| Object:Study on the mechanism of sleep-improving effect of schisandrin.Method:1. The effect of schisandrin on sleep duration and sleep-wake rhythm in 24h of 7-day old Canton S Drosophila:Picked out newly metamorphosed virgin drosophila, no older than 8h, sexually separated male and female flies under anaesthesia by CO2 and bred them in glass tubes with basic medium. Both male and female flies were then divided into blank control group and medication administration group randomly 6 days later,32 samples each group, and all samples were immigrated into monitoring tubes before 7 pm which were filled with medicated medium for medication administration group and basic medium for blank controlled group. After 12h adaptation, sleep duration and sleep-wake rhythm of drosophila in 24h from 7 am in 7th day and 7 am in 8th day was recorded by DAMS.2. The effect of schisandrin on sleep duration and sleep-wake rhythm in 24h of 7-day old Canton S Drosophila which were sleep deprived by light: Picked out newly metamorphosed virgin drosophila, no older than 8h, sexually separated male and female flies under anaesthesia by CO2 and bred them in glass tubes with basic medium. Both male and female flies were then divided into blank control group, sleep-deprived group and medication administration group (flies in this group were sleep deprived as well) randomly 6 days later,32 samples each group, and all samples were immigrated into monitoring tubes before 7 pm which were filled with medicated medium for medication administration groups and basic medium for blank controlled group and sleep-deprived group. After 12h adaptation, sleep duration and sleep-wake rhythm of drosophila in 24h from 7 am in 7th day and 7 am in 8th day was recorded by DAMS.3. Dose-effect relationship of 3-iodo-L-tyrosine on sleep duration in 24h of 7-day old Canton S Drosophila:Picked out newly metamorphosed virgin drosophila, older than 8h, sexually separated male and female flies under anaesthesia by CO2 and bred them in glass tubes with basic medium. Both male and female flies were then divided into blank control group and medication administration groups of different dosage randomly 6 days later, 32 samples each group, and all samples were immigrated into monitoring tubes before 7 pm which were filled with medicated medium of different concentration for medication administration groups and basic medium for blank controlled group. After 12h adaptation, sleep duration and sleep-wake rhythm of drosophila in 24h from 7 am in 7th day and 7 am in 8th day was recorded by DAMS.4. The effect of co-administration of schisandrin and 3-iodo-L-tyrosine on sleep duration and sleep-wake rhythm of 7-day old Canton S Drosophila: Picked out newly metamorphosed virgin drosophila, no older than 8h, sexually separated male and female flies under anaesthesia by CO2 and bred them in glass tubes with basic medium. Both male and female flies were then divided into blank control group, schisandrin administration group, 3-iodo-L-tyrosine administration group and co-administration group randomly 6 days later,32 samples each group, and all samples were immigrated into monitoring tubes before 7 pm which were filled with different medicated medium for different medication administration groups and basic medium for blank controlled group. After 12h adaptation, sleep duration and sleep-wake rhythm of drosophila in 24h from 7 am in 7th day and 7 am in 8th day was recorded by DAMS.5. Dose-effect relationship of 3,4-Dihydroxy-L-phenylalanine on sleep duration in 24h of 7-day old Canton S Drosophila:Picked out newly metamorphosed virgin drosophila, older than 8h, sexually separated male and female flies under anaesthesia by CO2 and bred them in glass tubes with basic medium. Both male and female flies were then divided into blank control group and medication administration groups of different dosage randomly 6 days later,32 samples each group, and all samples were immigrated into monitoring tubes before 7 pm which were filled with medicated medium of different concentration for medication administration groups and basic medium for blank controlled group. After 12h adaptation, sleep duration and sleep-wake rhythm of drosophila in 24h from 7 am in 7th day and 7 am in 8th day was recorded by DAMS.6. Theeffect of co-administration of schisandrin and 3,4-Dihydroxy-L-phen-ylalanine on sleep duration and sleep-wake rhythm of 7-day old Canton S Drosophila:Picked out newly metamorphosed virgin drosophila, no older than 8h, sexually separated male and female flies under anaesthesia by CO2 and bred them in glass tubes with basic medium. Both male and female flies were then divided into blank control group, schisandrin administration group,3,4-Dihydroxy-L-phenylalanine administration group and co-administration group randomly 6 days later,32 samples each group, and all samples were immigrated into monitoring tubes before 7 pm which were filled with different medicated medium for different medication administration groups and basic medium for blank controlled group. After 12h adaptation, sleep duration and sleep-wake rhythm of drosophila in 24h from 7 am in 7th day and 7 am in 8th day was recorded by DAMS.7. The effect of schisandrin on the content of dopamine in the brain of 7-day old Canton S Drosophila:Picked out newly metamorphosed virgin drosophila, no older than 8h, sexually separated male and female flies under anaesthesia by CO2 and bred them in glass tubes with basic medium. Both male and female flies were then divided into blank control group and medication administration group randomly 6 days later,50 samples each group. Frozen all the flies with liquid nitrogen and cut down heads which were later conserved in freezing tubes, each tube contained 40 heads. Heads, soaked in physiological saline, were grinded into pieces in a homogenizer, and centrifuged at 3000rpm, lOmin. Collected supernatant and determined the content of dopamine.Result:1. Schisandrin (2.54mg/ml) was able to reduce sleep duration of both male and female drosophila significantly. 2. There was no significant difference between the sleep duration of female medication administration group and female sleep-deprived group, while difference was significant between groups of male, and the sleep duration was reduced sharply. Rhythm figure of male medication administration group demonstrated that it was harder to fall asleep and easier to arouse.3. Sleep duration of 7-day old female drosophila was significantly increased by administration of 3-iodo-L-tyrosine just one day, in dosage of 0.4mg/ml～3.2mg/ml, while that of male drosophila was not increased until dosage was added up to 1.6mg/ml～3.2mg/ml.4. Sleep duration of both co-administration group 1 revealed no significant difference, comparing to SCH group and 3-iodo-group; however that of both co-administration group 2 revealed significant difference.5. Administrated with 3,4-Dihydroxy-L-phenylalanine just one day, the sleep duration of female drosophila was significantly reduced at dosage of 0.4mg/ml, and the sleep duration of male drosophila was significantly reduced at dosage of 0.2mg/ml.6. Although there was no significant difference of whole sleep duration of female co-administration group, comparing to Schisandrin group and L-dopa group, the daytime sleep duration was significantly increased while the nighttime sleep duration was significantly reduced and sleep times were increased. And all these indexes of male groups were significantly changed.7. Content of dopamine of schisandrin groups were inclined to reduce, but there was no significant difference comparing to blank controlled group.Conclusion:1. Schisandrin is able to reduce the sleep duration of 7-day old drosophila.2. Drosophila, suffered sleep deprivation caused by light, is harder to fall asleep and easier to arouse.3. The best dosage of 3-iodo-L-tyrosine to increase sleep duration of 7-day old female drosophila is 4mg/10ml, and that for male drosophila is 16mg/10ml. 4. Co-administration group 1 could not impact the effect of 3-iodo to 7-day old drosophila, to the contrast co-administration group 2 is capable of impact the effect of 3-iodo to 7-day old drosophila which proves that schisandrin may impact generation of dopamine by interfere the process of decarboxylation of dopa.5. The best dosage of 3,4-Dihydroxy-L-phenylalanine to reduce sleep duration of 7-day old female drosophila is 4mg/10ml, and that for male drosophila is 2mg/10ml.6. Schisandrin is able to reduce sleep duration of male drosophila much sharper with co-administration of L-dopa at subthreshold dose7. Schisandrin is able to inhibit generation of dopamine in brain of 7-day old drosophila. |
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