刺五加果水提物和醇提物对小鼠的镇静催眠作用及其机制

目的：探讨刺五加果水提物（ASF-WE）和醇提物（ASF-AE）对小鼠的镇静催眠作用，并阐明其可能作用机制，为刺五加果的开发提供依据。方法：分别制备ASF-WE和ASF-AE。120只雄性ICR小鼠随机分为空白对照组、地西泮（DZP）阳性对照组（DZP组）、不同剂量（4、8、16、32和64 mg·kg^-1） ASF-WE组和不同剂量（4、8、16、32和64 mg·kg^-1） ASF-AE组，每组10只，连续给药5 d，记录各组小鼠自主活动次数，利用阈下催眠剂量戊巴比妥钠诱导睡眠实验记录小鼠入睡率，利用催眠剂量戊巴比妥钠诱导睡眠实验记录小鼠睡眠潜伏期和睡眠时间，筛选出ASF-WE和ASF-AE的催眠剂量和亚催眠剂量。根据模型药的不同模型药分别为5-羟色氨酸（5-HTP）、对氯苯丙氨酸（pCPA）和氟马西尼（FLU）]，分别将雄性ICR小鼠40只随机分为空白对照组、模型对照组（5-HTP组和pCPA组）、ASF-WE＋模型药组和ASF-AE＋模型药组，每组10只；80只雄性ICR小鼠随机分为空白对照组、FLU组、DZP组、ASF-WE组、ASF-AE组、DZP+FLU组、ASF-WE＋FLU组和ASF-AE＋FLU组，每组10只；连续给药5 d，采用戊巴比妥钠诱导睡眠实验记录各组小鼠的睡眠潜伏期和睡眠时间，采用ELISA法检测小鼠海马组织中5-羟色胺（5-HT）和γ-氨基丁酸（GABA）水平。结果：与空白对照组比较，32和64 mg·kg^-1 ASF-WE组小鼠自主活动次数明显减少（P<0.05或P<0.01），16、32和64 mg·kg^-1ASF-AE组小鼠自主活动次数明显减少（P<0.05或P<0.01），16、32和64 mg·kg^-1ASF-WE组和ASF-AE组小鼠入睡潜伏期明显缩短（P<0.05或P<0.01），睡眠时间明显延长（P<0.05或P<0.01）。与5-HTP组比较，ASF-WE＋5-HTP组和ASF-AE＋5-HTP组小鼠睡眠潜伏期明显缩短（P<0.01），睡眠时间明显延长（P<0.01），小鼠海马组织中5-HT水平明显升高（P<0.05）。与空白对照组比较，pCPA组小鼠睡眠潜伏期明显延长（P<0.01），小鼠睡眠时间明显缩短（P<0.01），表明失眠模型成功建立；与pCPA组比较，ASF-WE＋pCPA组和ASF-AE＋pCPA组小鼠睡眠潜伏期明显缩短（P<0.05），睡眠时间明显延长（P<0.01），小鼠海马组织中GABA水平明显升高（P<0.05或P<0.01）。与ASF-WE组比较，ASF-WE+FLU组小鼠睡眠潜伏期明显延长（P<0.05），睡眠时间明显缩短（P<0.01），小鼠海马组织中GABA水平明显降低（P<0.05）；与ASF-AE组比较，ASF-AE+FLU组小鼠睡眠潜伏期差异无统计学意义（P>0.05），睡眠时间明显缩短（P<0.01），小鼠海马组织中GABA水平明显降低（P<0.05）。结论：ASF-WE和ASF-AE均具有较好的镇静催眠作用，其作用机制可能与5-HT能和GABA能中枢神经系统有关。

Sedative and hypnotic effects of water and alcohol extracts from fruits of Acanthopanax senticosus in mice and their possible mechanisms

Objective: To explore the sedative and hypnotic effects of Acanthopanax senticosus fruit water extract (ASF-WE) and alcohol extract (ASF-AE) in the mice, to elucidate the possible mechanisms, and to provide the basis for the development of Acanthopanax senticosus. Methods: ASF-WE and ASF-AE were prepared, respectively. A total of 120 male ICR mice were randomly divided into blank control group, diazepam(DZP) positive control group(DZP group), different doses (4, 8, 16, 32 and 64 mg·kg^-1) of ASF-WE groups, and different doses (4, 8, 16, 32 and 64 mg·kg^-1) of ASF-AE groups, 10 mice in each group, continuous administration for 5 d. The number of locomotor activity of the mice in various groups was recorded. The sleep rates of mice were recorded by using subthreshold dose of pentobarbital sodium induced sleep experiment. The sleep latency and sleep time of mice were recorded by using threshold dose of pentobarbital sodium induced sleep experiment. The hypnotic dose and subhypnotic dose of ASF-WE and ASF-AE were selected. According to the different model drugs, the model drugs were 5-hydroxytryptophan (5-HTP), flumazenil (FLU) and p-corophenylalanine (pCPA),40 male ICR mice were randomly divided into blank control group, model control group(5-HTP group and pCPA group), ASF-WE + model drug group and ASF-AE + model drug group, 10 mice in each group;80 male ICR mice were randomly divided into blank control group,FLU group,DZP group,ASF-WE group,ASF-AE group,DZP+FLU group,ASF-WE group and ASF-AE+FLU group,10 mice in each group, for 5 consecutive days. Pentobarbital sodium induced sleep experiment was used to record the sleep latency and sleep time of mice. ELISA was used todetect the levels of 5-hydroxytryptamine(5-HT) and gamma-aminobutyric acid (GABA) in hippocampus tissue of the mice. Results: Compared with control group, the number of locomotor activity of the mice in 32 and 64 mg·kg^-1 ASF-WE groups were significantly decreased (P<0.05 or P<0.01),and the number of locomotor activity of the mice in 16,32 and 64 mg·kg^-1ASF-AE groups were significantly decreased (P<0.05 or P<0.01). Compared with blank control group, the sleep latencies of the mice in 16,32 and 64 mg·kg^-1 ASF-WE groups were significantly reduced(P<0.05 or P<0.01),and the sleep time was significantly prolonged (P<0.05 or P<0.01); the sleep latencies of the mice in 16,32 and 64 mg·kg^-1 ASF-AE groups were significantly shortened(P<0.05 or P<0.01), and the sleep time was significantly prolonged (P<0.05 or P<0.01).Compared with 5-HTP group, the sleep latencies of the mice in ASF-WE + 5-HTP group and ASF-AE + 5-HTP group were significantly shortened (P<0.01), the sleep time was significantly prolonged (P<0.01),and the levels of 5-HT in rat hippocampus tissue were significantly increased (P<0.05). Compared with blank control group, the sleep latency of the mice in pCPA group was significantly prolonged (P<0.01), the sleep time of the mice was shortened (P<0.01), indicating that the insomnia model was successfully established. Compared with pCPA group, the sleep latencies of the mice in ASF-WE + pCPA group and ASF-AE + pCPA group were significantly shortened (P<0.05),the sleep time was significantly prolonged (P<0.01), and the GABA levels in hippocampus tissue of the mice were significantly increased (P<0.05 or P<0.01). Compared with ASF-WE group, the sleep latency of the mice in ASF-WE+FLU group was significantly prolonged (P<0.05), the sleep time was significantly shortened(P<0.01), and the GABA level in the hippocampus tissue of the mice was significantly increased (P<0.05).Compared with ASF-AE group, the difference in sleep latency in ASF-AE+FLU group was not statistically significant(P>0.05),the sleep time was significantly shortened(P<0.01), and the GABA level in the hippocampus tissue of the mice was significantly decreased(P<0.05). Conclusion: ASF-WE and ASF-AE better sedative and hypnotic effects, and their mechanism may be related to its involvement in regulating the activity of 5-HT nervous system and GABA nervous system.