Bio-affinity ultrafiltration combined with UPLC Q-Exactive Plus Orbitrap HRMS to screen potential COX-2 and 5-LOX inhibitors in mulberry (Morus alba L.) leaf

J Ethnopharmacol. 2025 Jan 4:119325. doi: 10.1016/j.jep.2025.119325. Online ahead of print.

Abstract

Ethnopharmacological relevance: Mulberry (Morus alba L.) leaf is a well-known herbal medicine in China for thousands of years. Mulberry leaf can regulate arachidonic acid (ARA) metabolism disorder in obesity and type 2 diabetes mellitus. However, the active ingredients involved in this process are still unclear.

Aim of study: To explore the potential active ingredients in mulberry leaf against ARA metabolism disorder.

Materials and methods: In this research, an efficient method combining affinity ultrafiltration, molecular docking, and network pharmacology was developed and applied to explore cyclooxygenase-2 (COX-2) and 5-lipoxygenase (5-LOX) inhibitors from mulberry leaf.

Results: A total 17 potential inhibitors were screened by affinity ultrafiltration assay and identified by high resolution mass spectrometry. In addition, 8 bioactive ingredients were obtained after re-evaluated by molecular docking and network pharmacology, and their inhibitory activities on COX-2 and 5-LOX were confirmed by in vitro inhibitory assays. The results of cell experiments showed that the expressions of COX-2 and 5-LOX were significantly suppressed by neochlorogenic acid, rutin, isoquercetin, and (-)-syringaresinol-4-O-glucoside .

Conclusion: Neochlorogenic acid, rutin, isoquercetin, and (-)-syringaresinol-4-O-glucoside may be the potential material basis of mulberry leaf in the regulation of ARA metabolism of obesity and type 2 diabetes mellitus.

Keywords: 5-lipoxygenase; affinity ultrafiltration; cyclooxygenase-2; mulberry leaf; potential inhibitor.