调和肝脾方改善睡眠的时效关系及作用机制的研究

Study on the time-effect relationship and mechanism of decoction for regulating liver and spleen for improving sleep

目的研究调和肝脾方改善睡眠的时效关系及作用机制。方法实验一:将体质量20~22 g ICR雄性小鼠90只随机分为空白对照组10只、调和肝脾方中剂量组40只和调和肝脾方高剂量组40只,调和肝脾方中、高剂量组分别给予相应剂量调和肝脾方灌胃,空白对照组给予同体积蒸馏水灌胃,均每日1次。调和肝脾方中、高剂量组分别于干预3,5,7,9 d给药1 h后各随机取10只,分别腹腔注射40 mg/kg(阈剂量)的戊巴比妥钠溶液,记录小鼠的睡眠潜伏期以及睡眠时间。实验二:分组及干预方式同实验一,调和肝脾方中、高剂量组分别于干预3,5,7,9 d给药30 min后各随机取10只,分别腹腔注射30 mg/kg(阈下剂量)戊巴比妥钠溶液,计算入睡率。实验三:将体质量20~22 g ICR雄性小鼠30只随机分为空白对照组、调和肝脾方中剂量组、调和肝脾方高剂量组,每组10只。调和肝脾方中、高剂量组分别给予相应剂量调和肝脾方灌胃,空白对照组给予同体积蒸馏水灌胃,均每日1次,连续灌胃7 d,于末次灌胃30 min后,将各组小鼠直接断头处死,检测皮质、下丘脑、海马体中多巴胺(DA)、去甲肾上腺素(NE)、5-羟色胺(5-HT)、一氧化氮(NO)、一氧化氮合酶(NOS)含量。结果调和肝脾方中、高剂量组灌胃5,7,9 d的睡眠潜伏期均显著短于空白对照组(P均<0.05);调和肝脾方中剂量组灌胃7,9 d和调和肝脾方高剂量组灌胃5,7,9 d的睡眠时间均显著长于空白对照组(P均<0.05)。调和肝脾方中剂量组灌胃5,7,9 d和调和肝脾方高剂量组灌胃3,5,7,9 d的小鼠入睡率均显著高于空白对照组(P均<0.05)。调和肝脾方中剂量组海马体DA、NE含量和皮质NO、DA、NE、NOS含量均显著低于空白对照组(P均<0.05);调和肝脾方高剂量组皮质NOS、NE含量和丘脑NO含量均显著低于空白对照组(P均<0.05)。结论调和肝脾方改善睡眠的作用呈现一定的时效关系,其给药5 d起效,最佳给药时间为7 d。调和肝脾方改善睡眠的作用与调节中枢神经递质DA、NE及NO含量相关。

Objective It is to study the the time-effect relationship and mechanism of decoction for regulating liver and spleen for improving sleep(DRLSI).Methods Experiment I:90 male ICR male mice weighing 20-22g were randomly divided into blank control group(n=10),middle-dose group of DRLSI(n=40)and high-dose group of DRLSI(n=40).The middle-dose group and high-dose group were respectively given the corresponding dose of DRLSI by gavage,and the blank control group was given the same volume of distilled water by gavage,all once a day.10 mice in the middle and high dose groups were randomly selected and given pentobarbital solution 40 mg/kg(threshold dose)by intraperitoneal injection in 1 h after intervention for 3,5,7 and 9 days,the sleep latency and the time of sleep were recorded.Experiment II:The grouping and intervention methods were the same as those in experiment I.10 mice in the middle and high dose groups were randomly selected and given pentobarbital solution 30 mg/kg(subthreshold dose)by intraperitoneal injection in 30 min after intervention for 3,5,7 and 9 days,the sleep rate was calculated.Experiment III:30 male ICR male mice weighing 20-22g were randomly divided into blank control group(n=10),middle-dose group of DRLSI(n=10)and high-dose group of DRLSI(n=10).The middle-dose group and high-dose group were respectively given the corresponding dose of DRLSI by gavage,and the blank control group was given the same volume of distilled water by gavage once a day,all the mice were treated for 7 days.After the last gavage for 30 minutes,the mice in each group were directly decapitated to detect the contents of dopamine(DA),norepinephrine(NE),serotonin(5-HT),nitric oxide(NO),and nitric oxide synthase(NOS)in the cortex,hypothalamus,and hippocampus.Results The sleep latency of the mice in the middle and high dose groups was significantly shorter than that of the blank control group after gavage for 5,7,9 days(P<0.05).The sleep time of the middle dose group after gavage for 7,9 days and than in high dose group after gavage for 5,7,9 days was significantly longer than that of the blank control group(P<0.05).The rate of falling to sleep of the mice in the middle dose group after gavage for 5,7,9 days and that in the high dose group after gavage for 3,5,7,9 days was significantly higher than that of the control group(P<0.05).The content of DA and NE in hippocampus and the contents of NO,DA,NE and NOS in cortex were significantly lower in the middle dose group than those in the control group(P<0.05).The content of NOS and NE in cortex and NO in the thalamus in the high dose group was significantly lower than that in the blank control group(P<0.05).Conclusion DRLSI could improve sleep which depend on a certain time-effect relationship.It takes 5 days to take effect,and the optimal administration time is 7 days.The effect is related to the regulation of central nervous system DA,NE and NO levels.