Identification of key bioactive anti-migraine constituents of Asari radix et rhizoma using network pharmacology and nitroglycerin-induced migraine rat model

Purpose: To elucidate the bioactive constituents of Asari radix et rhizoma (ARR) in treating migraine based on network pharmacology and nitroglycerin-induced migraine rat model. Methods: The potential bioactive constituents of ARR were identified with the aid of literature retrieval and virtual screening, and the migraine-related hub genes were identified using protein-protein interaction and topology analyses. Then, the interaction between the potential bioactive constituents and hub genes was determined with molecular docking and topology, leading to the prediction of the anti-migraine constituents of ARR. Moreover, a rat model of nitroglycerin-induced migraine was used to confirm the prediction by measuring the frequency of head-scratching and head-shaking behavior (FHHB) in the rats. In addition, levels of nitric oxide (NO) and calcitonin gene-related peptide (CGRP) in blood, norepinephrine (NE) and 5-hydroxytryptamine (5-HT) in brain were measured using appropriate commercial kits. Results: Network pharmacology revealed that naringenin-7-O-β-D-glucopyranoside and higenamine might be the key anti-migraine bioactive constituents of ARR. On addition of naringenin-7-O-β-D- glucopyranoside or higenamine to ARR, there was marked enhancement of the mitigating effect of ARR on nitroglycerin-induced abnormalities in levels of NO, CGRP, 5-HT and NE, as well as FHHB in rats (p < 0.05 or 0.01). Conclusion: These findings indicate that naringenin-7-O-β-D-glucopyranoside and higenamine might be the key bioactive and anti-migraine constituents of ARR. However, in addition to naringenin-7-O-β-D- glucopyranoside and higenamine, there were many other anti-migraine constituents in ARR. Therefore, there is need for further investigations on the actual contributions of these two constituents of ARR in treating migraine.