Purification And Identification?sleep-activity Evaluation?mechanism And Application Research Of Delta-sleep Inducing Peptide In Dairy Products

It is different from the traditional Chinese herbal medicine to promote sleep,which is derived from casein hydrolyzate.In this study,the natural active research engineering center of Guangdong Province has been used in the research of natural active separation,structure identification,activity evaluation,mechanism of action The field of deep foundation.The experiment was carried out from the factory(Guangzhou Green Crop Biotechnology Co.,Ltd.)to produce the casein phosphate peptide after the milk extract of the milk to obtain the slow extraction of milk peptides,with the separation and purification technology and electrophoretic patch clamp method to get milk to promote sleep peptide P3 And the promotion of sleep peptide monomer 1,monomer 2,monome r 3,monomer 4,and then through the rat pentobarbital sodium synergistic model,the study found that P3 and its sleep products not only have to shorten the time to bed,Rat sleep time,and combined with rat PCPA insomnia model,the study of milk to promote sleep apnea sleep mechanism.The structure and identification of monomer 1,monomer 2,monomer 3 and monomer 4 were explored by LTQ and electrophoretic patch clamp technique.At the same time,the milk to promote sleep peptide combined with lily,jujube kernel medicine homologous C hinese herbal medicine,combined into a better sleep effect of promoting sleep products.(1)Isolation,purification and structural identification of milk extractThe purified crude extract of milk extract was isolated and purified by medium and high pressure column chromatography,preparation of liquid phase and high efficiency analytical liquid phase separation and purification method.And then combined with LTQ-Orbitrap liquid-mass spectrometry for the identification of isolated milk-stimulating sleep peptides.The results showed that there were four peptide monomers with good sleep-promoting effect,namely monomer 1,monomer 2,monomer 3 and monomer 4,the amino acid sequences were: HQGLPQEVLN ENLLR,YKVPQLEIVPNSAEER,HPIKHQGLPQEVLN ENLLR And VPQ LEIVPNSAEER.The four monomers are the first to find a better sleep effect of the milk polypeptide.(2)Application of Electrophysical Patch C lamp Technique to Study the In Vitro Promoting Sleep Activity of BovineIn this study,we used the electrophoretic patch-clamp technique to explore the effect of milk-induced sleep peptide on the discharge of dorsal rectal neurons in the hypothalamus of rats.The The results showed that the activity of sleep-resistant peptide 1,monomer 2,monomer 3 and monomer 4 had better anti-sleep activity,and all of them had better inhibition of the average number of AP induced,and the effect of Vm was significant Change of sex.Three inhibition of VLPO neuron effect found that the best inhibition of monomer 2,while the monomer 4 to promote the effect of sleep followed by monomer 1 and 3 inhibition effect is similar.(3)Experimental study of milk-induced sleep peptides combined with pentobarbital sodiumIn this study,we used pentobarbital sodium to promote the sleep s ubstance model,and explored the effect of milk-induced sleep peptides on sleep behavior and the changes of plasma and brain neurotransmitters in rats.The results showed that compared with the normal control group,the milk leaching of sleep peptides significantly shortened the time of rats' sleep,and significantly prolonged the sleep time of rats.The time of sleep was shortened by 61.41% and sleep time was 254.20%.Pentobarbital sodium subthreshold sleep dose test,indicating that milk to promote sleep peptides can increase the sleep within 30 min ratio,compared with the normal control group,milk to promote sleep peptide group sleep rate increased by 12.50%.The results showed that in the brain tissue of the rat,in the hypothalamus,the milk stimulated sleep peptide increased the level of 5-HIAA by decreasing the 5-HT level of the awakening substance and accelerating the 5-HT metabolic rate;accelerating the metabolism of DA Sleep effect,but no effect on the monoamine neurotransmitter NE,and the results of the rat brain stem and cerebral cortex also show that milk to promote sleep peptide by regulating the level of neurotransmitters play a role in promoting sleep.(4)Effect of PCPA on Insomnia Model in RatsPCPA-induced insomnia model is a common method for the establishment of insomnia in rats.In this study,PCNP model was used to induce rat milk to promote sleep peptide.The mental state and plasma and brain neurotransmitters were observed.Variety.In the plasma of rats,it was found that the levels of N E,5-HT and DA in the plasma of the milk-stimulated peptides group were significantly different from those of the model group,indicating that the milk-induced sleep peptide could help balance the plasma Amine neurotransmitters.In the hypothalamus,the 5-HIAA / 5-HT ratio was significantly increased by increasing the 5-HT metabolism of the awakening substance,the DOPAC / DA ratio was significantly increased,the DA metabolism was accelerated,and the milk Promoting sleep apnea significantly reduced the level of awakening substance NE in the hypothalamus of rats to play a role in promoting sleep.In the cerebral cortex,the contents of 5-HT and its metabolites 5-HIAA and DA in the cerebral cortex of the rats were significantly higher than those in the model group(P <0.05),but the 5-HIAA / 5-HT ratio The DOPAC / DA ratio did not change significantly,and the DOPAC / DA ratio had no effect.Compared with the model group,the contents of 5-HT and its metabolites 5-HIAA,DA and DOPAC in the brainstem were significantly lower than those in the model group,and the awakening substance N E was significantly decreased.(5)The application of Delta-Sleep Inducing Peptide in Dairy ProductsSleep disorders have become a huge threat to people's health risks.Therefore,to find a natural auxiliary sleep raw materials,insomnia for the crowd is particularly important.In this study,the prophylaxis of warm-hearted sleep-resistant peptides combined with Ziziphus jujuba and lily was combined into a sleep-promoting oral solution,which proved that the sleep-promoting oral liquid had better effect of promoting sleep.Milk to promote sleep peptide and lily Ziziphus jujuba complex,can significantly shorten the time to sleep in rats,rats sleep time reduced by 43.47%.The sleep gro up was able to significantly prolong the sleep time of the rats,with a prolongation rate of 40.90%.Bovine sleep apnea can increase the sleep rate of rats within 30 min,compared with the normal control group,the sleep rate increased by 12.50%.In rat plasma,sleep-promoting products reduce the levels of 5-HT and DA in the plasma,and play a role in promoting sleep,without affecting NE levels in plasma.In the hypothalamus,the promotion of sleep products is mainly to reduce 5-HT,improve its metabolite 5-HIAA levels,speed up its metabolic rate,reduce hypothalamic DA levels,improve DOAPC levels,play a role in promoting sleep;does not affect rat hypothalamus NE level.In the cerebral cortex,promote sleep products by improving the level of 5-HT metabolites,improve the level of DA and its metabolites DOPAC levels,play a role in promoting sleep does not affect the level of NE rats.In the brainstem,the sleep-promoting products increased the level of 5-HIAA,accelerated the rate of 5-HT metabolism,increased the level of DOPAC,accelerated the metabolic rate of DA,decreased NE content in rat brain stem,and played the role of promoting sleep.