Natural Product Isoliquiritigenin Activates GABA_B Receptors to Decrease Voltage-Gate Ca²⁺ Channels and Glutamate...

Reduction in glutamate release is a key mechanism for neuroprotection and we investigated the effect of isoliquiritigenin (ISL), an active ingredient of Glycyrrhiza with neuroprotective activities, on glutamate release in rat cerebrocortical nerve terminals (synaptosomes). ISL produced a concentration-dependent inhibition of glutamate release and reduced the intraterminal [Ca²⁺] increase. The inhibition of glutamate release by ISL was prevented after removing extracellular Ca²⁺ or blocking P/Q-type Ca²⁺ channels. This inhibition was mediated through the γ-aminobutyric acid type B (GABA_B) receptors because ISL was unable to inhibit glutamate release in the presence of baclofen (an GABA_B agonist) or CGP3548 (an GABA_B antagonist) and docking data revealed that ISL interacted with GABA_B receptors. Furthermore, the ISL inhibition of glutamate release was abolished through the inhibition of G_i/o-mediated responses or G_βγ subunits, but not by 8-bromoadenosine 3',5'-cyclic monophosphate or adenylate cyclase inhibition. The ISL inhibition of glutamate release was also abolished through the inhibition of protein kinase C (PKC), and ISL decreased the phosphorylation of PKC. Thus, we inferred that ISL, through GABA_B receptor activation and G_βγ-coupled inhibition of P/Q-type Ca²⁺ channels, suppressed the PKC phosphorylation to cause a decrease in evoked glutamate release at rat cerebrocortical nerve terminals.