基于GES-1细胞模型和鼠胃类器官模型的金红片功效物质基础研究

Study on Functional Substance Basis of Jinhong Tablet Based on GES-1 Cell Model and Mouse Gastric Organoid Model

  • 摘要:
    目的 探究金红片治疗慢性浅表性胃炎(Chronic superficial gastritis, CSG)的功效物质基础。
    方法 采用脂多糖(Lipopolysaccharide, LPS)诱导人胃黏膜上皮细胞(Human gastric epithelial cell, GES-1)炎症模型、LPS诱导鼠胃类器官炎症模型和乙醇诱导GES-1细胞氧化损伤模型,分别探讨金红片的抗炎、抗氧化作用及功效物质基础。采用MTS法检测细胞增殖活性;qPCR检测细胞及胃类器官中肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)和白细胞介素-8(IL-8)的mRNA相对表达;试剂盒检测细胞中超氧化物歧化酶(SOD)、丙二醛(MDA)和活性氧(ROS)的含量。
    结果 金红片及其10个潜在功效成分显著降低LPS诱导的GES-1细胞炎症模型和胃类器官炎症模型中炎症相关基因TNF-α、IL-1β、IL-6和IL-8的含量表达,推测这10个成分是金红片发挥抗炎作用的功效物质基础。金红片及其11个潜在功效成分明显降低MDA和ROS含量,提高SOD的活性,推测这11个成分是金红片发挥抗氧化作用的功效物质基础。
    结论 通过体外细胞及胃类器官实验初步确定别隐品碱、延胡索碱、脱氢紫堇碱、盐酸巴马汀、绿原酸、木香烃内酯、芦丁、槲皮苷、去氢木香内酯、四氢黄连碱、异绿原酸B、川楝素、原阿片碱和槲皮素是金红片治疗CSG的功效物质基础,为金红片质量标准提升积累科学依据。

     

    Abstract:
    OBJECTIVE To explore the functional substance basis of Jinhong Tablet in the treatment of chronic superficial gastritis (CSG).
    METHODS Three different models were constructed to investigate the anti-inflammatory and antioxidant effects, functional material basis of Jinhong Tablet: inflammatory model in human gastric epithelial cells (GES-1) induced by lipopolysaccharide (LPS), LPS-induced inflammatory model in mouse gastric organoids, and ethanol-induced oxidative damage model in GES-1 cells. MTS assay was performed to detect cell proliferation activity; qPCR was applied to measure the relative mRNA expression of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), interleukin-6 (IL-6), and interleukin-8 (IL-8) in cells and gastric organoids; and the levels of superoxide dismutase (SOD), malondialdehyde (MDA), and reactive oxygen species (ROS) in cells were detected.
    RESULTS Jinhong Tablet and 10 functional components significantly reduced the relative expression of inflammation-related genes TNF-α, IL-1β, IL-6, and IL-8 in LPS-induced GES-1 cells and gastric organoids, suggesting that these 10 components are the functional substance basis for the anti-inflammatory effects of Jin Hong Tablet. Jin Hong Tablet and 11 functional components markedly decreased the levels of MDA and ROS and increased the activity of SOD, indicating that these 11 components were the functional substance basis of the antioxidant effects of Jinhong Tablet.
    CONCLUSION Through in vitro cell and gastric organoid experiments, it has been preliminarily determined that allocryptopine, corydaline, dehydrocorydaline, palmatine hydrochloride, chlorogenic acid, costunolide, rutin, quercitrin, dehydrocostus lactone, tetrahydrocoptisine, isochlorogenic acid B, toosendanin, protopine, and quercetin are the functional material basis of Jinhong Tablet in treating CSG, accumulating scientific evidence for the enhancement of the quality standards of Jinhong Tablet.

     

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