芍药甘草汤减轻炎性肠病大鼠炎症反应和肠黏膜屏障损伤  

Shaoyao-Gancao decoction attenuates inflammatory response and intestinal mucosal barrier injury in rats with inflammatory bowel disease

作　　者：刘重洋 吴苗苗[2] 王越 冯志芳 杨越雯 蔺双双 贺伟[1] LIU Chongyang;WU Miaomiao;WANG Yue;FENG Zhifang;YANG Yuewen;LINShuangshuang;HE Wei(College of Traditional Chinese Medicine,Hebei North University,Zhangjiakou 075000,China;College of Pharmacy,Hebei North University,Zhangjiakou 075000,China)

机构地区：[1]河北北方学院中医学院,河北张家口075000 [2]河北北方学院药学院,河北张家口075000

出　　处：《中国病理生理杂志》2024年第9期1678-1687,共10页Chinese Journal of Pathophysiology

基　　金：河北省高等学校科学技术研究青年基金项目(No.QN2022014);河北北方学院校级课题项目(No.XJ2021022);河北省大学生创新创业项目(No.S202310092023)。

摘　　要：目的:探讨芍药甘草汤对2,4,6-三硝基苯磺酸(TNBS)诱导的炎性肠病(inflammatory bowel disease,IBD)大鼠炎症反应及肠黏膜屏障损伤的作用。方法:48只雄性SD大鼠随机分为:正常组,模型组,高剂量(500 mg/kg)、中剂量(250 mg/kg)和低剂量(125 mg/kg)芍药甘草汤组,巴柳氮钠(1 g/kg)组。所有组别预给药3 d,实验第4天禁食不禁水24 h,除正常组外,其余5组大鼠均以TNBS(100 mg/kg)与50%乙醇等体积混匀进行灌肠造模,造模结束后继续给药5 d,并对大鼠进行疾病活动度指数(disease activity index,DAI)评估,实验结束后,测定大鼠血清及结肠组织中一氧化氮(nitric oxide,NO)和髓过氧化物酶(myeloperoxidase,MPO)的水平,RT-qPCR及Western blot法测定大鼠结肠组织中肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)、环加氧酶2(cyclooxygenase-2,COX-2)、诱导型一氧化氮合酶(inducible nitric oxide synthase,iNOS)和核因子κB(nuclear factor-κB,NF-κB)的表达水平,免疫荧光法测定大鼠结肠组织中紧密连接蛋白[闭锁小带蛋白1(zonula occludens-1,ZO-1)和密封蛋白2(claudin 2)]的表达。结果:与正常组相比,模型组大鼠体重下降、结肠缩短,DAI及结肠组织宏观评分显著升高(P<0.05),结肠病理损伤严重,模型组大鼠血清及结肠组织中NO和MPO水平显著升高(P<0.05),结肠组织中TNF-α、COX-2、iNOS和NF-κB的mRNA及蛋白表达水平显著升高(P<0.01),紧密连接蛋白ZO-1和claudin 2的表达水平显著降低(P<0.01)。芍药甘草汤干预能有效缓解上述状况,与模型组相比,芍药甘草汤组大鼠体重下降和结肠缩短有所缓解,结肠组织DAI及结肠组织宏观评分显著降低(P<0.05),结肠病理损伤有所减轻,大鼠血清及结肠组织中NO和MPO水平显著降低(P<0.05),结肠组织中TNF-α、COX-2、iNOS和NF-κB的mRNA及蛋白表达水平显著降低(P<0.05),ZO-1和claudin 2的表达显著增加(P<0.05)。结论:芍药甘草汤能够有效减轻TNBS诱导大鼠IBD所引发的炎症反�AIM:To investigate the effects of Shaoyao-Gancao decoction(SGD)on inflammation and mucosal barrier damage in rats with 2,4,6-trinitrobenzenesulfonic acid(TNBS)-induced inflammatory bowel disease(IBD).METHODS:Forty-eight male SD rats were randomly divided into normal group,model group,high-dose(500 mg/kg),medium-dose(250 mg/kg)and low-dose(125 mg/kg)SGD groups,and balsalazide sodium(1 g/kg)group.All rats were pre-administered for 3 d,and on the 4th day of the experiment,they were fasted for 24 h.Except for the normal group,the rats in the other 5 groups were given enema mixed with TNBS(100 mg/kg)and 50%ethanol,and continued to be administered for 5 d after modeling.After modeling,the disease activity index(DAI)was evaluated.After the experiment,the levels of nitric oxide(NO)and myeloperoxidase(MPO)in serum and colonic tissues of rats were determined.RT-qPCR and Western blot were used to determine tumor necrosis factor-α(TNF-α),cyclooxygenase-2(COX-2),inducible nitric oxide synthase(iNOS)and nuclear factor-κB(NF-κB)in the colon of rats.The expression of tight junction proteins zonula occludens-1(ZO-1)and claudin 2 in rat colon tissues was determined by immunofluorescence staining.RESULTS:Compared with normal group,the weight of rats in model group was decreased,the colon was shortened,DAI and colon tissue macroscopic scores were significantly increased(P<0.05),colon pathological injury was serious,and NO and MPO levels in serum and colon tissues of the rats in model group were significantly increased(P<0.05).The mRNA and protein expression levels of TNF-α,COX-2,iNOS and NF-κB in colon tissues were significantly increased(P<0.01),while the expression levels of ZO-1 and claudin 2 were significantly decreased(P<0.01).Compared with model group,the body weight and colon shortening of rats in SGD groups were alleviated,DAI and macroscopic scores of colon tissues were significantly decreased(P<0.05),the pathological damage of colon was improved,and the levels of NO and MPO in serum and colon tissues of rats were sig

关 键 词：芍药甘草汤 炎性肠病 炎症 肠黏膜屏障 

分 类 号：R574[医药卫生—消化系统] R363.2[医药卫生—内科学] R285.5[医药卫生—临床医学]