四白汤对乙醇诱导下小鼠胃溃疡的防治作用研究  

Research on the protective effect of Sibai Decoction on ethanol-induced gastric ulcers in mice

作　　者：李平香 高瑞阳 李景尧 郑艳[2] LI Pingxiang;GAO Ruiyang;LI Jingyao;ZHENG Yan(School of Integrative Medicine,Qingdao University Medical Department,Qingdao 266023,China;Qingdao Traditional Chinese Medicine Hospital(Hiser Medical of Qingdao),Affiliate of Qingdao University Qingdao Municipal Haici Hospital,Qingdao 266033,China)

机构地区：[1]青岛大学医学部中西医结合学科,青岛266023 [2]青岛市中医医院(市海慈医院)青岛大学附属青岛市海慈医院,青岛266033

出　　处：《天津中医药》2024年第8期1030-1038,共9页Tianjin Journal of Traditional Chinese Medicine

基　　金：青岛市2015-2016年度中医药研究计划(2015-zyy013)。

摘　　要：[目的]探讨四白汤方(SBT)对乙醇(EtOH)诱导下胃溃疡(GU)模型小鼠胃黏膜损伤的防治作用及其分子机制,为其临床治疗GU提供理论依据。[方法]体外实验将5组小鼠灌服高、中、低剂量SBT 7 d后取血分离血清,用于培养细胞建立血清学研究模型。含药血清的培养基培养人胃黏膜上皮细胞-1(GES-1),细胞进入G0期后加入0.8 mol/L EtOH处理4 h建立GES-1细胞损伤模型,采用细胞计数试剂盒-8(CCK8)法检测EtOH处理后的细胞的活力和SBT含药血清对细胞增殖的影响。体内实验将60只小鼠平均分为6组,SBT预处理后,EtOH诱导小鼠胃黏膜损伤模型。观察SBT对小鼠体质量和进食量的影响,苏木精-伊红(HE)染色检测各组胃黏膜病理学损伤状况,酶联免疫吸附测定(ELISA)法检测血清中炎性标志物炎症相关白细胞介素6(IL-6)、白细胞介素1β(IL-1β)、肿瘤坏死因子α(TNF-α)和白细胞介素10(IL-10)蛋白的表达水平,生化检测试剂盒检测胃黏膜丙二醛(MDA)、活性氧簇(ROS)、还原型谷胱甘肽过氧化物酶(GSH-Px)和一氧化氮(NO)的水平,蛋白质免疫印迹分析(Western blot)检测磷脂酰肌醇3激酶(PI3K)/蛋白激酶B(AKT)/核因子-κB(NF-κB)通路相关蛋白磷酸化磷脂酰肌醇3激酶(P-PI3K)、磷酸化蛋白激酶B(P-AKT)、磷酸化核因子κB抑制因子α(P-IκBα)、核内核因子κB p65(Nuclear NF-κB p65)、胞质核因子κB p65(Cytoplasm NF-κB p65)的表达。[结果]体外实验结果表明SBT能显著促进EtOH损伤的GES-1细胞增殖,对细胞无明显毒性,明显提高细胞在EtOH中的活力。体内实验结果表明SBT预处理后可显著改善EtOH诱导的GU模型小鼠的体质量及进食量(P<0.05);减轻病理组织学损伤;降低小鼠血清炎性标志物IL-1β、TNF-α和IL-6的表达(P<0.01),升高IL-10的表达(P<0.01);降低小鼠胃黏膜组织ROS的水平(P<0.01),升高GSH-Px的水平(P<0.01),对MDA及NO影响则不显著;显著降低P-PI3K/PI3K、P-AKT/AKT和Nuclear NF-κB [Objective]This study aims to investigate the prophylactic and therapeutic effects of Sibai Decoction(SBT)on gastric ulcers(GU)induced by ethanol(EtOH)in mice,and to explore its underlying molecular mechanisms,thus providing a theoretical basis for its clinical application in treating GU.[Methods]In vitro,five groups of mice were administered with high,medium,and low doses of SBT for 7 days,after which blood was collected to separate serum for the establishment of a serological research model.Human gastric epithelial cells(GES-1)were cultured in media containing drug-serum and treated with 0.8 mol/L EtOH for 4 hours after entering the G0 phase to establish a GES-1 cell injury model.Cell viability and the effects of SBT-containing serum on cell proliferation were measured using the Cell Counting Kit-8(CCK8)method.In vivo,60 mice were divided into six groups evenly and pre-treated with SBT before EtOH induction to develop a gastric mucosal injury model.The effects of SBT on the mouse weight and food intake were observed.Histopathological changes in the gastric mucosa were assessed by Hematoxylin and Eosin(HE)staining.Levels of inflammatory markers,including Interleukin 6(IL-6),Interleukin 1 Beta(IL-1β),Tumor Necrosis Factor Alpha(TNF-α),and Interleukin 10(IL-10)protein expression levels,were measured in the serum using Enzyme-Linked Immunosorbent Assay(ELISA).Gastric mucosal levels of Malondialdehyde(MDA),Reactive Oxygen Species(ROS),Glutathione Peroxidase(GSH-Px),and Nitric Oxide(NO)were assessed using biochemical kits.The expression of proteins involved in the PI3K/AKT/NF-κB pathway,including Phosphorylated Phosphatidylinositol 3-Kinase(P-PI3K),Phosphorylated Protein Kinase B(P-AKT),Phosphorylated Nuclear factor kappa-B inhibitor alpha(P-IκBα),Nuclear Nuclear Factor Kappa B p65(Nuclear NF-κB p65),and Cytoplasmic Nuclear Factor Kappa B p65(Cytoplasm NF-κB p65),were analyzed by Western blot.[Results]In vitro experiments demonstrated that SBT significantly promoted the proliferation of EtOH-damaged GES-1 cell

关 键 词：四白汤 乙醇 胃溃疡 抗氧化应激 抗炎 

分 类 号：R285.5[医药卫生—中药学]