Inhibitory Effects of Honokiol on Substantia Gelatinosa Neurons of the Trigeminal Subnucleus Caudalis in Juvenile Mice

Abstract

Inhibitory neurotransmitters such as gamma-aminobutyric acid (GABA) and glycine are known to be abundant in the substantia gelatinosa (SG) of the trigeminal subnucleus caudalis (Vc). Thus, it has been recognized as an initial synaptic site for regulating orofacial nociceptive stimuli. Honokiol, a principal active ingredient derived from the bark of Magnolia officinalis, has been exploited in traditional remedies with multiple biological effects, including anti-nociception on humans. However, the anti-nociceptive mechanism of honokiol on SG neurons of the Vc remains fully elusive. In this study, effects of honokiol on SG neurons of the Vc in mice were investigated using the whole-cell patch-clamp method. In a concentration-dependent manner, honokiol significantly enhanced frequencies of spontaneous postsynaptic currents (sPSCs) that were independent of action potential generation. Notably, honokiol-induced increase in the frequency of sPSCs was attributed to the release of inhibitory neurotransmitters through both glycinergic and GABAergic pre-synaptic terminals. Furthermore, higher concentration of honokiol induced inward currents that were noticeably attenuated in the presence of picrotoxin (a GABAA receptor antagonist) or strychnine (a glycine receptor antagonist). Honokiol also exhibited potentiation effect on glycine- and GABAA receptor-mediated responses. In inflammatory pain model, the increase in frequency of spontaneous firing on SG neurons induced by formalin was significantly inhibited by the application of honokiol. Altogether, these findings indicate that honokiol might directly affect SG neurons of the Vc to facilitate glycinergic and GABAergic neurotransmissions and modulate nociceptive synaptic transmission against pain. Consequently, the inhibitory effect of honokiol in the central nociceptive system contributes to orofacial pain management.