芍药苷通过抑制TLR5表达及T细胞活化减轻葡聚糖硫酸钠诱导的小鼠溃疡性结肠炎  

Paeoniflorin attenuates dextran sulfate sodium-induced ulcerative colitis in mice by inhibiting TLR5 expression and T cell activation

作　　者：王小静 罗霞[1] 赵忠正 罗爽[1] 陈津岩[3] 周联[1] WANG Xiaojing;LUO Xia;ZHAO Zhongzheng;LUO Shuang;CHEN Jinyan;ZHOU Lian(Department of Immunopharmacology,School of Pharmaceutical Sciences,Guangzhou University of Chinese Medicine,Guangzhou 510006;Department of Critical Care Medicine,People’s Hospital of Dongping County,Taian 271000;Center for Basic Research of Chinese and Western Integration,School of Basic Medical,Guangzhou University of Chinese Medicine,Guangzhou 510006,China)

机构地区：[1]广州中医药大学中药学院免疫药理室,广东广州510006 [2]山东泰安市东平县人民医院重症医学科,山东泰安271000 [3]广州中医药大学基础医学院中西结合基础研究中心,广东广州510006

出　　处：《细胞与分子免疫学杂志》2020年第8期673-679,共7页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金(81673668);广东省自然科学基金(2017A030313657)。

摘　　要：目的探究芍药苷对葡聚糖硫酸钠(DSS)诱导的溃疡性结肠炎小鼠的治疗作用及其机制。方法将C57BL/6雄性小鼠随机分为正常组、模型组、600 mg/(kg·d)美沙拉嗪处理组、(12.5、25、50)mg/(kg·d)芍药苷处理组,每组10只。除正常组外,其他组给予30 g/L DSS溶液造模5 d,同时连续灌胃给药10 d,正常组与模型组给予等量的蒸馏水。每天观察小鼠体质量、粪便性状和便血情况,评分并计算疾病活动指数(DAI);HE染色观察结肠组织变化,ELISA检测血清鞭毛蛋白(flagellin)抗体、白细胞介素6(IL-6)和肿瘤坏死因子α(TNF-α)含量;Western blot法检测小鼠结肠组织Toll样受体5(TLR5)、髓样分化因子88(MyD88)、核因子κBp65(NF-κBp65)蛋白水平;流式细胞术检测肠系膜淋巴结的淋巴细胞活化情况。结果与正常组相比,模型组小鼠DAI评分显著升高,结肠长度明显缩短;黏膜上皮、肠腺等结构消失;血清中鞭毛蛋白抗体、IL-6和TNF-α含量升高;结肠组织TLR5、MyD88、NF-κBp65蛋白表达明显增加;肠系膜淋巴结T淋巴细胞活化增加。与模型组比较,美沙拉嗪与各剂量芍药苷组的上述症状均得到缓解。结论芍药苷通过抑制TLR5的表达以及T细胞活化,减轻小鼠溃疡性结肠炎。Objective To investigate the therapeutic effect and mechanism of paeoniflorin on dextran sulphate sodium(DSS)-induced ulcerative colitis(UC)mice.Methods C57BL/6 male mice were randomly divided into control group,model group,600 mg/(kg·d)mesalazine treatment group,(12.5,25,50)mg/(kg·d)paeoniflorin treatment group,with 10 mice in each.All mice were treated with 30 g/L DSS for 5 days except the control group.Meanwhile,the mice in the other groups were orally administrated corresponding drugs for 10 days,while the mice in the control and model groups were given equivalent volumes of distilled water.Body mass,fecal characteristics and hematochezia of the mice were observed and recorded daily,and then disease activity index(DAI)was evaluated and calculated.Pathological changes in the colon were observed by HE staining.The levels of anti-flagellin antibody,interleukin-6(IL-6)and tumor necrosis factor alpha(TNF-α)in the serum were measured by ELISA.The expression levels of Toll-like receptor 5(TLR5),myeloid differentiation factor 88(MyD88)and nuclear factor kappa-Bp65(NF-κBp65)in the colon tissues were evaluated by Western blot analysis and the activation of lymphocytes in mesenteric lymph node(MLN)was detected by flow cytometry.Results Compared with the control group,DAI scores in the model group were significantly raised,the colon length was significantly shortened,and the epithelium and intestinal gland disappeared.In addition,the serum levels of anti-flagellin antibody,IL-6,TNF-αand the protein levels of TLR5,MyD88,NF-κBp65 in the colon significantly increased,and the activation of T lymphocytes in MLN went up in the model group.All symptoms above were alleviated in the mesalazine and paeoniflorin groups compared with the model group.Conclusion Paeoniflorin can attenuate UC in mice by inhibiting the expression of flagellin and TLR5,and the activation of T cells.

关 键 词：芍药苷 溃疡性结肠炎 Toll样受体5(TLR5) 核因子κBp65(NF-κBp65) T细胞活化 

分 类 号：R285.5[医药卫生—中药学] R574.62[医药卫生—中医学]