Plasma metabolomics and network pharmacology analyses combined identify the action of Xiongzhi Dilong decoction in migraine treatment

Abstract

Objective

To elucidate the mechanism of action of Xiongzhi Dilong decoction (XZDLD) in calcitonin gene-related peptide (CGRP)-induced migraine using network pharmacology and plasma metabolomic analyses.

Methods

Ultra-high-performance liquid chromatography-linear trap quadrupole orbitrap mass spectrometry (UPLC-LTQ-Orbitrap-MS) was used to identify the chemical components of XZDLD. A rat model of migraine was established by injecting a calcitonin gene-related peptide into the dura mater cannula. Migraine-like behavior was observed. Mechanical and thermal pain thresholds were measured to evaluate the efficacy of XZDLD in treating migraine. Crucial XZDLD targets in migraine were identified using network pharmacology and plasma metabolomics. Relevant proteins were validated using western blotting and immunohistochemistry.

Results

UPLC-LTQ-Orbitrap-MS was used to identify 38 compounds. Compared with the model rats, XZDLD significantly improved behavior at the first 30 min (all P < .05) and increased mechanical pain thresholds on day 19–22 (all P < .05) and thermal pain thresholds on day 20–22 (all P < .05). Moreover, network pharmacology identified 128 potential targets of XZDLD related to migraine. The metabolomic results suggested 17 differential endogenous metabolites, mainly involved in tryptophan and tyrosine metabolic pathways, in the control, model, and XZDLD groups. Six key targets were identified and used for molecular validation: MAOA, MAOB, CYP1A1, CYP1A2, CYP1B1, and ADH1C. Compared with the model group, XZDLD significantly decreased the expression of MAOA, MAOB, and CYP1B1 proteins (P = .024, P = .004, P = .039, respectively) and elevated the expression of CYP1A1, CYP1A2, and ADH1C (P = .033, P = .045, P = .027, respectively).

Conclusion

XZDLD can improve behavior and increase pain threshold. Moreover, XZDLD exerts anti-migraine effects primarily by modulating tryptophan and tyrosine metabolic pathways, providing a novel perspective on migraine research that is beneficial for its clinical application.