摘要
目的考察厚朴三物汤对乙醇诱导的急性胃溃疡大鼠的改善作用,并从胃脑相关角度探究厚朴三物汤改善胃溃疡大鼠的作用机制。方法采用UPLC-Q-Exactive-MS分析厚朴三物汤中的化学成分。SD大鼠随机分为对照组、模型组、奥美拉唑(30 mg/kg)组及厚朴三物汤低、中、高剂量(4.725、9.450、18.900 g/kg)组,建立乙醇致胃溃疡模型,采用苏木素-伊红(HE)染色观察大鼠胃黏膜损伤情况;ELISA测定大鼠胃组织中肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-6(interleukin-6,IL-6)、IL-1β、前列腺素E_(2)(prostaglandin E_(2),PGE_(2))、表皮生长因子(epidermal growth factor,EGF)和脑组织中5-羟色胺(5-hydroxytryptamine,5-HT)含量;比色法测定大鼠胃组织超氧化物歧化酶(superoxide dismutase,SOD)和过氧化氢酶(catalase,CAT)活性;微板法测定大鼠胃组织丙二醛(malondialdehyde,MDA)和还原型谷胱甘肽(glutathione,GSH)含量;运用转录组学测序(transcriptome sequencing,RNA-Seq)和生物信息学技术筛选厚朴三物汤中剂量组治疗胃溃疡大鼠可能干预的信号通路。结果初步鉴定出厚朴三物汤共71个化学成分,包括黄酮类26个、鞣质类6个、蒽醌及蒽酮类14个、苯丙素类15个等。与模型组比较,厚朴三物汤能在不同程度上减轻炎症、改善氧化应激和增加胃黏膜因子水平,进而修复胃溃疡大鼠的胃黏膜损伤,其中厚朴三物汤高剂量组效果最优,能显著降低胃溃疡大鼠炎症因子TNF-α、IL-6和IL-1β水平(P<0.001),增加胃黏膜保护因子PGE_(2)和EGF含量(P<0.01),升高胃溃疡大鼠胃组织中CAT活性和GSH水平(P<0.01),降低MDA水平(P<0.01)。给予厚朴三物汤干预后,胃溃疡大鼠脑组织中5-HT含量下降趋势均得到不同程度的逆转(P<0.05)。RNA-Seq分析结果显示,模型组相较于厚朴三物汤中剂量组脑组织中上调基因113个,下调基因59个。基因本体(gene ontology,GO)功能分析发现厚朴三物汤治疗后差异基因主要富集在离子转运、细胞骨架与胞质膜、突触受体传递等方面。京都基因与基因组百科全书(Kyoto encyclopedia of genes and genomes,KEGG)通路分析发现厚朴三物汤治疗后主要富集在NOD样受体信号通路、神经活性配体-受体相互作用和人乳头瘤病毒感染等代谢通路,说明其可能是厚朴三物汤干预胃溃疡脑神经从而改善胃黏膜损伤的重要通路。结论厚朴三物汤能明显改善胃溃疡大鼠的胃黏膜损伤,其作用机制与胃、脑的炎症反应与代谢紊乱有关。从胃脑相关角度出发,探索厚朴三物汤通过调节脑神经系统改善胃溃疡大鼠胃黏膜损伤的作用,为经典名方厚朴三物汤的抗胃溃疡作用机制研究提供新思路。
Objective To investigate the effect of Houpo Sanwu Decoction(厚朴三物汤)on ethanol-induced acute gastric ulcer in rats,and explore the mechanism of Houpo Sanwu Decoction on improving gastric ulcer in rats from the perspective of gastric-brain.Methods UPLC-Q-Exactive-MS method was used to analyze the chemical components in Houpo Sanwu Decoction.SD rats were randomly divided into control group,model group,omeprazole(30 mg/kg)group,and Houpo Sanwu Decoction low-,medium-,and high-dose(4.725,9.450,18.900 g/kg)groups.After administration,an ethanol-induced gastric ulcer model was prepared,and the damage to the gastric mucosa of rats was observed using hematoxylin eosin(HE)staining;ELISA was used to determine the levels of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),IL-1β,prostaglandin E_(2)(PGE_(2)),epidermal growth factor(EGF)in gastric tissue and 5-hydroxytryptamine(5-HT)level in brain tissue of rats;Colorimetric method was used to measure the activities of superoxide dismutase(SOD)and catalase(CAT)in gastric tissue of rats;Microplate method was used to determine the contents of malondialdehyde(MDA)and glutathione(GSH)in gastric tissue of rats;Transcriptome sequencing(RNA Seq)and bioinformatics techniques were used to screen for potential intervention signaling pathways in the treatment of gastric ulcer rats with the medium-dose group of Houpo Sanwu Decoction.Results A total of 71 chemical components were identified in Houpo Sanwu Decoction,including 26 flavonoids,six ellagitannins,14 anthraquinones and anthrones and 15 phenylpropanoids.Compared with model group,Houpo Sanwu Decoction could alleviate inflammation,improve oxidative stress and increase gastric mucosal factor levels to varying degrees,thereby repairing gastric mucosal damage in rats with gastric ulcer.Among them,high-dose group of Houpo Sanwu Decoction had the best effect,significantly reducing the levels of inflammatory factors TNF-α,IL-6 and IL-1βin rats with gastric ulcer(P<0.001),increasing the contents of gastric mucosal protective factors PGE_(2) and EGF(P<0.01),increasing CAT activity and GSH level in gastric tissue of rats with gastric ulcer(P<0.01),and reducing MDA level(P<0.01).After intervention with Houpo Sanwu Decoction,the decreasing trend of 5-HT content in brain tissue of rats with gastric ulcer was reversed to varying degrees(P<0.05).RNA-Seq analysis results showed that there were 113 up-regulated genes and 59 down-regulated genes in brain tissue of model group compared to the medium-dose group of Houpo Sanwu Decoction.Gene ontology(GO)functional analysis revealed that after treatment with Houpo Sanwu Decoction,differentially expressed genes were mainly enriched in ion transport,cytoskeleton and cytoplasmic membrane,and synaptic receptor transport.Kyoto encyclopedia of genes and genomes(KEGG)pathway analysis revealed that after treatment with Houpo Sanwu Decoction,it was mainly enriched in metabolic pathways such as NOD like receptor signaling pathway,neuroactive ligand receptor interaction,and human papillomavirus infection,indicating that it may be an important pathway for Houpo Sanwu Decoction to intervene in gastric ulcer brain nerves and improve gastric mucosal damage.Conclusion Houpo Sanwu Decoction can significantly improve gastric mucosal damage in rats with gastric ulcers,and its mechanism is related to the inflammatory response and metabolic disorders in the stomach and brain.This study explores the effect of Houpo Sanwu Decoction on improving gastric mucosal damage in rats with gastric ulcer by regulating the brain nervous system from the perspective of stomach brain related factors,providing new ideas for the study of the anti-gastric ulcer mechanism of the classic formula Houpo Sanwu Decoction.
作者
刘芳
张芮苑
沈才佳
付欣怡
宋姚
雷以珵
庞惠文
刘洋
陈晨
李杰
LIU Fang;ZHANG Ruiyuan;SHEN Caijia;FU Xinyi;SONG Yao;LEI Yichen;PANG Huiwen;LIU Yang;CHEN Chen;LI Jie(State Key Laboratory of Traditional Chinese Medicine Resources with Southwest Characteristics,School of Pharmacy,Chengdu University of Traditional Chinese Medicine,Chengdu 611137,China;School of Biomedical Sciences,University of Queensland,Brisbane 4072,Australia;Institute for Bioengineering and Nanotechnology,University of Queensland,Brisbane 4072,Australia;Center for Medical Research and Translation,The Fifth People’s Hospital/Second Clinical College of Medicine,Chengdu University of Traditional Chinese Medicine,Chengdu 611135,China;Institute of Herbal Genomics,Chengdu University of Traditional Chinese Medicine,Chengdu 611137,China)
出处
《中草药》
CAS
CSCD
北大核心
2024年第16期5525-5540,共16页
Chinese Traditional and Herbal Drugs
基金
四川省自然科学基金资助项目(2023NSFSC0661)
国家留学基金委西部地区人才培养特别项目(202208515025)。
关键词
厚朴三物汤
急性胃溃疡
胃脑相关
转录组学
胃黏膜保护
Houpo Sanwu Decoction
acute gastric ulcer
stomach-brain correlation
transcriptomics
gastric mucosal protection
