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。
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.
作者
袁力
纪建华
李敏艳
YUAN Li;JI Jian-hua;LI Min-yan(College of Clinical Medicine,Hanzhong Vocational and Technical College;Pharmaceutical College of Hanzhong Vocational and Technical College;Medical Technology College of Hanzhong Vocational and Technical College,Hanzhong 723002,China)
出处
《天然产物研究与开发》
CAS
CSCD
2023年第7期1144-1153,共10页
Natural Product Research and Development
基金
陕西省教育厅2022年度青年创新团队科研计划(22JP004)。
