To Explore The Effects Of Schisandra Lignan On Arousal In Treadmill Sleep Deprived Rats

Objective:Patients with insomnia are in a state of 24 hours of hyperarousal,which may be the main cause of sleep perception errors and impaired daytime function in patients with insomnia.Previous studies have shown that the decrease of low-frequency delta wave power(0.5～4 Hz)during NREM sleep in insomnia patients is an important manifestation of cortical hyperarousal in insomnia patients.The previous results of our research group showed that the Schisandra lignans could antagonize the sleep time reduction in treadmill sleep deprived rats,and significantly improve the daytime function decline caused by sleep deprivation.Therefore,in this study,the low-frequency EEG power spectrum in NREM sleep period was first used as an index of arousal level to determine the effects of treadmill sleep deprivation and Schisandra lignans on the arousal level of rats.Then,based on the excitability of awakening-related nucleus neurons,we explored the mechanism of the Schisandra lignans on arousal in treadmill sleep-deprived rats.Methods:Part I: Effects of pentaphylans on EEG spectrum in treadmill sleep deprived ratsMale SD rats implanted with cortical EEG electrodes and EMG electrodes were randomly divided into model group and Schisandra lignan group.Rats in both groups were subjected to treadmill sleep deprivation for 3 days(running for 4 s and stopping for 12 s at a speed of 0.05 m/s).During sleep deprivation,rats in model group were given an equal volume of 1% CMC-Na solution,and rats in Schisandra lignans group were given Schisandra lignans suspension(67.2 mg/kg.bid)for 3 days.Using Sleep Analysis software,24 h was divided into 6 interperiods,one interperiod every 4 hours.The changes of delta wave relative power spectral density in NREM sleep period before sleep deprivation and on the 3rd day of sleep deprivation were analyzed and compared between the two groups.Part II: The effects of five seed lignans on c-Fos expression in wakefulness nuclei in treadmill sleep deprived ratsMale SD rats were randomly divided into blank group,model group,diazepam group and Schisandra lignan group.During sleep deprivation,blank group and model group were given equal volume of 1% CMC-Na solution,diazepam group was given diazepam suspension(18 mg/kg),Schisandra lignans was given Schisandra lignans suspension(67.2 mg/kg.bid)for 3 consecutive days.The brain tissues of rats in each group were collected,and the changes of c-Fos protein expression in the LH,DR,PPT,LDT and LC regions of the awakening nuclei of rats were analyzed and compared by immunofluorescence technique.Part III: The effect of five seed lignans on the action potential of DR Neurons in treadmill sleep deprived ratsMale SD rats implanted with in-vivo multichannel electrodes,were randomly divided into blank group,model group,diazepam group and Schisandra lignan group.Rats in blank group and model group were given equal volume of 1% CMC-Na solution,diazepam group was given diazepam suspension(18 mg/kg),Schisandra lignan group were given Schisandra lignan suspension(67.2 mg/kg.bid).Using in-vivo multichannel neurophysiological recording technique,the difference between neuron discharge frequency after sleep deprivation and neuron discharge frequency before sleep deprivation,ΔNeuron was used as assessment index to analyze and compare the changes of AP of neurons in dorsal raphe nucleus of rats in each group.Results:Part I: Effects of pentaphylans on EEG spectrum in treadmill sleep deprived ratsCompared with before sleep deprivation,the relative power spectral density of delta wave in the model group decreased significantly in each interphase of NREM sleep on the 3rd day of sleep deprivation(P<0.05),and the decline was the largest between 04:00-08:00.Compared with before sleep deprivation,there was no significant difference in the delta wave relative power spectral density of Schisandra lignan group during the NREM sleep period on the 3rd day of sleep deprivation(P>0.05).Compared with the model group on the 3rd day of sleep deprivation,the delta wave relative power spectral density of Schisandra lignan group in each interphase of NREM sleep was significantly increased(P<0.05),and the increase was more obvious in three intervals: 20:00-24:00,2400-04:00 and 04:00-08:00.Part II: The effects of five seed lignans on c-Fos expression in wakefulness nuclei in treadmill sleep deprived ratsCompared with blank group,c-Fos protein expression of LH,DR,PPT and LC region neurons in model group was significantly increased(P<0.01),there were no significant changes in c-Fos protein expression in LDT neurons(P>0.05).Compared with model group,the expression of c-Fos protein in LH and PPT neurons was significantly decreased in diazepam group(P<0.01),the expression of c-Fos protein in LH,DR,PPT and LC region neurons in Schisandra lignan group decreased(P<0.01).Compared with diazepam group,the expression of c-Fos protein in DR region neurons of rats in Schisandra lignan group was decreased(P<0.01).Part III: The effect of five seed lignans on the action potential of DR Neurons in treadmill sleep deprived ratsRat DR identifies two types of Neuron DR-5-HT and Neuron DR-others.Compared with blank group,ΔNeuron DR-5-HT in model group was significantly increased(P<0.01),the increase rate was more than 2 times,while the ΔNeuron DR-others changes are not significan(P>0.05).Compared with model group,there were no significant changes in diazepam group(P>0.05),ΔNeuron DR-5-HT in Schisandra lignan group decreased significantly(P<0.01),Conclusion: Schisandra lignan can reduce the arousal level of treadmill sleep deprived rats by inhibiting the activity of LH,DR,PPT and LC region neurons and the excitability of 5-HT neurons in DR area.