基于网络药理学和体外实验验证大蒜含硫化合物蒜氨酸抑制慢性气道炎症的作用机制研究  

Mechanism of action about garlic sulfur-containing compound alliin in inhibiting chronic airway inflammation based on network pharmacology and in vitro experiments

作　　者：王秋雨 李新霞[1] 兰怡[3] 毛新民 马红梅[2,5] WANG Qiu-yu;LI Xin-xia;LAN Yi;MAO Xin-min;MA Hong-mei(College of Pharmacy,Xinjiang Medical University,Urumqi 830011;College of Traditional Chinese Medicine,Xinjiang Medical University,Urumqi 830011;Affiliated Traditional Chinese Medicine Hospital of Xinjiang Medical University,Urumqi 830011;Key Laboratory of High Incidence Diseases in Xinjiang,Ministry of Education,Urumqi 830011;Key Discipline of Integrated Traditional and Western Medicine in Xinjiang Uygur Autonomous Region,Urumqi 830011)

机构地区：[1]新疆医科大学药学院,乌鲁木齐830011 [2]新疆医科大学中医学院,乌鲁木齐830011 [3]新疆医科大学附属中医医院,乌鲁木齐830011 [4]新疆地区高发疾病研究教育部重点实验室,乌鲁木齐830011 [5]新疆维吾尔自治区中西医结合重点学科,乌鲁木齐830011

出　　处：《中南药学》2023年第5期1170-1176,共7页Central South Pharmacy

基　　金：新疆维吾尔自治区自然科学基金面上项目(No.2020D01C162);2020年新疆维吾尔自治区研究生创新项目(No.XJ2020G176);新疆维吾尔自治区重大科技专项项目(No.2022A03007-2)。

摘　　要：目的通过网络药理学、分子对接模拟和体外细胞实验探讨蒜氨酸治疗慢性气道炎症的相关作用机制。方法运用数据库整理分析蒜氨酸与疾病共同靶点,进行KEGG通路富集分析,构建可视化药物-靶点-通路-疾病网络图。利用分子对接技术模拟获得相关通路蛋白与蒜氨酸结合能。通过MTT实验检测蒜氨酸对16HBE细胞活性的影响,并利用流式细胞术观察蒜氨酸对细胞氧化应激的保护作用,采用ELISA法检测蒜氨酸对细胞上清液中IL-6、TNF-α表达的影响、Western blot法检测各组中IL-6、TGF-β1、PPARα蛋白表达水平。结果蒜氨酸与疾病关键潜在靶点有EGFR、MMP9、AKT1、MAPK1、PPARα等,KEGG通路富集分析得到主要信号通路有MAPK信号通路和PPAR信号通路等,分子对接实验中蒜氨酸与MMP9、AKT1、STAT、PPARα结合能稳定,小于-20.92 kJ·mol^(-1)。体外细胞实验结果表明蒜氨酸对细胞无明显毒性作用,结果显示蒜氨酸可以明显降低炎症状态下细胞内过多的活性氧水平,上调PPARα表达,并显著下调炎症因子IL-6、TNF-α与TGF-β1的表达。结论蒜氨酸能改善因香烟烟雾与脂多糖刺激下16HBE细胞的氧化应激和炎症反应,可能通过调控PPARα信号通路改善细胞炎症损伤状态,从而恢复细胞内正常的抗氧化、炎症反应。Objective To determine the mechanism of action of alliin for chronic airway inflammation by network pharmacology,molecular docking and in vitro cell experiments.Methods The common targets of alliin and disease were analyzed by database collection,and the enrichment analysis of KEGG pathway was conducted to construct the visual drug-target-pathway-disease network diagram.The binding energy of related pathway proteins and alliin was obtained by molecular docking technique.MTT was used to detect the effect of alliin on the activity of 16HBE cells,flow cytometry was used to detect the effect of alliin on cell oxidative stress,ELISA was used to detect the effect of alliin on IL-6 and TNF-αexpression in the cell supernatant,and Western blot was used to detect the expression levels of IL-6,TGF-β1 and PPARαproteins in each group.Results Alliin and the key potential targets of disease included EGFR,MMP9,AKT1,MAPK1,and PPARα,etc.The enrichment analysis of KEGG pathway shows that the main signaling pathways included MAPK signaling pathway and PPAR signaling pathway,etc.The binding energy of alliin and MMP9,AKT1,STAT,and PPARαwas stable in the molecular docking,less than-20.92 kJ·mol^(-1).The in vitro cellular assays showed that alliin had no significant toxic effect on cells.Alliin significantly reduced the level of excessive intracellular reactive oxygen species(ROS)in inflammatory states,upregulated PPARαexpression,and significantly downregulated the expression of inflammatory factors IL-6,TNF-αand TGF-β1.Conclusion Alliin may ameliorate the oxidative stress and inflammatory responses in 16HBE cells stimulated by cigarette smoke and lipopolysaccharide,and may improve the state of cellular inflammatory damage by modulating the PPARαsignaling pathway,thereby restoring the normal intracellular antioxidant,and inflammatory responses.

关 键 词：16HBE细胞 网络药理学 分子对接 活性氧 炎症因子 蒜氨酸 

分 类 号：R285[医药卫生—中药学]