Objective To investigate the effects of Schisandrae Chinensis Fructus lignans on the alertness of the rats with sleep deprived by treadmill exercise and the underlying neurobiological mechanism. Methods According to the random number table method,SD male rats were assigned into control,sleep deprivation,low-,medium-,and high-dose Schisandrae Chinensis Fructus lignans,and atomoxetine hydrochloride groups,with 8 rats in each group.The rats in other groups except the control group were subjected to sleep deprivation by treadmill exercise for 3 d.During the deprivation period,each administration group was administrated with the corresponding drug by gavage,and a 5-9 hole tester was used to test the alertness performance of rats in each group. Furthermore,other SD male rats were selected and randomized into control,sleep deprivation,Schisandrae Chinensis Fructus lignans (67.2 mg/kg) and atomoxetine hydrochloride groups,with 10 rats in each group.The rats were modeled with the sleep deprivation method the same as that above and administrated with corresponding agents.ELISA was employed to measure the serum level of orexin A in each group of rats.The protein levels of c-Fos,orexin receptor 1,and orexin receptor 2 in the prefrontal cortex of rats in each group were observed by immunofluorescence and Western blotting. Results Compared with the control group,sleep deprivation reduced the choice accuracy (P<0.001) and increased the omission responses,omission percent,and mean correct response latency (P=0.002,P=0.003,P=0.020).Compared with the sleep deprivation group,medium- and high-dose Schisandrae Chinensis Fructus lignans and atomoxetine hydrochloride improved the alertness of rats,as demonstrated by the increased choice accuracy (P=0.001,P=0.006,P<0.001) and reduced omission responses (P=0.001,P=0.001,P<0.001),omission percent (P=0.001,P=0.002,P<0.001),and mean correct response latency (P=0.018,P=0.003,P=0.014).Compared with the control group,the sleep deprivation group showed elevated level of orexin A in the serum (P<0.001),up-regulated expression of c-Fos (P<0.001),and down-regulated expression of orexin receptor 1 (P=0.037) in the prefrontal cortex.Compared with the sleep deprivation group,Schisandrae Chinensis Fructus lignans (67.2 mg/kg) and atomoxetine hydrochloride lowered the orexin A level in the serum (P=0.005,P=0.029),down-regulated the expression of c-Fos (P=0.028,P=0.036),and up-regulated the expression of orexin receptor 1 (P=0.043,P=0.013) in the prefrontal cortex. Conclusion Schisandrae Chinensis Fructus lignans may antagonize the alertness decrease caused by sleep deprivation by regulating the secretion of orexin and the expression of orexin receptor 1 in the prefrontal cortex.
目的 探讨五味子木脂素对跑步机睡眠剥夺大鼠警觉性水平的影响及其可能的神经生物学机制。方法 SD雄性大鼠,随机数字表法分为空白组,睡眠剥夺组,五味子木脂素低、中、高剂量组,盐酸托莫西汀组,每组8只。除空白组外,其他各组大鼠均进行跑步机睡眠剥夺,连续3 d。剥夺期间,各个给药组分别灌胃给予相应药物,采用5~9孔测试仪测试各组大鼠的警觉性表现;另取SD雄性大鼠,睡眠剥夺方法造模和给药方法同上,随机数字表法分为空白组、睡眠剥夺组、五味子木脂素组(67.2 mg/kg)和盐酸托莫西汀组,每组10只,采用ELISA技术测定各组大鼠血清中食欲素A的含量;采用免疫荧光、Western blot法观察各组大鼠前额叶皮层(PFC)c-Fos蛋白表达和食欲素受体 1、2水平的变化。结果 与空白组相比,睡眠剥夺组大鼠警觉性表现的选择准确性显著降低(P<0.001),遗漏反应、遗漏率和平均反应潜伏期均显著升高(P=0.002,P=0.003,P=0.020);与睡眠剥夺组相比,五味子木脂素中、高剂量组和盐酸托莫西汀组均可改善跑步机睡眠剥夺大鼠的警觉性指标,表现为选择准确性提高(P=0.001,P=0.006,P<0.001),遗漏反应(P=0.001,P=0.001,P<0.001)、遗漏率(P=0.001,P=0.002,P<0.001)和平均反应潜伏期(P=0.018,P=0.003,P=0.014)减少。与空白组相比,睡眠剥夺组大鼠血清中的食欲素 A含量、PFC脑区c-Fos均显著增加(P均<0.001),食欲素受体1蛋白表达水平显著减少(P=0.037);与睡眠剥夺组相比,五味子木脂素组(67.2 mg/kg)和盐酸托莫西汀组大鼠血清中的食欲素 A水平(P=0.005,P=0.029)、PFC脑区c-Fos(P=0.028,P=0.036)均显著降低,食欲素受体1蛋白表达水平显著升高(P=0.043,P=0.013)。结论 五味子木脂素可能通过调节食欲素分泌和PFC的食欲素受体1表达发挥拮抗睡眠剥夺所致警觉性水平下降的作用。.
Keywords: Schisandrae Chinensis Fructus lignans; alertness; orexin; sleep deprivation.