目的 基于網(wǎng)絡(luò)藥理學(xué)、分子對接和體外細(xì)胞實(shí)驗(yàn)探討亞麻木酚素抗輪狀病毒的作用機(jī)制。方法 利用TCMSP、SwissTarget Prediction、PharmMapper數(shù)據(jù)庫收集亞麻木酚素的作用靶點(diǎn)，利用GeneCards、OMIM、DisGeNET數(shù)據(jù)庫獲取輪狀病毒的作用靶點(diǎn)。構(gòu)建韋恩圖并獲得交集靶點(diǎn)，并利用Metascape數(shù)據(jù)庫進(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Pymol、AutoDock軟件進(jìn)行分子對接驗(yàn)證。采用輪狀病毒感染MA104細(xì)胞模型研究亞麻木酚素的抗輪狀病毒吸附、直接抑制輪狀病毒和抗輪狀病毒生物合成作用。測定細(xì)胞丙二醛（MDA）、谷胱甘肽（GSH）含量表達(dá)，研究亞麻木酚素對輪狀病毒引起的氧化應(yīng)激的影響。利用RT-qPCR和Western blotting探討亞麻木酚素對輪狀病毒感染的磷酸肌醇3-激酶（PI3K）/蛋白激酶B（Akt）/叉頭框蛋白O1（FoxO1）信號通路的影響。結(jié)果 通過網(wǎng)絡(luò)藥理學(xué)分析，共獲取369個(gè)亞麻木酚素靶點(diǎn)，445個(gè)輪狀病毒靶點(diǎn)，38個(gè)交集靶點(diǎn)，KEGG富集分析獲得144條富集通路。分子對接表明靶蛋白受體與亞麻木酚素配體可以穩(wěn)定結(jié)合。實(shí)驗(yàn)表明亞麻木酚素具有抗輪狀病毒生物合成作用，無抗輪狀病毒吸附和直接抑制輪狀病毒作用。與模型組相比，亞麻木酚素藥物2.0、4.0 μmol/L組MDA顯著含量下降、GSH含量升高（P＜0.05、0.01）。RT-qPCR和Western blotting結(jié)果顯示，亞麻木酚素可以上調(diào)PI3K/Akt/FoxO1信號通路中相關(guān)基因和蛋白的表達(dá)，降低VP6蛋白表達(dá)（P＜0.05、0.01、0.001）。結(jié)論 亞麻木酚素可以通過PI3K/Akt/FoxO1信號通路抑制輪狀病毒誘導(dǎo)的氧化應(yīng)激，從而發(fā)揮抗輪狀病毒作用。

Objective To investigate the effect and mechanism of flax lignans against rotavirus based on network pharmacology, molecular docking and in vitro cellular assays. Methods TCMSP, SwissTargetPrediction, PharmMapper databases were used to collect the targets of flax lignans. GeneCards, OMIM, DisGeNET databases were used to obtain the targets of rotavirus. Venn diagram was constructed to obtain intersection targets, and GO and KEGG enrichment analysis was performed using Metascape database. Molecular docking validation was performed using Pymol, AutoDock software. The inhibition of rotavirus attachment, direct inhibition of rotavirus and inhibition of rotavirus replication effects of flax lignans were studied using the rotavirus -infected MA104 cell model. The effects of flax lignans on rotavirus-induced oxidative stress were investigated by measuring the content expression of MDA and GSH oxidation indicators. RT-qPCR and Western blotting were used to explore the effect of flax lignans on PI3K/Akt/FoxO1 signaling pathway in rotavirus infection. Results A total of 369 flax lignans targets, 445 rotavirus targets, and 38 crossover targets were obtained by network pharmacological analysis, and 144 enriched pathways were obtained by KEGG enrichment analysis. Molecular docking showed that the target protein receptors could bind stably to the flax lignans. The results showed that flax lignans had anti- rotavirus replication effect without anti- rotavirus adsorption and direct inhibition of rotavirus. Compared with the model group, the content of MDA were decreased, and GSH were increased in the flax lignans 2.0 and 4.0 μmol/L group (P < 0.05, 0.01). RT-qPCR and Western blotting results showed that flax lignans could upregulate the expression of related genes and proteins in the PI3K/Akt/FoxO1 signaling pathway, and decreased VP6 (P < 0.05, 0.01, 0.001). Conclusion Flax lignans can exert anti-rotavirus effects by inhibiting rotavirus -induced oxidative stress through the PI3K/Akt/FoxO1 signaling pathway.

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國家自然科學(xué)基金面上項(xiàng)目(81473401,81973548);廣東省高校創(chuàng)新團(tuán)隊(duì)項(xiàng)目(2022KCXTD011);湛江市科技計(jì)劃項(xiàng)目(2021B01139);廣東醫(yī)科大學(xué)學(xué)科重點(diǎn)項(xiàng)目(4SG23006G)