Metabolic profiling of Lantana camara L. using UPLC-MS/MS and revealing its inflammation-related targets using network pharmacology-based and molecular docking analyses

Abstract

Lantana camara L. is widely used in folk medicine for alleviation of inflammatory disorders, but studies that proved this folk use and that revealed the molecular mechanism of action in inflammation mitigation are not enough. Therefore, this study aimed to identify L. camara phytoconstituents using UPLC-MS/MS and explain their multi-level mechanism of action in inflammation alleviation using network pharmacology analysis together with molecular docking and in vitro testing. Fifty-seven phytoconstituents were identified in L. camara extract, from which the top hit compounds related to inflammation were ferulic acid, catechin gallate, myricetin and iso-ferulic acid. Whereas the most enriched inflammation related genes were PRKCA, RELA, IL2, MAPK 14 and FOS. Furthermore, the most enriched inflammation-related pathways were PI3K-Akt and MAPK signaling pathways. Molecular docking revealed that catechin gallate possessed the lowest binding energy against PRKCA, RELA and IL2, while myricetin had the most stabilized interaction against MAPK14 and FOS. In vitro cytotoxicity and anti-inflammatory testing indicated that L. camara extract is safer than piroxicam and has a strong anti-inflammatory activity comparable to it. This study is a first step in proving the folk uses of L. camara in palliating inflammatory ailments and institutes the groundwork for future clinical studies.

Figure 1

Study workflow diagram.

Figure 2

Doughnut charts showing the distributions % of the compound–target gene (C–T) interactions on L. camara constituents (a) and the identified inflammation-related proteins (b).

Figure 3

Compound–target–pathway network (compounds are represented in violet color, targets are presented in blue color and pathways are presented in red color).

Figure 4

(a) Gene Ontology analysis of inflammation targets determined by DAVID database. Biological processes, molecular functions and cellular components are represented by green, orange and blue bars, respectively. (b) Major BBID (green), BIOCARTA (orange) and KEGG (blue) pathways clusters generated from DAVID database. The significance of enrichment is indicated by log P-value with bar charts. Red lines represent the number of genes enriched by each term.

Figure 5

Bar chart showing (a) cytotoxicity (CC50 µg/mL), (b) effective anti-inflammatory concentrations (EAICS) (µg/mL) of L. camara extract and piroxicam, (c) TNF-α, IL-1β, INF-γ, IL-6 (fold change in gene expression) by L. camara extract and piroxicam (standard anti-inflammatory drug).

Figure 6

2D and 3D interaction diagrams of (a) catechin gallate in the active site of protein kinase C alpha type (PDB ID 4RA4) (b) catechin gallate in the active site of transcription factor p65 (PDB ID 3QXY) (c) catechin gallate in the active site of interleukin-2 (PDB ID 1M49).

Figure 7

2D and 3D interaction diagrams of (a) myricetin in the active site of mitogen-activated protein kinase 14 (PDB ID 6HWU) (b) myricetin in the active site of proto-oncogene c-Fos (PDB ID 1FOS).