Based On5-ht Nervous System To Investigate The Mechanism Of Sns Active Priciples In Promoting Sleep

Clinical practice and basic experiments showed that traditional compound Chinese medicine SNS has significant improvements in sleep, but the mechanism in improving sleep is not clear. Earlier studies about the mechanism of SNS almost based on multiple-targets in the organs or nervous systems. There is no in-depth study about it. The present research based on5-HTergic nervous system to investigate the mechanism of sleep improvement of SNS. In the first section, the experiment began with sodium pentobarbital collaborative experiment, which was to definite the suprathreshold dose and the subthreshold dose in mice. Then, compared the potency of both the combined active priciples and single active priciple. After that the impact of active priciples of SNS combined with the subthreshold dose sodium pentobarbital on the sleep of mice was studied. In the second section, sodium pentobarbital collaborative experiment, enzyme linked immunosorbent assay and RT-PCR were used to clarify the mechanism of SNS in improving sleep. The present study from multiple perspectives(synthetic and metabolic of5-HT,5-HT transmitter and5-HT receptor) to investigate the mechanism of SNS.The results:1. determine threshold dose of sodium pentobarbital in mice. In the established three dose(55mg/kg,50mg/kg and45mg/kg, i.g.,4days), the suprathreshold dose of sodium pentobarbital was50mg/kg. Injecting with this dose the righting reflex of all the mice was abolition. In the established four dose(45mg/kg,40mg/kg,35mg/kg and30mg/kg, i.g.,4days), the subthreshold dose of sodium pentobarbital was35mg/kg. Injecting with this dose the righting reflex of all the mice was not abolition.2. Compared the potency of both the combined active priciples and single active priciple.Compared with vehicle group, the sleep latence of mice in the combined active priciples group, synephrine group enoxolone group and promethazine group significantly shorted (P<0.05), while sleep duration of mice in the4groups increased obviously (P<0.05). Combined active priciples group and promethazine group showed the best potency. Additionally, solvent group showed no significance either in sleep latence nor in sleep duration (P>0.05)3. Impact of subthreshold doses of sodium pentobarbital combined with SNS active priciples and single active priciple on sleep in mice.The results indicated that after injecting subthreshold doses of sodium pentobarbital, mice in vehicle group showed no righting reflex abolition, while almost all the mice in promethazine group showed righting reflex abolition. The proportion of righting reflex abolition in SNS active priciples group was66.7%.4. Impact of SNS active priciples on concentration of5-HT in the brain of mice.Compared with vehicle group, the concentration of5-HT in cortex of frontal lobe and hypothalamus of mice in SNS active priciples group increased obviously (P<0.05). however, there was a tendency to increase, the concentration of5-HT in hippocampus did not show significance (P>0.05)5. Impact of SNS active priciples combined with5-HTP on sleep latency and sleep duration.However mice in Subthreshold doses of SNS active priciples group and5-HTP group showed the tendency to increase in sleep duration and to decrease in sleep latency, there is no significance (P>0.05). when SNS active priciples group combined application of5-HT precursor5-HTP can significantly prolong sodium pentobarbital-induced sleep duration (P<0.05) and short the sleep latency (P<0.05) in mice, which showing a certain synergy.6. Impact of SNS active priciples on concentration of TPH in the brain of mice.Compared with vehicle group, the concentration of TPH in cortex of frontal lobe, hippocampus and hypothalamus of mice in SNS active priciples group increased obviously (P<0.05)7. Impact of SNS active priciples combined with PCPA on sleep latency and sleep duration.Compared with vehicle group, mice in SNS active priciples group showed shorter sleep latency (P<0.05). There was no significance among other groups (P>0.05)Compared with vehicle group, mice in SNS active priciples group showed longer sleep duration (P<0.05) while mice in PCPA group showed shorter sleep duration (P<0.05). The potency of SNS prolongs sodium pentobarbital-induced sleep time in mice can interdiction by PCPA(compared with SNS active priciples group, P<0.05). The sleep duration of mice in SNS active priciples combined PCPA group approach to the vehicle group.8. Impact of SNS active priciples on the concentration of5-HIAA and the ratio of5-HIAA/5-HT in the brain of mice.Compared with vehicle group, the concentration of5-HIAA in cortex of frontal lobe and hypothalamus of mice in SNS active priciples group increased obviously (P<0.05). however, there was a tendency to increase, the concentration of5-HIAA in hippocampus did not show significance (P>0.05)Also compared with vehicle group, the ratio of5-HIAA/5-HT in cortex of frontal lobe and hypothalamus of mice in SNS active priciples group increased obviously (P<0.05). however, there was a tendency to increase, the ratio of5-HIAA/5-HT in hippocampus did not show significance (P>0.05)9. Impact of SNS active priciples on the gene of Slc6a4and Htrla.Compared with vehicle group, the expression of Htr1a gene markedly increased while the expression of Slc6a4gene significantly decreased of mice in SNS active priciples group.Summary:Based on the above results that SNS active priciples improves sleep through increasing5-HT in various of regions of the brain. SNS active priciples can affect the synthesis, rate-limiting enzyme, metabolic, transmitter and receptor of5-HT in the brain.