目的 采用網(wǎng)絡(luò)藥理學(xué)的方法預(yù)測氣滯胃痛顆粒治療胃炎主要活性成分的作用靶點及其作用機制，并采用細(xì)胞實驗對部分主要靶點進(jìn)行實驗驗證。方法 在中藥系統(tǒng)藥理學(xué)數(shù)據(jù)庫與分析平臺（TCMSP）和網(wǎng)絡(luò)藥理學(xué)在線數(shù)據(jù)庫（TCMID）中獲得氣滯胃痛顆粒組方中藥柴胡、白芍、枳殼、香附（炙）、延胡索（炙）、甘草（炙）的化學(xué)成分，并在TCMSP和PharmMapper數(shù)據(jù)庫中收集活性成分對應(yīng)的靶點，在藥物靶標(biāo)數(shù)據(jù)庫（TTD）和人類孟德爾遺傳數(shù)據(jù)庫（OMIM）數(shù)據(jù)庫中搜索與胃炎相關(guān)的蛋白和基因，建立胃炎靶點數(shù)據(jù)庫；通過DIP數(shù)據(jù)庫將成分靶點和疾病靶點進(jìn)行關(guān)聯(lián)，建立化合物-靶點-疾病網(wǎng)絡(luò)。將氣滯胃痛顆粒治療胃炎的相關(guān)靶點輸入STRING數(shù)據(jù)庫中構(gòu)建靶點之間的蛋白質(zhì)相互作用（PPI）關(guān)系；使用DAVID數(shù)據(jù)庫對靶點進(jìn)行京都基因與基因組百科全書（KEGG）通路富集分析，使用Cytoscape3.5.1軟件自帶的GlueGO插件進(jìn)行基因本體論（GO）生物功能注釋；通過人類基因和基因表型綜合數(shù)據(jù)庫（OMM）篩選炎癥相關(guān)細(xì)胞相關(guān)的基因和蛋白靶點，對氣滯胃痛顆粒治療胃炎的靶細(xì)胞進(jìn)行富集。體外細(xì)胞實驗以巨噬細(xì)胞RAW264.7為對象，采用1μg·mL^-1脂多糖（LPS）和0.2μg·mL^-1γ干擾素（IFN-γ）制備細(xì)胞模型，以含有最大無毒劑量氣滯胃痛顆粒內(nèi)容物的培養(yǎng)基處理RAW264.7模型細(xì)胞，采用實時熒光定量PCR（qRT-PCR）法測定各組RAW264.7細(xì)胞中環(huán)氧合酶2（COX-2）、誘導(dǎo)型一氧化氮合酶（iNOS）、白細(xì)胞介素6（IL-6）、腫瘤壞死因子α（TNF-α）mRNA表達(dá)。結(jié)果 網(wǎng)絡(luò)藥理學(xué)預(yù)測得到氣滯胃痛顆粒治療胃炎的主要潛在的活性成分25個，重要靶點20個，其中關(guān)鍵靶點包括COX-2、iNOS、過氧化物酶體增殖物激活受體γ（PPARγ）、絲裂原活化蛋白激酶14（MAPK-14）、表皮生長因子受體（EGFR）等，作用機制可能與調(diào)節(jié)TNF信號通路、NOD樣受體信號通路、VEGF信號通路等與胃炎密切相關(guān)的信號通路有關(guān)，其藥效作用主要表現(xiàn)為對炎癥、血管內(nèi)穩(wěn)態(tài)、免疫、中樞神經(jīng)及激素調(diào)節(jié)等生物過程的影響。氣滯胃痛顆粒對RAW264.7模型細(xì)胞中COX-2、iNOS、IL-6、TNF-α的mRNA表達(dá)具有顯著抑制作用。結(jié)論 氣滯胃痛顆?？赡苤饕ㄟ^修復(fù)胃黏膜、減輕炎癥損傷、消除感染等實現(xiàn)其治療胃炎作用。

Objective To predict the target and mechanism of Qizhi Weitong Granule in treating gastritis by network pharmacological method, and to verify some of the main targets by cell experiment.Methods The chemical components of Bupleuri Radix, Paeoniae Radix Alba, Aurantii Fructus, Cyperi Rhizoma, Corydalis Rhizoma and Glycyrrhizae Radix et Rhizoma were obtained from TCMSP and TCMID, and the corresponding target points of active components were collected from TCMSP and PharmaMapper databases, Search the drug target database (TTD) and the human Mendelian genetic database (OMIM) database for proteins and genes related to gastritis, and establish the gastritis target database. The component target and disease target are related through DIP database to establish a compound target disease network. Input the relevant targets of Qizhi Weitong Granule for gastritis into STRING database to build the protein interaction (PPI) relationship between the targets. The DAVID database was used to enrich and analyze the Kyoto encyclopedia of genes and genomes (KEGG) pathway of the target, and the Glue GO plug-in included in the Cytoscape 3.5.1 software was used to annotate the biological function of the gene ontology (GO). The gene and protein targets related to inflammation related cells were screened through the comprehensive database of human genes and gene phenotypes (OMM) to enrich the target cells of Qizhi Weitong Granule in treating gastritis. In vitro cell experiment, macrophage RAW264.7 was used as the object, and the cell model was prepared by 1 μg·mL^-1 lipopolysaccharide (LPS) and 0.2 μg·mL^-1 γ interferon (IFN-γ). RAW264.7 model cells were treated with the medium containing the largest non-toxic dose of Qizhi Weitong Granules. The mRNA expression of cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS), interleukin-6 (IL-6) and tumor necrosis factor α (TNF-α) in RAW264.7 cells in each group were determined by real-time fluorescent quantitative PCR (qRTPCR).Results The network pharmacology predicted that there were 25 main potential active components and 20 important targets of Qizhi Weitong Granule in treating gastritis, and key targets include COX-2, iNOS, peroxisome proliferator activated receptor γ (PPAR-γ) mitogen activated protein kinase 14 (MAPK-14), epidermal growth factor receptor (EGFR), etc. The action mechanism of Qizhi Weitong Granule in treating gastritis may be related to the regulation of TNF signal pathway, NOD like receptor signal pathway, VEGF signal pathway and other signal pathways closely related to gastritis. Its pharmacodynamic effect is mainly manifested in its influence on biological processes such as inflammation, vascular homeostasis, immunity, central nervous system and hormone regulation. The mRNA expression of COX-2, iNOS, IL-6, TNF-α in RAW264.7 model cells were significantly inhibited by Qizhi Weitong Granule.Conclusion Qizhi Weitong Granule may achieve its therapeutic effect on gastritis mainly by repairing gastric mucosa, alleviating inflammatory damage and eliminating infection.

R285.5