Paeoniflorin inhibits the histaminergic system and promotes non-rapid eye movement sleep via adenosi

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Insomnia is a common disorder seen in nearly 30%-35% of the general population worldwide,with a range of 10%-15% being assessed as moderate to severe disorders.Symptoms of insomnia include poor sleep quality,difficulty in falling asleep,frequent awakenings during the night and early morning awakenings.Insomnia give rise to fatigue,irritability,impaired concentration and reduced alertness,causing serious impact on the quality of life of affected individuals.Current pharmacological treatment of insomnia involves the use of sedative-hypnotic benzodiazepines(BZ) and non-benzodiazepine drugs(Z-drugs).Z-drugs are the first line of management followed by BZ,amitryptiline and antihistamines.Insomnia could be effectively treated by the Z-drugs,such as zolpidem and the BZ,but their multiple adverse effects hamper their application.Paeoniflorin {PF,5beta-[(Benzoyloxy)methyl]tetrahydro-5-hydroxy-2-methyl-2,5-methano-1H-3,4-dioxacyclobutapentalen-1alpha(2H)-yl-beta-D-glucopyranoside,C23H28O11} is one of the principal active ingredients of Paeonia Radix.PF possesses inhibitory effects in the nervous system,such as suppress spike discharges and analgesic effect.These observations indicate that PF might have a potent sleep promoting effect.The aim of this study was to determine whether PF could modulate sleep behaviors by recording EEG and electromyogram in mice.The results showed that PF 25 and 50 mg·kg-1 could significantly shorten the sleep latency;increase the amount of non-rapid eye movement(non-REM,NREM) for 3 h,with an increase in the number of NREM and REM sleep episodes.PF 25 and 50 mg·kg-1 increased the number of bouts of wakefulness but decreased their duration.On the other hand,PF increased the number of state transitions from wakefulness to NREM sleep and subsequently from NREM sleep to wakefulness.However,PF had no effect on the amount of REM sleep.Immunohistochemical study showed that PF increased c-Fos expression in the neurons of ventrolateral preoptic area(VLPO),a sleep center in the anterior hypothalamus,and decreased c-Fos expression in the arousal tuberomammillary nucleus(TMN),which was located in the caudolateral hypothalamus.The sleep-promoting effects and changes in c-fos induced by PF were reversed by CPT,an antagonist at the adenosine A1R.We found a high degree of colocalization between histaminergic and gabaergic neurons in GAD 67-GFP mice TMN.Then we use GAD 67-GFP mice to help indentify histaminergic neurons.PF inhibited histaminergic neurons in the TMN by whole-cell patch clamping in the brain slice,CPT totally blocked the histaminergic neurons inactivation induced by PF.These results indicate that PF increased NREM and REM sleep by inhibiting the histaminergic system via A1R,suggesting its potential application for the treatment of insomnia. Insomnia is a common disorder seen in nearly 30% -35% of the general population worldwide, with a range of 10% -15% being assessed as moderate to severe disorders. Symptoms of insomnia include poor sleep quality, difficulty in falling asleep, frequent awakenings during the night and early morning awakenings .Insomnia give rise to fatigue, irritability, impaired concentration and reduced alertness, causing serious impact on the quality of life of affected individuals.Current pharmacological treatment of insomnia involves the use of sedative-hypnotic benzodiazepines (BZ ) and non-benzodiazepine drugs (Z-drugs). Z-drugs are the first line of management followed by BZ, amitryptiline and antihistamines. Insomnia could be effectively treated by the Z-drugs, such as zolpidem and the BZ, but their multiple adverse effects hamper their application. Paeoniflorin {PF, 5beta - [(Benzoyloxy) methyl] tetrahydro-5-hydroxy-2-methyl-2,5-methano-1H- 3,4-dioxacyclobutapentalen-1alpha -D-glucopyranoside, C23H28O11} is o ne of the principal active ingredients of Paeonia Radix. PF possesses inhibitory effects in the nervous system, such as suppress spike discharges and analgesic effect. The observations that that might have a potent sleep promoting effect. A purpose of this study was to determine whether PF could modulate sleep behaviors by recording EEG and electromyogram in mice. The results showed that PF 25 and 50 mg · kg-1 could significantly shorten the sleep latency; increase the amount of non-rapid eye movement (non-REM, NREM) for 3 h, with an increase in the number of NREM and REM sleep episodes. PF 25 and 50 mg · kg-1 increased the number of bouts of wakefulness but decreased their duration. On the other hand, PF increased the number of state transitions from wakefulness to NREM sleep and subsequently from NREM sleep to wakefulness. However, PF had no effect on the amount of REM sleep. Immunohistochemical study showed that PF increased c-Fos expression in the neurons of ventrolateral preoptic area (VLPO) asleep center in the anterior hypothalamus, and decreased c-Fos expression in the arousal tuberomammillary nucleus (TMN), which was located in the caudallateral hypothalamus. The sleep-promoting effects and changes in c-fos induced by PF were reviewed by CPT, an antagonist at the adenosine A1R. We found a high degree of colocalization between histaminergic and gabaergic neurons in GAD 67-GFP mice TMN. Then we use GAD 67-GFP mice to help indentify histaminergic neurons. PF are inhibited histaminergic neurons in the TMN by whole- cell patch clamping in the brain slice, CPT totally blocked the histaminergic neurons inactivation induced by PF. this results in that PF increased NREM and REM sleep by inhibiting the histaminergic system via A1R, suggesting its potential application for the treatment of insomnia.
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