目的 基于網(wǎng)絡(luò)藥理學(xué)和分子對接技術(shù)探討麻黃細(xì)辛附子湯治療變應(yīng)性鼻炎（AR）的作用機(jī)制，并結(jié)合體外細(xì)胞實(shí)驗(yàn)進(jìn)行驗(yàn)證。方法 運(yùn)用TCMSP平臺獲取麻黃細(xì)辛附子湯的活性成分及作用靶點(diǎn)，通過GeneCards、OMIM、PharmGKB、TTD和DrugBank數(shù)據(jù)庫檢索AR疾病靶點(diǎn)。利用STRING平臺構(gòu)建蛋白質(zhì)相互作用（PPI）網(wǎng)絡(luò)。通過R語言進(jìn)行基因本體（GO）和京都基因與基因組百科全書（KEGG）富集分析。利用AutoDock對關(guān)鍵化合物與核心靶基因進(jìn)行對接驗(yàn)證。體外培養(yǎng)人鼻黏膜上皮細(xì)胞（HNEpC），使用酶聯(lián)免疫吸附（ELISA）、流式細(xì)胞術(shù)、實(shí)時熒光定量PCR（qRTPCR）的方法驗(yàn)證麻黃細(xì)辛附子湯對炎癥因子、細(xì)胞凋亡、部分關(guān)鍵靶點(diǎn)表達(dá)的影響。結(jié)果 經(jīng)篩選得到麻黃細(xì)辛附子湯活性成分62個，治療AR靶點(diǎn)99個。PPI核心靶點(diǎn)包括前列腺素G/H合成酶2（PTGS2）、絲裂原活化蛋白激酶3（MAPK3）、基質(zhì)金屬蛋白酶-9（MMP-9）等。KEGG富集分析涉及與免疫炎癥相關(guān)信號通路，與細(xì)胞增殖、凋亡相關(guān)信號通路，GO富集分析涉及對脂多糖反應(yīng)等生物過程，細(xì)胞因子受體結(jié)合、細(xì)胞因子活性等分子功能。分子對接結(jié)果表明，活性成分與靶基因緊密結(jié)合。細(xì)胞實(shí)驗(yàn)結(jié)果顯示，與模型組比較，麻黃細(xì)辛附子湯可以減少炎癥因子白細(xì)胞介素-6（IL-6）、白細(xì)胞介素-8（IL-8）、腫瘤壞死因子-α（TNF-α）分泌（P＜0.05、0.01），抑制細(xì)胞凋亡（P＜0.01），降低PTGS2、MAPK3 mRNA表達(dá)、升高M(jìn)MP-9 mRNA表達(dá)（P＜0.05、0.01）。結(jié)論 麻黃細(xì)辛附子湯可以通過多種活性成分作用于多靶點(diǎn)、多通路，調(diào)控炎癥反應(yīng)、免疫功能、細(xì)胞凋亡等途徑發(fā)揮治療AR的作用。

Objective To investigate the mechanism of action of Mahuang Xixin Fuzi Decoction in treatment of allergic rhinitis (AR) based on network pharmacology and molecular docking techniques, and to validate it in combination with in vitro cellular experiments. Methods The TCMSP platform was used to obtain the active ingredients and action targets of Mahuang Xixin Fuzi Decoction, and the AR disease targets were searched by GeneCards, OMIM, PharmGKB, TTD and DrugBank databases. The protein-protein interaction (PPI) network was established by STRING platform. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis were performed by R language. AutoDock was used to verify the docking of key compounds with core target genes. Human nasal epithelial cells (HNEpC) were cultured in vitro, and the effects of Mahuang Xixin Fuzi Decoction on inflammatory factors, apoptosis, and expression of some key targets were verified using ELISA, flow cytometry, and qRT-PCR. Results A total of 62 active components of Mahuang Xixin Fuzi Decoction and 99 therapeutic AR targets were obtained. The core targets of PPI included PTGS2, MAPK3, MMP-9, etc. KEGG enrichment analysis involved signaling pathways related to immune inflammation, cell proliferation and apoptosis, GO enrichment analysis involved biological processes such as response to lipopolysaccharide, cytokine receptor binding, cytokine activity. The molecular docking results showed that the active ingredients were tightly bound to the target genes. The results of cellular experiment showed that compared with the model group, Mahuang Xixin Fuzi Decoction could reduce the secretion of inflammatory factors IL-6, IL-8 and TNF-α (P< 0.05, 0.01), inhibit apoptosis (P< 0.01), reduce PTGS2 and MAPK3 mRNA expression and elevate MMP-9 mRNA expression (P< 0.05, 0.01). Conclusion Mahuang Xixin Fuzi Decoction can act on multiple targets and pathways through multiple active ingredients to regulate inflammatory response, immune function, and apoptosis to exert therapeutic effects on AR.

R285.5

國家自然科學(xué)基金資助項(xiàng)目（81774375）