| Objective:The effect and mechanism of Schizandrin on improving sleeping was studied by using Drosophila as a model organism.Method:Experiment 1:We selected 2 days old flies,7 days old flies,15 days old flies, 30 days old flies,36 days old flies,40 days old flies,45 days old flies,50 days old flies as research objects (n-32), and used DAMS to monitor the activity of each fly minute-by-minute. And then used the data to assign each fly a sleep profile (sleep time and activity time as evaluation index), which included both the number of naps and total time spent asleep during the day and night, as well as the sleep-wake rhythm in different age groups.Experiment 2:According to the methods of Experiment 1, we selected 7 days old flies as research objects (n=32), and Schisandrin with 0.64mg/mL-10.16 mg/mL for 1 day as the dosage, to discuss the dose-effect relationship of the effect of Schizandrin on improving sleeping.Experiment 3:According to the methods of Experiment 1 and the optimal dosage (2.54 mg/mL for 1 day) of Experiment 2, we selected 2 days old flies, 7 days old flies,15 days old flies,30 days old flies,40 days old flies, and 45 days old flies as research objects (n=32), to discuss the effect of Schizandrin on improving sleeping and sleep-wake rhythm in different age groups.Experiment 4:According to the optimal dosage (2.54 mg/mL for 1 day) of Experiment 2, we selected 7 days old female flies, and 30 days old female flies as research objects (n=30), collected the flies'heads by using the low-temperature freezing method, and then explored the effect of Schizandrin on expression of sleep behavior related genes by using gene chip technology.Results:1.24 hours of sleep and sleep-wake rhythm in different age groups1.1 There were 24 hours of sleep in different age groups that were different. With the increase of age, female flies circadian sleep time was gradually decreased, and that of daytime naps decreased significantly. Male flies circadian sleep time was stable tends, and that of Day/Night naps were gradually decreased and then increased slightly. Starting within 30 days of age, both male and female flies Day/Night naps were slightly increased.1.2 The results shown in Sleep-Wake figure. In the low days of age (2-7 days old), Drosophila sleep-wake rhythm was not very different. With the increase of age, there was an emerging split between the sleep-wake rhythm of male and female flies. Female flies were active in daytime and sleep at night. And male flies were daytime and night sleep. Starting within 30 days of age, this phenomenon was increasingly significant.With the increase of age, female flies circadian activity time was increased, and that of daytime activity time was mainly increased, sleep tended to be more fragmented. Starting within 30 days of age, the original sleep-wake rhythm (Features:7:00am-2:00pm and 5:00-8:00pm activity,2:00-5:00pm nap, 8:00pm-6:00am sleep) was gradually replaced by the new sleep-wake rhythm (Feature:12 h-light time activity and 12h-dark time sleep).With the increase of age, male flies circadian activity time was increased, but that of activity time was scattered, sleep tended to be more fragmented. Starting within 30 days of age, the original sleep-wake rhythm (Features: 7:00am-2:00pm and 5:00-8pm activity,2:00-5:00pm sleep,8:00pm-6:00am sleep) was gradually replaced by the new sleep-wake rhythm (Feature:activity and sleep time was alternate and scattered distribution).2. The dose-effect relationship of the effect of Schizandrin on improving sleeping.2.1 Schisandrin with 0.64mg/mL-10.16mg/mL for 1 day had effect on decreasing male and female flies sleep during the daytime(p<0.01). Schisandrin with 2.54mg/mL for 1 day had significant effects on decreasing male and female flies sleep during the nighttime(p<0.05).2.2 The results shown in Sleep-Wake figure. Schizandrin had effect on sleep time, main methods to increase female fly daytime activity time and reduce fragmented sleep. And Schizandrin had effect on sleep time, main methods to increase male fly circadian activity time and reduce fragmented sleep. When the drug concentration reach 2.54mg/mL for 1 day, this phenomenon was most significant.3. The effect of Schizandrin on improving sleeping and sleep-wake rhythm in different age groups3.1 Schisandrin with 2.54mg/mL for 1 day had significant effect on decreasing different age male and female flies sleep during the nighttime(p<0.01), but had no effect on the trend of male and female flies sleep time.3.2 The results shown in Sleep-Wake figure. Compared with the control group, drug group of male and female sleep tended to be less fragmented, could recover sleep-wake rhythm to young state. Starting within 30 days of age, this phenomenon was increasingly significant.4. The effect of Schizandrin on expression of sleep behavior related genes.4.1 Compared with those of 30 days of age group, a total of 976 were differentially expressed, among which 311 were up-regulated and 665 were down-regulated in 7 days of age group.4.2 According to the optimal dosage of Experiment 2, compared with those of drug group, a total of 170 were differentially expressed, among which 86 were up-regulated and 84 were down-regulated in 7 days of age control group.4.3 According to the optimal dosage of Experiment 2, compared with those of drug group, a total of 194 were differentially expressed, among which 117 were up-regulated and 77 were down-regulated in 30 days of age control group.Conclusion:1. There are 24 hours of sleep and sleep-wake rhythm in different age groups that are different.2. Schisandrin improves male and female fruit flies sleep-wake rhythm of the optimal dosage:2.54mg/mL for 1 day.3. Schisandrin with 2.54mg/mL for 1 day has different effect on the sleep-wake rhythm of different age flies. With the increase of age, Schisandrin has effect on recovering sleep-wake rhythm to young state. 4. What 7 days old flies and 30 days old flies can do is the sleep regulation in different ways. Compared with those of 7 days of age group, the most prominent example of such genes are Or9a and Or67a up-regulated and cry down-regulated in 30 days of age group.5. Schisandrin improves the effect of 7 days of age group fruit flies sleep, is tim, per and vri major players such as genes involved in the process of multi-gene, multi-gene co-regulation results.Schisandrin improves the effect of 30 days of age group fruit flies sleep, is Gr98d and Gr93b major players such as genes involved in the process of multi-gene, multi-gene co-regulation results. |
PDF Full Text Request