辽宁中医药大学学报

目的 观察养血安神中药首乌藤、化痰宁心中药远志与养心安神中药炒酸枣仁对失眠模型小鼠一般情况、入睡潜伏期、睡眠维持时间、睡眠相关神经递质、激素及蛋白表达的影响，探讨不同药物安神作用强弱和作用特点，并初步探讨作用机制。方法 采用皮下注射PCPA构建失眠模型小鼠，将模型小鼠随机分为8组，分别为模型组、阳性药组、炒酸枣仁（低剂量组、高剂量组）、制远志水（低剂量组、高剂量组）和首乌藤（低剂量组、高剂量组），每组10只，另取10只小鼠作为空白对照组。阳性药组灌胃艾司唑仑混悬液，各中药组分别灌胃相应水煎剂，空白对照组和模型组灌胃等容积蒸馏水，各组小鼠灌胃容积均为20 mL/kg，连续灌胃7 d。观察小鼠一般状况、入睡潜伏期及睡眠持续时间，并用酶联免疫吸附试验（ELISA）检测小鼠脑组织神经递质及激素含量水平（5-HT、GABA、Glu、NE、MT、CRH），采用Western blotting检测Clock及BMAL1蛋白的表达。结果 与空白对照组比较，模型组小鼠易激惹、精神萎靡、皮毛枯槁蓬乱，小鼠体质量明显降低（P<0.05），入睡潜伏期显著延长、睡眠持续时间明显缩短（P<0.05），脑组织5-HT和GABA含量水平降低（P<0.05）,Glu和CRH含量水平升高（P<0.05）,Clock、BMAL1蛋白表达降低。与模型组比较，除远志高剂量组、首乌藤低剂量组外，炒酸枣仁高剂量组、炒酸枣仁低剂量组、首乌藤高剂量组、制远志低剂量组小鼠体质量均显著增加（P<0.05）；各给药组均能显著缩短小鼠入睡潜伏期（P<0.05），其中炒酸枣仁高剂量组作用最为明显；炒酸枣仁高剂量组、制远志低剂量组、首乌藤高剂量组均能显著延长小鼠睡眠持续时间（P<0.05），其中制远志组最为明显；首乌藤高剂量组、首乌藤低剂量组、酸枣仁高剂量组、酸枣仁低剂量组小鼠5-HT含量均明显增加（P<0.05），酸枣仁高剂量组小鼠脑组织GABA含量均明显增加（P<0.05），制远志高剂量组、制远志低剂量组、炒酸枣仁高剂量组、首乌藤低剂量组小鼠脑组织Glu含量均明显降低（P<0.05），首乌藤低剂量组、炒酸枣仁高剂量组、首乌藤高剂量组小鼠脑组织MT含量均明显增加（P<0.05），首乌藤低剂量组、首乌藤高剂量组、炒酸枣仁高剂量组、炒酸枣仁低剂量组小鼠脑组织CRH含量均明显降低（P<0.05），阳性药组显著增加BMAL1蛋白表达（P<0.05），制远志低剂量组明显增加Clock蛋白表达（P<0.05）。同种中药高低剂量组比较，炒酸枣仁高剂量组对缩短小鼠入睡潜伏时间明显优于低剂量组（P<0.05），制远志低剂量组较高剂量组更能显著延长小鼠睡眠持续时间（P<0.05）；各中药组小鼠5-HT、GABA、Glu、NE和MT含量以及Clock、BMAL1蛋白表达差异均无统计学意义（P>0.05）。结论 炒酸枣仁、制远志及首乌藤均有改善失眠的作用，其中高剂量炒酸枣仁在促进入睡时间方面疗效突出，制远志在维持睡眠时间方面有良好作用，炒酸枣仁、制远志、首乌藤均具有显著的助眠作用，大剂量应用可显著提高疗效并无明显不良反应；低剂量制远志在延长睡眠持续时间方面具有明显优势，同时具有一定改善睡眠节律紊乱的作用，但大剂量应用并不能增加疗效，并具有明显不良反应。安神中药可能通过促进小鼠神经递质5-HT、GABA、MT合成和分泌，抑制CRH和Glu能神经系统而发挥助眠作用。

Objective To observe the effects of the blood nourishing and tranquilizing traditional Chinese medicine Shouwuteng(Polygoni Multiflori Caulis),the phlegm transforming and tranquilizing traditional Chinese medicine Yuanzhi(Polygdae Padix),and the heart nourishing and tranquilizing traditional Chinese medicine fried Suanzaoren(Ziziphi Spinosae Semen) on the insomnia model mice's general condition,sleep latency,sleep maintenance time,sleep-related neurotransmitters,hormones,and protein expression,and to investigate the strength of tranquilizing effects of the different medicines and their effects characteristics,and to explore the mechanism of action initially. Methods The insomnia model mice were constructed by subcutaneous injection of PCPA,and the model mice were randomly divided into 8 groups,which were the model group,the positive drug group,the fried Suanzaoren(low-dose group,high-dose group),the preparation of Yuanzhi water(low-dose group,high-dose group)and Shouwuteng(low-dose group,high-dose group),with 10 mice in each group,and 10 mice were taken as the blank control group. The positive drug group was gavaged with eszopiclone suspension,each traditional Chinese medicine group was gavaged with the corresponding decoction,the blank control group and the model group were gavaged with an equal volume of distilled water,and the gavage volume of mice in each group was 20 mL/kg,and the gavage was performed continuously for 7 d. Observations on the general condition of the mice,the latency to sleep and the duration of sleep were performed,and the levels of neurotransmitters and hormones in the brain tissue of the mice were detected using enzyme-linked immunosorbent assay(ELISA)(5-HT,GABA,Glu,NE,MT,CRH),and the expression of Clock and BMAL1 proteins were detected by Western blotting. Results Compared with the blank control group,mice in the model group were irritable,depressed,withered and disheveled fur,with significantly lower body weight(P<0.05),significantly prolonged sleep latency and shortened sleep duration(P<0.05),with reduced levels of 5-HT and GABA content in the brain tissues(P<0.05),and increased levels of Glu and CRH content(P<0.05),and reduced expression of Clock and BMAL1 proteins(P<0.05). Clock and BMAL1 protein expression decreased. Compared with the model group,except for the high-dose group of Yuanzhi and the low-dose group of Shouwuteng,the body weight of mice in the high-dose group of fried Suanzaoren,the low-dose group of fried Suanzaoren,the high-dose group of Shouwuteng and the low-dose group of prepared Yuanzhi were all significantly increased(P<0.05);all the administered groups could significantly shorten the latent period of mice to fall asleep(P<0.05),with the high-dose group of fried Suanzaoren being the most obvious;the high dose group of fried Suanzaoren,the low-dose group,Shouwuteng high-dose group can significantly prolong the duration of sleep in mice(P<0.05),among which the effect of prepared Yuanzhi group is the most obvious;Shouwuteng high-dose group,Shouwuteng low-dose group,Suanzaoren highdose group,Suanzaoren low-dose group,the content of 5-HT in mice were all significantly increased(P<0.05),Suanzaoren high dose group,the GABA content of mouse brain tissue was significantly increased(P<0.05). The Glu content in the brain tissue of mice in the high dose group of prepared farnesol,low dose group of prepared farnesol,high dose group of fried Suanzaoren,and low dose group of Shouwuteng were all significantly decreased(P<0.05),the MT content in the brain tissue of mice in the low dose group of Shouwuteng,high dose group of fried Suanzaoren and high dose group of Shouwuteng were all significantly increased(P<0.05),the Glu content in the brain tissue of mice in the low dose group of Shouwuteng,the low dose group of Shouwuteng,the high dose group of Shouwuteng,the high dose group of fried Suanzaoren,and the low dose group of fried Suanzaoren low-dose group mice brain tissue CRH content were significantly reduced(P<0.05),positive drug group significantly increased BMAL1 protein expression(P<0.05),the prepared of farnesol low-dose group significantly increased Clock protein expression(P<0.05).Comparing the high and low dose groups of the same traditional Chinese medicine,the high dose group of fried Suanzaoren was significantly better than the low dose group in shortening the latency time of mice to fall asleep(P<0.05),and the low dose group of prepared Yuanzhi significantly prolonged the duration of sleep of the mice compared with the high dose group(P<0.05);the contents of 5-HT,GABA,Glu,NE,and MT,as well as the expression of Clock and BMAL1 proteins of the mice in all the traditional Chinese medicine groups did not differ significantly(P>0.05). There was no significant difference in the content of 5-HT,GABA,Glu,NE and MT as well as the expression of Clock and BMAL1 proteins in each group(P>0.05). Conclusion Fried Suanzaoren(Ziziphi Spinosae Semen),prepared Yuanzhi(Polygdae Padix) and Shouwuteng(Polygoni Multiflori Caulis) all have the effect of improving insomnia,among which,the high dose of fried Suanzaoren(Ziziphi Spinosae Semen) has outstanding efficacy in promoting the time of falling asleep,and prepared Yuanzhi(Polygdae Padix) has a good effect in maintaining the time of sleep;fried Suanzaoren(Ziziphi Spinosae Semen),prepared Yuanzhi(Polygdae Padix) and Shouwuteng(Polygoni Multiflori Caulis) all have significant effect of sleep aids,and the application of high dose can significantly improve the efficacy and have no obvious adverse reactions;low-dose prepared Yuanzhi(Polygdae Padix) has a clear advantage in prolonging the time of sleep. The low-dose system has obvious advantages in prolonging the duration of sleep,and at the same time,it has the effect of improving the sleep rhythm disorder,but the application of large doses can not increase the therapeutic efficacy,and has obvious adverse reactions. The tranquilizing Chinese medicines may play the role of sleep aid by promoting the synthesis and secretion of neurotransmitters 5-HT,GABA and MT,and inhibiting the CRH and Gluergic nervous system in mice.