基于Tim-3/aletin-9信号通路探究三仁汤对Hp相关性脾胃湿热证胃炎症状改善及巨噬细胞极化的影响  

Exploring the Effects of San Ren Decoction on the Improvement of Symptoms and Macrophage Polarization of Hp-Associated Gastritis with Spleen and Stomach Damp Heat Syndrome Based on Tim-3/aletin-9 Signaling Pathway

作　　者：江望[1] 田生望[1] 贺单[1] 陶红[1] JIANG Wang;TIAN Shengwang;HE Dan;TAO Hong(The Second Affiliated Hospital of Chongqing Medical University,Chongqing 400010,China)

机构地区：[1]重庆医科大学附属第二医院,重庆400010

出　　处：《世界科学技术-中医药现代化》2025年第2期484-491,共8页Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology

基　　金：重庆市科技局面上项目(cstc2019jcyj-msxmX0178):不同脾胃湿热证候模型评价与诊断要素的研究,负责人:江望。

摘　　要：目的本研究旨在探索三仁汤通过Tim-3/Galectin-9信号途径对幽门螺旋杆菌(Helicobacter pylori,Hp)相关性胃炎(HAG)脾胃湿热证巨噬细胞极化的作用。方法72只Wistar大鼠随机分为空白组、模型组、四联疗法组(奥美拉唑2 mg/kg+阿莫西林100 mg/kg+克拉霉素50 mg/kg+胶体果胶铋胶囊35 mg/kg)、三仁汤低剂量组(3 g/kg)、三仁汤中剂量组(7.5 g/kg)、三仁汤高剂量组(15 g/kg),每组12只。采用复合因素(劳倦+饮食+苦寒药+环境+生物因子(Hp菌液))构建HAG脾胃湿热证大鼠模型,并按组别持续给药21 d。实验中观察大鼠的基本状况;快速尿素酶实验检测Hp定植;利用HE染色观察胃黏膜炎症情况;透射电镜观察胃黏膜超微结构;ELISA法测定血清IFN-γ、TNF-α、IL-4、IL-10水平;免疫组化检测胃黏膜CD68的表达;流式细胞术检测巨噬细胞F4/80^(+)CD86^(+)和F4/80^(+)CD206^(+)亚型;RT-qPCR检测胃黏膜组织MCP-1、CD80、CD86、IL-4、CD206、CD163 mRNA表达;Western blot检测Tim-3、Galectin-9蛋白在胃黏膜组织中的表达。结果模型组胃黏膜组织病理较空白组明显损伤,且Hp定植率、血清IFN-γ和TNF-α、胃黏膜组织CD68、Tim-3、Galectin-9蛋白表达及MCP-1、CD80、CD86、CD163 mRNA表达较空白组显著升高(P<0.01);三仁汤高剂量干预显著改善了模型大鼠胃黏膜组织损伤,显著降低了Hp定植率,并抑制了血清IFN-γ和TNF-α、胃黏膜组织CD68、Tim-3、Galectin-9蛋白表达、胃黏膜组织MCP-1、CD80、CD86、CD163mRNA表达(P<0.05)。结论三仁汤对大鼠HAG脾胃湿热证症状有改善作用,同时可抑制炎症反应并调节胃黏膜巨噬细胞的极化,其作用机制可能与调节Tim-3/Galectin-9途径有关。Objective To explore the effect of San Ren decoction on macrophage polarization in Helicobacter pylori(Hp)-associated gastritis(HAG)with spleen and stomach damp heat syndrome through Tim-3/Galectin-9 signaling pathway.Methods Seventy-two Wistar rats were randomly divided into control group,model group,quadruple therapy group(omeprazole 2 mg/kg+amoxicillin 100 mg/kg+clarithromycin 50 mg/kg+colloidal pectin secret capsule 35 mg/kg),San Ren decoction low dose group(3 g/kg),San Ren decoction medium dose group(7.5 g/kg),San Ren decoction high dose group(15 g/kg),12 animals in each group.The rat model of HAG spleen and stomach damp heat syndrome was constructed by composite factors(fatigue+diet+bitter cold medicine+environment+biological factors(Hp bacterial solution)),and the drugs were continuously administered for 21 days according to the groups.The basic conditions of rats were observed in the experiment.Hp colonization was detected by rapid urease test.And gastric mucosal inflammation was observed using HE staining.A transmission electron microscope was used to observe the ultrastructure of the gastric mucosa.The levels of IFN-γ,TNF-α,IL-4,IL-10 in serum were measured by ELISA.Gastric mucosa expressed CD68 by immunohistochemistry.Subtypes of macrophages F4/80^(+)CD86^(+)and F4/80^(+)CD206^(+)were detected by flow cytometry.RT-qPCR was performed to detect the mRNA expression of MCP-1,CD80,CD86,IL-4,CD206,and CD163 in gastric mucosa.Western blot was used to detect the expression of Tim-3 and Galectin-9 proteins in gastric mucosal tissues.Results The histopathology of gastric mucosa in the model group was significantly damaged compared with that of the control group,and the rate of Hp colonization,serum IFN-γ and TNF-α,protein expression of CD68,Tim-3 and Galectin-9 in gastric mucosa tissues,and mRNA expression of MCP-1,CD80,CD86,and CD163 were significantly higher compared with that of the control group(P<0.01).The high-dose intervention of San Ren decoction significantly improved the gastric mucosal tissue inju

关 键 词：幽门螺旋杆菌相关性胃炎 脾胃湿热证 三仁汤 Tim-3/Galectin-9通路 巨噬细胞 

分 类 号：R285.5[医药卫生—中药学]