Integrated Phytochemical Analysis Based on UPLC–MS/MS and Network Pharmacology Approaches to Explore the Effect of Odontites vulgaris Moench on Rheumatoid Arthritis

Abstract

<i>Odontites vulgaris</i> Moench has the effect of clearing away heat, detoxification, dispelling wind, and clearing dampness. In this study, the potential anti-inflammatory compounds of <i>O. vulgaris</i> were investigated using ultra-high-performance liquid chromatography-mass spectrometry (UPLC-MS/MS) combined with the network pharmacology approach and further confirmed on an LPS-activated RAW 264.7 macrophage model. Monomer compounds were prepared from the active fraction using modern advanced separation and purification methods. UPLC-Q-Exactive HRMS was used to identify the chemical compounds in the active fractions of <i>O. vulgaris</i>. D-mannitol, geniposidic acid, salidroside, shanzhiside methyl ester, eleutheroside B, geniposide, 7,8-dihydroxycoumarin, gardoside methyl ester, arenarioside, vanillic acid, p-hydroxy-cinnamic acid, melampyroside, syringaresinol, tricin, and diosmetin were isolated from <i>O. vulgaris</i> for the first time. A compound database of <i>O. vulgaris</i> was established based on the existing literature to predict the mechanism of <i>O. vulgaris</i> in the treatment of rheumatoid arthritis. The results suggest that the PI3K-Akt pathway mediates <i>O. vulgaris</i> and deserves more attention in the treatment of RA. Finally, the anti-rheumatoid arthritis effects of the four target compounds were validated with the decreased levels of NO, TNF-<i>α</i>, IL-6 and IL-1<i>β</i> in RAW 264.7 macrophage cells treated with LPS. The present study explored the potential targets and signaling pathways of <i>O. vulgaris</i> in the treatment of RA, which may help to illustrate the mechanisms involved in the action of <i>O. vulgaris</i> and may provide a better understanding of the relationship between <i>O. vulgaris</i> and RA. This study provides novel insights into the development of new drugs and utilization of Mongolian traditional Chinese medicine resources.