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

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 anti-inflammatory 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 43.76 μmol/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.