天然产物研究与开发 ›› 2023, Vol. 35 ›› Issue (7): 1144-1153.doi: 10.16333/j.1001-6880.2023.7.005

• 研究论文 • 上一篇    下一篇

木犀草素调节SIRT3/AMPK/mTOR信号通路对溃疡性结肠炎小鼠Th17/Treg免疫平衡的影响

袁   力1*,纪建华2,李敏艳3   

  1. 1汉中职业技术学院临床医学院;2汉中职业技术学院药学院;3汉中职业技术学院医学技术学院,汉中 723002
  • 出版日期:2023-07-31 发布日期:2023-07-31
  • 基金资助:
    陕西省教育厅2022年度青年创新团队科研计划(22JP004)

Effect of luteolin on Th17/Treg immune balance in mice with ulcerative colitis by regulating SIRT3/AMPK/mTOR signaling pathway

YUAN Li1*,JI Jian-hua2,LI Min-yan3   

  1. 1College of Clinical Medicine,Hanzhong Vocational and Technical College;2Pharmaceutical College of Hanzhong Vocational and Technical College;3Medical Technology College of Hanzhong Vocational and Technical College,Hanzhong 723002,China
  • Online:2023-07-31 Published:2023-07-31

摘要:

探究木犀草素对溃疡性结肠炎(ulcerative colitis,UC)小鼠模型中辅助性T细胞17(Th17)/调节性T细胞(Treg)免疫平衡的影响,并分析其潜在机制。60只BALB/c雄性小鼠随机分为正常组、模型组、阳性对照组、木犀草素组、木犀草素+3-TYP组,每组12只。采用葡聚糖硫酸钠(DSS)诱导建立UC模型,观察并记录小鼠体重变化、大便稠度和大便潜血情况,计算疾病活动指数(DAI);HE染色观察结肠组织病理学变化;流式细胞术检测脾脏Th17、Treg细胞比例,计算Th17/Treg比值;ELISA检测结肠组织中IL-6、IL-10、IL-17和IL-23含量;RT-qPCR检测结肠组织RORγt和Foxp3的mRNA表达;Western blot检测结肠组织SIRT3、AMPK、p-AMPK、mTOR、p-mTOR蛋白表达。与正常组相比,模型组小鼠DAI评分、组织病理学评分、脾脏Th17/Treg比值、结肠组织IL-6、IL-17、IL-23、RORγt mRNA水平和p-mTOR/mTOR比值升高,IL-10、Foxp3 mRNA和SIRT3蛋白水平以及p-AMPK/AMPK比值降低(P<0.05)。经药物干预后,小鼠DAI评分、组织病理学评分、脾脏Th17/Treg比值、结肠组织IL-6、IL-17、IL-23、RORγt mRNA水平和p-mTOR/mTOR比值降低,IL-10、Foxp3 mRNA和SIRT3蛋白水平以及p-AMPK/AMPK比值升高(P<0.05);3-TYP可减弱木犀草素对UC小鼠Th17/Treg细胞分化平衡的影响(P<0.05)。木犀草素可改善DSS诱导的UC小鼠模型中Th17/Treg失衡,其机制可能与激活SIRT3/AMPK/mTOR通路有关。

关键词: 溃疡性结肠炎, 木犀草素, 辅助性T细胞17, 调节性T细胞, 沉默信息调节因子3

Abstract:

To explore the effect of luteolin on the immune balance of T helper 17 cells (Th17)/regulatory T cells (Treg) in a mouse model of ulcerative colitis (UC),and to analyze its underlying mechanism.Sixty BALB/c male mice were randomly separated into normal group,model group,positive control group,luteolin group,luteolin+3-TYP group,with 12 mice in each group.Dextran sodium sulfate (DSS) was performed to induce the establishment of a UC model,the weight change,stool consistency and stool occult blood in mice were observed and recorded,and the disease activity index (DAI) was calculated;HE staining was used to observe the histopathological changes of the colon;flow cytometry was used to measure the ratio of Th17 and Treg cells in the spleen,and the ratio of Th17/Treg was calculated;ELISA was used to measure the contents of IL-6,IL-10,IL-17 and IL-23 in colon tissue;RT-qPCR was used to measure the mRNA expression of RORγt and Foxp3 in colon tissue;Western blot was used to measure the expression of colonic tissue SIRT3,AMPK,p-AMPK,mTOR,p-mTOR proteins.Compared with the normal group,the DAI score,histopathology score,spleen Th17/Treg ratio,colon tissue IL-6,IL-17,IL-23,RORγt mRNA levels and p-mTOR/mTOR ratio of mice increased in the model group,the IL-10,Foxp3 mRNA and SIRT3 protein levels,and p-AMPK/AMPK ratio decreased (P<0.05).After drug intervention,the DAI score,histopathology score,spleen Th17/Treg ratio,colon tissue IL-6,IL-17,IL-23,RORγt mRNA levels and p-mTOR/mTOR ratio of mice decreased,the IL-10,Foxp3 mRNA and SIRT3 protein levels,and p-AMPK/AMPK ratio increased P<0.05);3-TYP was able to weaken the effect of luteolin on the differentiation balance of Th17/Treg cells in UC mice (P<0.05).Luteolin can improve Th17/Treg imbalance in DSS-induced UC mouse model,and its mechanism may be related to the activation of SIRT3/AMPK/mTOR pathway.

Key words: ulcerative colitis, luteolin, T helper 17 cells, regulatory T cells, silent information regulator 3

中图分类号:  R285.5 TQ041