基于网络药理学和体外实验研究亚麻木酚素抗轮状病毒的作用机制  

Mechanism of flax lignans against rotavirus based on network pharmacology and in vitro experiments

作　　者：周玉晶 杨芷胭 余润宇 周芸湄 江小英 余俊贤 赵文昌 宋丽军 ZHOU Yujing;YANG Zhiyan;YU Runyu;ZHOU Yunmei;JIANG Xiaoying;YU Junxian;ZHAOWenchang;SONG Lijun(School of Pharmacy,Guangdong Medical University,Dongguan 523808,China;Guangdong Key Laboratory for Research and Development of Natural Drugs,Guangdong Medical University,Zhanjiang 524023,China)

机构地区：[1]广东医科大学药学院,广东东莞523808 [2]广东医科大学天然药物研发重点实验室,广东湛江524023

出　　处：《现代药物与临床》2024年第11期2760-2770,共11页Drugs & Clinic

基　　金：国家自然科学基金面上项目(81473401,81973548);广东省高校创新团队项目(2022KCXTD011);湛江市科技计划项目(2021B01139);广东医科大学学科重点项目(4SG23006G)。

摘　　要：目的基于网络药理学、分子对接和体外细胞实验探讨亚麻木酚素抗轮状病毒的作用机制。方法利用TCMSP、SwissTarget Prediction、PharmMapper数据库收集亚麻木酚素的作用靶点,利用GeneCards、OMIM、DisGeNET数据库获取轮状病毒的作用靶点。构建韦恩图并获得交集靶点,并利用Metascape数据库进行基因本体(GO)和京都基因与基因组百科全书(KEGG)富集分析。利用Pymol、AutoDock软件进行分子对接验证。采用轮状病毒感染MA104细胞模型研究亚麻木酚素的抗轮状病毒吸附、直接抑制轮状病毒和抗轮状病毒生物合成作用。测定细胞丙二醛(MDA)、谷胱甘肽(GSH)含量表达,研究亚麻木酚素对轮状病毒引起的氧化应激的影响。利用RT-qPCR和Western blotting探讨亚麻木酚素对轮状病毒感染的磷酸肌醇3-激酶(PI3K)/蛋白激酶B(Akt)/叉头框蛋白O1(FoxO1)信号通路的影响。结果通过网络药理学分析,共获取369个亚麻木酚素靶点,445个轮状病毒靶点,38个交集靶点,KEGG富集分析获得144条富集通路。分子对接表明靶蛋白受体与亚麻木酚素配体可以稳定结合。实验表明亚麻木酚素具有抗轮状病毒生物合成作用,无抗轮状病毒吸附和直接抑制轮状病毒作用。与模型组相比,亚麻木酚素药物2.0、4.0μmol/L组MDA显著含量下降、GSH含量升高(P<0.05、0.01)。RT-qPCR和Western blotting结果显示,亚麻木酚素可以上调PI3K/Akt/FoxO1信号通路中相关基因和蛋白的表达,降低VP6蛋白表达(P<0.05、0.01、0.001)。结论亚麻木酚素可以通过PI3K/Akt/FoxO1信号通路抑制轮状病毒诱导的氧化应激,从而发挥抗轮状病毒作用。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 4.0μmol/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.

关 键 词：亚麻木酚素 轮状病毒 氧化应激 网络药理学 分子对接 磷酸肌醇3-激酶/蛋白激酶B/叉头框蛋白O1信号通路 

分 类 号：R285[医药卫生—中药学]