Reduced c-Fos expression in orexin neurons of the lateral hypothalamic area and the locus coeruleus following injection of spinosin into mice

Spinosin, a major component of Samen Ziziphi spinosae, has been shown to modulate sedation and hypnosis; however, the underlying neuronal mechanisms of its stimulatory effects remain unclear.In the present study, we injected spinosin (15 mg/kg) or saline into mice, which were killed after 90 min. We isolated the brains, which were immunohistochemically stained for c-Fos as a biomarker for neuronal activation and assessed the expression profile of c-Fos in various sleep-arousal brain areas.Our findings revealed that there were no statistically significant differences in the expression of c-Fos in the nucleus accumbens and ventrolateral preoptic area, the vertical limb of the diagonal band nucleus, horizontal limb of the diagonal band nucleus, ventral tuberomammillary nucleus, ventral tegmental area, and dorsal raphe nucleus relative to saline between saline and spinosin-treated mice. Unlike saline, spinosin markedly decreased c-Fos expression in the lateral hypothalamic area (LHA) as well as the locus coeruleus (LC). Compared to the saline injection, the application of spinosin also resulted in a marked decrease in c-Fos expression in the LHA orexin neurons.These findings suggest that spinosin administration results in a restricted pattern of c-Fos expression within the LHA orexin neurons and the LC, suggesting that this particular neuronal inactivation contributes to sedation and hypnosis.