大黄灵仙方调控胆管细胞炎症的作用及机制研究  

Mechanism of Dahuang Lingxian Formula in alleviating LPS-induced inflammation in rat bile duct cells

作　　者：陈伟棠 俞渊[1] 庞浇安 李敏鹏 潘孟[1] 陆世锋[1] 杨文 滕金豪 叶桂源 CHEN Weitang;YU Yuan;PANG Jiaoan;LI Minpeng;PAN Meng;LU Shifeng;YANG Wen;TENG Jinhao;YE Guiyuan(The First Affiliated Hospital of Guangxi University of Chinese Medicine,Nanning 530023,China)

机构地区：[1]广西中医药大学第一附属医院肝胆外科,南宁530023

出　　处：《环球中医药》2024年第2期209-216,共8页Global Traditional Chinese Medicine

基　　金：广西自然科学基金(2020GXNSFAA238012);2021“岐黄工程”高层次人才团队培育项目(2021006);2020年广西中医药大学第一附属医院院级博士启动基金(2020BS004);2020年广西中医药大学引进博士科研启动基金(2020BS030)。

摘　　要：目的 观察大黄灵仙方调控白细胞介素-1受体相关激酶(interleukin-1 receptor-associated kinase 1,IRAK)-1和IRAK-4以及成纤维生长因子-3激活酶1MAP3K7结合蛋白2(TGF-3 activator 1 map3k7-binding protein 2,TAB2)的表达水平,探讨其缓解胆管细胞炎症的作用机制,为预防肝胆管结石的形成及术后复发提供理论依据。方法 将45只SPF级健康雄性SD大鼠随机分为9组,空白组、模型组、大黄灵仙颗粒组、激活核转录因子κB(nuclear factor kappa-B,NF-κB)阻断剂组、p38丝裂活化蛋白激酶(mitogen-activated protein kinase, MAPK)阻断剂组、NF-κB阻断剂+p38MAPK阻断剂组、NF-κB阻断剂+大黄灵仙颗粒组、p38MAPK阻断剂+大黄灵仙颗粒组、NF-κB阻断剂+p38MAPK阻断剂+大黄灵仙颗粒组,每组大鼠5只。除空白组以外,其余各组大鼠于胆总管处一次性注射5 mg/kg内毒素脂多糖以构建肝内胆管感染动物模型。第7天灌胃结束后取材,取大鼠完整胆管树组织,采用蛋白免疫印迹法和实时荧光定量PCR检测IRAK-1、IRAK-4、TAB2蛋白及mRNA表达量。结果 (1)与空白组比较,模型组IRAK-1、IRAK-4、TAB2蛋白及mRNA表达量升高(P<0.05);(2)与模型组比较,大黄灵仙颗粒组、NF-κB阻断剂组、p38MAPK阻断剂组、NF-κB阻断剂+p38MAPK阻断剂组、NF-κB阻断剂+大黄灵仙颗粒组、IRAK-1、IRAK-4、TAB2的蛋白及mRNA表达量降低(P<0.05),NF-κB阻断剂+p38MAPK阻断剂+大黄灵仙颗粒组IRAK-1、IRAK-4、TAB2蛋白及TAB2 mRNA表达量均显著下降(P<0.05);(3)与大黄灵仙颗粒组比较,NF-κB阻断剂+大黄灵仙颗粒组、NF-κB阻断剂+p38MAPK阻断剂+大黄灵仙颗粒组IRAK-1、IRAK-4、TAB2的蛋白表达量和NF-κB阻断剂+p38MAPK阻断剂组的IRAK-1、IRAK-4、TAB2 mRNA表达量均下降(P<0.05)。结论 大黄灵仙方可缓解内毒素脂多糖诱导的大鼠胆管炎症反应,抑制炎症因子的异常表达,促使胆道恢复至非炎症状态,从而达到预防肝胆管结石的形成及术后复发Objective To observe the expression levels of IRAK-1 and IRAK-4 and TAB2 regulated by Dahuang Lingxian formula,and to explore its mechanism in alleviating bile duct cell inflammation,so as to provide theoretical basis for preventing the formation of hepatobiliary stones and postoperative recurrence.Methods Forty-five SPF-grade healthy male SD rats were randomly divided into nine groups,blank group,model group,Dahuang Lingxian Formula group,NF-κB blocker group,p38MAPK blocker group,NF-κB blocker+p38MAPK blocker group,NF-κB blocker+Dahuang Lingxian Formula group,p38MAPK blocker+Dahuang Lingxian Formula group,NF-κB blocker+p38MAPK blocker group,NF-κB blocker+p38MAPK blocker+Dahuang Lingxian Formula group,5 rats in each group.Except for the blank group,the rats in each group were injected with 5 mg/kg LPS at the common bile duct at one time to construct the animal model of intrahepatic bile duct infection.After gavage on the 7th day,the intact bile duct tree tissues were taken from the rats,and IRAK-1,IRAK-4,TAB2 protein and mRNA expression were detected by Western blot and RT-qPCR.Results(1)Compared with the blank group,the protein and mRNA expressions of IRAK-1,IRAK-4,and TAB2 were increased in the model group(P<0.05);(2)compared with the model group,the protein and mRNA expressions of IRAK-1,IRAK-4,and TAB2 were increased in the Dahuang Lingxian Formula group,NF-κB blocker group,p38MAPK blocker group,NF-κB blocker+p38MAPK blocker group,NF-κB blocker+Dahuang Lingxian Formula group.The expression levels of IRAK-1,IRAK-4、TAB2 protein and TAB2 mRNA in the NF-κB blocker+p38MAPK blocker+Dahuang Lingxian Formula group were reduced(P<0.05);(3)compared with the Dahuang Lingxian Formula group,the NF-κB blocker+Dahuang Lingxian Formula group,NF-κB blocker+p38MAPK blocker+Dahuang Lingxian Formula group IRAK-1,IRAK-4,TAB2 protein and mRNA expression and NF-κB blocker+p38MAPK blocker group,IRAK-1,IRAK-4 and TAB2 mRNA expression levels are reduced(P<0.05).Conclusion Dahuang Lingxian Formula can relieve the inflamma

关 键 词：大黄灵仙方 胆管炎症 肝内胆管结石 白细胞介素-1受体相关激酶1 白细胞介素-1受体相关激酶4 成纤维生长因子-3激活酶1MAP3KT结合蛋白2 

分 类 号：R244.1[医药卫生—针灸推拿学]