健脑丸镇静催眠作用及其机制研究

Research on the sedative-hypnotic effect and mechanism of Jiannao Wan

目的:研究健脑丸(JNW)中枢镇静催眠作用及其机制。方法:(1)MTD法研究JNW毒性。(2)KM小鼠分别随机分为JNW高、中、低剂量组、地西泮组或安神补脑液组、空白对照组,分别连续14天灌胃给予JNW、地西泮或安神补脑液及无菌蒸馏水。末次灌胃后30min,分别观察JNW对小鼠自主活动实验、旷场实验以及镇静催眠实验的影响。(3)KM小鼠分别随机分为JNW高、中、低剂量组、空白对照组,模型组,分别连续14天灌胃给予JNW及无菌蒸馏水,处死小鼠前3天,分别连续3天给予JNW高、中、低剂量组、模型组腹腔注射对氯苯丙氨酸(PCPA)建立失眠模型,末次给药后30min处死小鼠并快速冰上取脑,采用ELISA法测定海马组织中GABA、5-HT、5-HIAA、5-HT1A含量。结果:(1)JNW药物MTD为5.625 g·kg^(-1);(2)JNW低剂量组能显著减少小鼠自主活动次数(P<0.01);JNW低、中剂量组均能显著减少小鼠运动总路程(P<0.01),低、中剂量组能显著减少小鼠运动时间及平均速度(P<0.05),增加小鼠静止时间(P<0.05);JNW低、中剂量组可显著缩短小鼠睡眠潜伏期(P<0.05);JNW低、高剂量组能显著延长小鼠睡眠持续时间(P<0.01)。(3)JNW中、高剂量组可升高对氯苯丙氨酸致失眠小鼠海马组织GABA、5-HT、5-HIAA含量(P<0.05)。结论:(1)健脑丸最大耐受量为5.625 g·kg^(-1),是临床剂量的150倍。(2)健脑丸具有镇静催眠作用,其机制可能与提高海马组织GABA、5-HT、5-HIAA含量有关。

Objective: To study the central sedative-hypnotic effect and mechanism of Jiannao Wan(JNW). Method:(1The toxicity of JNW was studied by maximal tolerance dose(MTD) method.(2) KM mice were randomly divided into high-,medium-and low-dose JNW groups, diazepam or Anshen BunaoYe group, and normal control group, which were intraperitoneally administered JNW, diazepam or AnShenBuNaoYe, and sterile distilled water, respectively, for 14 consecutive days. 30 minutes after the last administration, the effects of JNW on the independent activity experiment, open field experiment, as well as sedativehypnotic experiment of mice were observed.(3) KM mice were randomly divided into high-, medium-and low-dose JNW groups,normal control group and model group, which were intraperitoneally administered JNW and sterile distilled water, respectively,for 14 consecutive days. 3 days before sacrifice, the mice of high-, medium-and low-dose JNW groups and model group were intraperitoneally injected with P-chlorophenylalanine(PCPA) for 3 consecutive days to establish an insomnia model. 30 minutes after the last administration, the mice were sacrificed and their brains were quickly removed on ice. The contents of GABA, 5-HT, 5-HIAA and 5-HT1A in the hippocampus tissue of mice were measured by ELISA. Result:(1) The MTD of JNW was 5.625 g·kg^(-1);(2) The low-dose JNW significantly reduced the number of autonomous activities of mice(P < 0.01). The low-and medium-dose JNW could significantly reduce the total running distance(P < 0.01), exercise time and average speed(P < 0.05), as well as increase the resting time of mice(P < 0.05). The low-and medium-dose JNW could significantly shorten the sleep latency of mice(P < 0.05). The lowand high-dose JNW could significantly prolong the sleep duration of mice(P < 0.01).(3) The contents of GABA, 5-HT and 5-HIAA were increased in the hippocampus of mice with PCPA-induced insomnia in the medium-and high-dose JNW groups(P<0.05).Conclusion:(1) The MTD of JNW is 5.625 g·kg^(-1), which is 150 times of the clinical dose.(2) JNW has sedative-hypnotic effects,and the mechanism is related to increasing the contents of GABA, 5-HT and 5-HIAA in hippocampus.