基于高分辨质谱和网络药理学探究南昆山毛叶红茶的抗炎机理

Exploring Analytics of Mechanisms Behind the Anti-inflammatory Activity of Black Tea from Camellia ptilophylla Chang Based on Highresolution Mass Spectrum and Network Pharmacology

目的:探究南昆山毛叶红茶抗炎功效发挥的化学基础及分子机制。方法:采用超高效液相色谱-四极杆-静电场轨道阱高分辨质谱(ultra-high performance liquid chromatography-quadrupole orbitrap high resolution-mass spectrometry,UPLC-Q-Orbitrap HRMS)技术分析其醇提物活性组分,通过网络药理学预测其抗炎活性组分、作用靶点和信号通路,利用分子对接技术验证抗炎活性组分与靶点的相互作用能力,采用体外炎症细胞模型验证活性组分对炎性介质一氧化氮的抑制作用。结果:从南昆山毛叶红茶中鉴定出213种活性组分,并筛选出花青素、柚皮素、木犀草素、槲皮素等核心抗炎多酚类组分;收集到潜在抗炎靶点493个,并预测了肿瘤坏死因子(tumor necrosis factor,TNF)、白细胞介素-6(interleukin-6,IL-6)等9个关键靶点和磷脂酰肌醇3-激酶/蛋白激酶B(phosphatidylinositol 3-kinase/protein kinase B,PI3K-Akt)、丝裂原活化蛋白激酶(mitogen-activated protein kinase,MAPK)等多条信号通路;初步证明11种抗炎活性组分、特别是木犀草素等9种多酚类物质能与主要靶点稳定结合,且木犀草素、槲皮素能够明显抑制脂多糖诱导的小鼠巨噬细胞RAW264.7产生一氧化氮(nitric oxide,NO),半抑制浓度分别为18.91、43.76μmol/L。结论:南昆山毛叶红茶通过木犀草素、槲皮素等多酚类物质作用于肿瘤坏死因子等多个靶点和丝裂原活化蛋白激酶等多条通路发挥抗炎活性。本研究为深入揭示南昆山毛叶茶的抗炎活性提供了理论研究基础。

Objective:In order to explore the mechanisms behind the anti-inflammatory potentials of black tea from Camellia ptilophylla Chang(C.ptilophylla)(BTCP).Method:The bioactive components of the methanol extracts of BTCP were firstly detected via ultra-high performance liquid chromatography-quadrupole orbitrap high-resolution mass spectrometry(UPLC-Q-Orbitrap HRMS).Additionally,the bioactive components,targets,and signal pathways involved in the antiinflammatory effects of BTCP were predicted by network pharmacology(NP).Then,the interactions between the bioactive components and the targets were evaluated using molecular docking(MD).Finally,Inflammatory cell model was used to verify the inhibitory effects of the bioactive components on nitric oxide in vitro,a pro-inflammatory cytokine.Result:A total of 213 active components were identified from BTCP,and the core anti-inflammatory polyphenols such as anthocyanin,naringin,luteolin and quercetin were screened.In addition,493 potential anti-inflammatory targets were collected,and tumor necrosis factor(TNF),interleukin(IL-6)were predicted.IL-6 and other nine key targets and phosphatidylinositol 3-kinase/protein kinase B,PI3K-Akt,mitogen-activated protein kinase(MAPK)and other signaling pathways.Preliminary results showed that eleven anti-inflammatory active components,especially nine polyphenols such as luteolin,could stably bind to the main target,and luteolin and quercetin could significantly inhibit the production of nitric oxide(NO)in mouse macrophage RAW264.7 induced by lipaccharide.The semi-inhibitory concentrations were 18.91 and 43.76μmol/L,respectively.Conclusion:Polyphenol compounds such as luteolin in BTCP would exert remarkable anti-inflammatory activity through the regulation on multiple targets including TNF and IL-6 and various pathways such as PI3K-Akt and MAPK.The present study provides a theoretical basis for further elucidation of the anti-inflammatory potentials of C.ptilophylla.