南五灵安神复方改善睡眠作用的研究

Study on the sleep improvement effect of Nanwuling Anshen compound

目的研究南五灵安神复方改善睡眠的作用,并探讨其可能的起效机制。方法研究通过直接睡眠实验、对小鼠自主活动影响实验、延长戊巴比妥钠睡眠时间实验、戊巴比妥钠阈下剂量催眠实验对南五灵安神复方改善睡眠作用的功效进行验证,考察其量效关系;运用对氯苯丙氨酸(PCPA)所致失眠小鼠模型,考察南五灵安神复方改善失眠动物模型睡眠的作用;采用酶联免疫吸附测定(ELISA)试剂盒测试小鼠外周血中谷氨酸(Glu)和γ-氨基丁酸(GABA)的水平,考察南五灵安神复方改善睡眠可能的起效机制。结果自发活动实验结果表明,提取物小、中、大剂量组小鼠活动次数、站立次数、合并次数与溶剂对照组比较,差异无统计学意义(P>0.05)。提取物小、中、大剂量组小鼠外周血中Glu均低于溶剂对照组,提取物中、大剂量组小鼠外周血中GABA均高于溶剂对照组,差异具有统计学意义(P<0.05或<0.01)。实验采用的南五灵安神复方提取物小、中、大剂量组均无使小鼠直接睡眠的作用,说明南五灵安神复方在实验剂量下没有使小鼠直接睡眠的中枢抑制作用。对于戊巴比妥钠(阈上剂量)诱导的小鼠睡眠实验,提取物中、大剂量组小鼠睡眠潜伏期均短于溶剂对照组,睡眠时间均长于溶剂对照组,差异具有统计学意义(P<0.05或<0.01)。戊巴比妥钠阈下剂量催眠实验结果表明,提取物中、大剂量小鼠入睡率大于溶剂对照组,睡眠潜伏期短于溶剂对照组,差异具有统计学意义(P<0.05或<0.01)。模型对照组小鼠海马组织中GABA和5-羟色胺(5-HT)低于溶剂对照组,差异具有统计学意义(P<0.01);提取物中剂量组小鼠海马组织中GABA(32.94±5.76)ng/g和5-HT(190.15±35.27)ng/g及提取物大剂量组小鼠海马组织中GABA(35.33±7.63)ng/g和5-HT(227.42±43.94)ng/g均高于模型对照组,差异具有统计学意义(P<0.01)。结论南五灵安神复方提取物具有改善睡眠作用的功效,且具有量效关系;研究考察南五灵安神复方改善睡眠的可能起效机制为通过提高抑制性神经递质GABA浓度,降低兴奋性神经递质Glu浓度,通过提高GABA/Glu的比率起到改善睡眠的安神作用。

Objective To study the sleep improvement effect of the Nanwuling Anshen compound on improving sleep and explore its possible mechanism.Methods The effect of the Nanwuling Anshen compound on improving sleep was verified by direct sleep experiment,influence experiment on autonomous activity of mice,prolonging sleep time of pentobarbital sodium and subthreshold dose hypnosis experiment of pentobarbital sodium.Mouse model of insomnia caused by p-chlorophenylalanine(PCPA)was used to investigate the effect of Nanwuling Anshen compound in improving sleep in the animal model of insomnia;an enzyme-linked immunosorbent assay(ELISA)kit was used to test the levels of glutamate(Glu)andγ-aminobutyric acid(GABA)in the peripheral blood of mice to investigate the possible mechanism of Nanwuling Anshen compound in sleep improvement effect.Results The results of the spontaneous activity experiments showed that the differences in the number of activities,the number of standing,and the number of merging of the mice in the small,medium,and large dose groups of the extracts were not statistically significant when compared with those of the solvent control group(P>0.05).Glu in the peripheral blood of mice in the small,medium and large dose groups of the extract was lower than that of the solvent control group,and GABA in the peripheral blood of mice in the medium and large dose groups of the extract was higher than that of the solvent control group.The differences were statistically significant(P<0.05 or<0.01).The small,medium and large dose groups of Nanwuling Anshen compound extracts used in the experiment did not cause direct sleep in mice,indicating that Nanwuling Anshen compound had no effect on central inhibition of direct sleep of mice in experimental dose.For the sleep experiments induced by sodium pentobarbital(suprathreshold dose),the sleep latency of mice in the medium and large dose groups of the extract was shorter than that of the solvent control group,and the sleep time was longer than that of the solvent control group.The differences were statistically significant(P<0.05 or<0.01).The results of subthreshold dose hypnosis experiment of pentobarbital sodium showed that the sleep rate of mice in the medium and large dose groups of the extract was greater than that of the solvent control group,and the sleep latency was shorter than that of the solvent control group.The differences were statistically significant(P<0.05 or<0.01).GABA and 5-hydroxytryptamine(5-HT)in the hippocampal tissues of mice in the model control group were lower than those in the solvent control group,and the differences were statistically significant(P<0.01).GABA and 5-HT in the hippocampal tissues of mice in the medium dose group of the extract were(32.94±5.76)and(190.15±35.27)ng/g,and those in the large dose group of the extract were(35.33±7.63)and(227.42±43.94)ng/g,which were higher than those of the model control group,and the differences were statistical significance(P<0.01).Conclusion Nanwuling Anshen compound extract has the effect of improving sleep,and has the dose-effect relationship.The possible mechanism of Nanwuling Anshen compound to improve sleep is to increase the concentration of inhibitory neurotransmitter GABA and decrease the concentration of excitatory neurotransmitter Glu,and increase the ratio of GABA/Glu.