影响和机制  

Effect and mechanism of curcumin and resveratrol on macrophages of mice with ulcerative colitis

作　　者：张立泽[1] 王丹丹[1] 乔翠霞[1] 高海瑞[1] 尤雯丽 赵刚[1] Zhang Lize;Wang Dandan;Qiao Cuixia;Gao Hairui;You Wenli;Zhao Gang(Department of Anorectum,The Affiliated Hospital,Qingdao University,Qingdao 266000,China)

机构地区：[1]青岛大学附属医院肛肠科,266000

出　　处：《中华炎性肠病杂志（中英文）》2020年第2期131-138,共8页Chinese Journal of Inflammatory Bowel Diseases

基　　金：齐鲁卫生与健康领军人才培育工程专项经费。

摘　　要：目的探讨姜黄素和白藜芦醇对溃疡性结肠炎(UC)小鼠结肠组织巨噬细胞的影响及相关机制。方法将60只BALB/c小鼠随机平均分为CON组、DSS组、DSS+Cur组、DSS+Res组、Eve+Cur组及Eve+Res组,每组10只。CON组为正常对照组,不进行处理,其余5组均采用3%右旋葡聚糖硫酸钠(DSS)制作UC小鼠模型,即自由饮用3%DSS溶液7 d。DSS组单纯建立模型,DSS+Cur组、DSS+Res组在模型基础上分别每天给予0.4 ml姜黄素、0.4 ml白藜芦醇灌胃。Eve+Cur组在模型基础上每天给予0.4 ml姜黄素灌胃后,再给予0.4 ml mTOR抑制剂Everolimus灌胃。Eve+Res组在模型基础上每天给予0.4 ml白藜芦醇灌胃后,再给予0.4 ml mTOR抑制剂Everolimus灌胃。给药7 d后处死小鼠,分离培养各组小鼠结肠组织巨噬细胞。观察小鼠一般情况并行疾病活动指数(DAI)评分。为观察姜黄素和白藜芦醇对巨噬细胞的影响,采用CCK-8法检测巨噬细胞的增殖活性,流式细胞术检测巨噬细胞凋亡情况,荧光定量PCR和Western blot法检测巨噬细胞中自噬相关因子LC3、Beclin-1和Atg12的mRNA和蛋白表达,ELISA法检测巨噬细胞培养上清中细胞炎症因子白细胞介素-6(IL-6)、肿瘤坏死因子α(TNF-α)的水平。为进一步探讨机制,采用ELISA法检测mTOR抑制剂Everolimus对IL-6和TNF-α的影响,采用荧光定量PCR和Western blot法检测巨噬细胞SIRT1/mTOR信号通路相关因子mTOR和SIRT1 mRNA和蛋白的表达。结果小鼠无死亡。与CON组相比,DSS组、DSS+Cur组和DSS+Res组巨噬细胞的增殖活性均明显增高,凋亡率均明显降低,Atg12、Beclin-1和LC3 mRNA和蛋白的表达水平明显增加(均P<0.05);与DSS组相比,DSS+Cur组和DSS+Res组巨噬细胞的增殖活性则显著降低,凋亡率均明显增高,Atg12、Beclin-1和LC3 mRNA和蛋白的表达水平明显降低(均P<0.05)。与CON组相比,DSS组、DSS+Cur组、DSS+Res组、Eve+Cur组、Eve+Res组小鼠DAI明显升高,巨噬细胞中mTOR和SIRT1 mRNA和蛋白的表达�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-αin 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 macro

关 键 词：溃疡性结肠炎 姜黄素 白藜芦醇 巨噬细胞 自噬 

分 类 号：R28[医药卫生—中药学]