| Objective:Sleep is a basic need to humans and mammals.With the accelerated pace of life and fierce social competition,sleep deprivation(SD) has become a common and serious problem.Sleep deprivation refers to the state of sleep loss which is caused by various reasons. Sleep deprivation can not only cause changes in emotion,learning and other activities,but also has an effect on the function of the body systems and organs damages,such as the central nervous system,cardiovascular system,digestive system,respiratory system,endocrine system,immune system and reproductive system,etc. The sleep center and waking center in the mammalian brain is constituted by numerous nerve nucleis and neurotransmitters,of which the hippocampus and hypothalamus participate in the regulation of sleep-wake cycle.The imbalance caused by sleep deprivation associates with certain structure in the brain.Proteomics refers to all the proteins expressed by a cell or an organism.Essentially proteomics is to study the characteristics of protein in a mass level,including the protein expression,translation, modification and the protein interaction between proteins,so as to obtain an overall understanding of the development of disease on the protein level.We can use the related techniques of proteomics to identify the difference of protein groups under certain conditions,thus to reveals certain biological phenomena and discipline.According to proteomics studies,we can have a complete and clear understanding of the proteomics regulatory network for the occurrence and development of disease,which can provide further understand of the mechanism of the development of disease. Previous studies have demonstrated that sleep has a close relationship to the nervous degenerative disease,but the pathophysiological mechanism is not completely clear.Therefor, it is essential to understand the protein expression level on the sleep deprived condition, so as to get a comprehensive understanding of its pathophysiological mechanism.This experiment applies the protemoics technology to study the hippocampus of rats on different sleep conditions.We have isolated and identified differential proteins,in order to know the mechanism of nervous system degenerative disease caused by sleep deprivation and provide a new direction for treatment. Methods: 1 Adult male Wistar rats were divided into control group,sleep deprivation group and sleep deprivation+Vc group.Sleep deprivation group and sleep deprivation+Vc group were sleep deprived for 72 hours,while the control group kept normal wake-sleep cycle,keep the room temperature at 22 ℃and light 08:00-20:00;2 After the sleep deprivation, the rats in each group were sacrificed at the corresponding time point quickly,isolated the hippocampus on the ice tray and kept in-80 ℃condition;3 Proteins were extracted from rat hippocampus,digested by trypsin,and desalinated for next use;4 The prepared samples were tagged by TMT,buffer dissoved,strip hydration,TFA acidated and then C18 Zip-Tip desalted;5 The sample were dissolved after they were drained,and analyzed by LC-LTQ-MS/MS and searched in the database;6 The different proteins were analyzed biological function by using GeneCodis3. Results: 1 Compared to control group,a total of 99 differential proteins were identified by TMT quantitative combined with LC-MS/MS in the hippocampus of sleep deprived rats.Among these proteins,the number of up-regulated proteins is 65,while the number of down-regulated proteins is 34.Compared to sleep deprived group,a total of 20 differential proteins were identified in the hippocampus of sleep deprived+Vc group rats,the number of up-regulated proteins is 10,while the number of down-regulated proteins is 10. 2 The function of differential proteins between sleep deprivation group and control group are mainly involved in nervous system development,cell apoptosis,protein transport,protein hydrolysis,oxidative stress,low oxygen reaction long-term memory and protein ubiquitination. 3 The function of differential proteins between sleep deprivation+Vc group and sleep deprivation group are mainly involved in lipid metabolism,hydrogen peroxide metabolism,intracellular transport and protein ubiquitination. Conclusions:1 By using TMT quantitative combined with LC-MS/MS,we have identified differential proteins,built a proteome database of sleep deprived rats preliminary.The different proteins are referring to nervous system development,cell apoptosis,protein transport,protein hydrolysis,oxidative stress,low oxygen reaction,long-term memory and protein ubiquitination,and most of these processes have relationships with mechanisms of neurodegenerative diseases,especially AD.The results demonstrate that lacking of sleep may have a potential link with brain aging and neurodegenerative diseases,and can help us to get further understand of the nature of these disease. 2 Vitamin C has a protective effect in rat nervous system against the damage caused by sleep deprivation,its mechanism is mainly by reducing the oxidative stress caused by sleep deprivation. |
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