The Study On Sinisan Improving The Effect And Mechanism Of Sleep Based On Drosophila As Mode Organism

As a basic behavior and an important physiological phenomenon inhuman,sleep has an important relationship with social behavior,dietarybehavior and sexuality.Sleep quality has an important influence on people'slives and health.Scientists have confirmed that not only sleep can maintainthe vitality of the body,but also improve the capacity of information storageand processing by dreaming.For this reason,sleep studies are concerned moreand more by people.However,along with the rapid rhythm of modern socialliving,the exaggeration of the work pressure.Insomnia has been a commondisease and some other diseases' reason.The adjusting process of sleep is very complicated.Since sleep problemsappeared,People have been making efforts to the study on the mechanism allthe time.Recently,the establishment of models for study of human diseaseusing drosophila melanogaster has been becoming the new research lumin-escent spot.Many features of sleep in drosophila are similar to human sleep,such as behavioral,pharmacological,molecular,and genetic approaches.So,drosophila with well-established genetics had become an ideal modelorganism for sleep research.In our studies,we used an Infrared Drosophila Activity Monitor System(DAMS;TrikineticsInc.USA) to record sleep state of drosophila.Weobse-rved and analyzed the recording data for continuous 24 hr,and defined sleeptime that drosophila's quiescing time is exceed 5 min.We investigated howcan sex and age affect total sleep time and sleep-wake cycle in drosophilawith index of sleep time by analysis of data.Then we studied the dose-effectand time-effect of Freeze drying powder of sinisan in drosophila sleepimprovement by selecting the 7-day-old drosophila as observation of objects.At last,we verified the effect of Chinese medicine Freeze drying powder ofsinisan on sleep improvements in sleep-deprived Drosophila by two differentsleep deprivation ways.In the mechanism study,the first,5-HT was used theindex for the study to determine 5-HT content in the brain of drosophila, 5-HT content chang before and after the administration before and after sleepdeprivation and brain was,and then 5-HT1A receptor as an index,throughRT-PCR reaction,administration before and after sleep deprivation beforeand after the Drosophila brain 5-HT1A receptor expression differences wasstudied,from the molecular level,Drosophila freeze-dried powder ofSINISAN to improve the mechanism of sleep was studied.First,The effect of age of drosophila on the activity:the locomotoractivity of drosophila whose age is from 7 days to 17 days showed a stableand highest level.Locomotor activity of female drosophila decreasedsignificantly after 32 days,while male reduced after 22 days.In 18-38 daystime period,the activities of female Drosophila was much higher than themale Drosophila' s.Second,Our results indicated:Through observing sleep-wake cycleperiodogram,the young fruit flies have a long sleep time and a goodcontinuity.The sleep was fragmented gradually with the growth,and havesimilar features with human sleep.Third,The effect of age of drosophila on the sleep time:in 2-30 days,sleep time of male Drosophila did not significantly increase as the daysshortened.In 0-12 days,sleep time of the female Drosophila was longer,after 12 days irregular downward trend.This showed that sleep of the youngDrosophila was continuity,and with the growth of day-old sleep of adultDrosophila was gradually fragmented changes,sleep reduction.sex factors onthe impact of sleep of Drosophila was obviously,between the sleep of maleand female Drosophila was difference.In addition,under the same-day-old,sleep time of male Drosophila was significantly more than femaleDrosophila' s.Fourth,there are some difference between male and female in sleep.Female flies sleep mainly at night,but male sleep mainly at noon and night.Male flies have a longer sleep time than female. Fifth,2% drug substratum for dose,7 days for delivery time is best tomale(p＜0.01);4% drug substratum for dose,4 days for delivery time is bestto female(p＜0.01)Sixth,Freeze drying powder of sinisan has a effect onbothsleep-deprivedflies induced by light and sleep-deprived flies induced by mechanicalstimulation.Seventh,Application of ELISA detection methods,detection ofDrosophila brain 5-HT content,results showed that:7-day-old femaleDrosophila with the deepening of sleep,5-HT to maintain a certain level;andhung changes fluctuated Drosophila.Treatment group compared with theblank control group,male and female Drosophila brain 5-HT content weresignificantly different(p＜0.05);deprived of sleep deprivation anddrug-treated group,male and female Drosophila brain 5-HT content Therewere significant differences(p＜0.01);showed that freeze-dried powder ofSINISAN can promote the brain 5-HT content of Drosophila increased.Last,Application of real-time quantitative PCR method to detect theexperimental group,5-HT1A the relative expression level,results showedthat:the expression of 5-HT1A of the blank group was 4.16 times that sleepdeprivation of Drosophila so that the brain 5-HT1A lower thereby inhibitingsleep;the expression of 5-HT1A of treatment group was 3.14 times blankgroup' s that SINISAN can significantly promote sleep;At the same time,sleep deprivation treatment group of sleep deprivation was 8.88 times thegroup of sleep deprivation,so that further SINISAN can increase 5-HT1Aexpression of Drosophila brain,so as to promote sleep.