Study On Shuxin Anshen Paste’s Clinical Observation And Mechanism On Insomnia For Yang-deficiency Constitution

BackgroundInsomnia is a common clinical manifestation for many physical and mental illness, which normal sleep/wake rhythm is disrupted. Persisting insomnia can make social function and quality decline. With the development of lifestyle changes and life stress, insomnia crowd is gradually increasing.45.4%of people experienced different degrees of insomnia in the past month in China, according to global epidemiological investigations in2002,30%-50%of adults have symptoms of insomnia in France in1980-2009, in2011, the American insomnia survey showed that the prevalence of insomnia was22.1%. The constitution is closely related to insomnia, biased individuals is characteristic of qi, blood, Yin and Yang are imbalanced, which cause rhythm of sleep disorders. Insomnia is accounted for16.61%of the survey of8438cases of medical and health of the population Key Projects in the National Science&Technology Pillar Program during the Eleventh Five-Year Plan Period, the most common constitution of is Yang deficiency. Insomnia treatment has the vital significance judge from Chinese medicine constitution.At present, most of drugs in treatment of insomnia is sedative hypnotics, anti-anxiety, anti-depression, and hormone therapy. However, these drugs has some adverse effects in the process of taking medicine, Such as degrading memory, fatigue weakness, dependence and withdrawal reaction, etc. Modulation of insomnia has a unique advantage for Chinese medicine which can regulate the body’s balance by the way of the natural properties of Chinese herb. Paste is the main way to improve Chinese medicine constitution, its purpose is to balance the qi and blood, Yin and Yang to restore the rhythm of sleep, conforming to the trend of the medical model of biological-society-mental development. Objectives1. To assess the clinical effect of Shuxin Anshen Paste’s in treatment of insomnia by clinical features and assessment scale, to figure out effects on the human constitution and survival quality, and then, the paste is widely used in patients.2. To study acute toxicity, the sedative, hypnotic effects and the effect on sports coordination of Shuxin Anshen Paste so that evaluate the safety and pharmacodynamics of Shuxin Anshen Paste.3. To study effects of Shuxin Anshen Paste on the monoamine neurotransmitters in rat brain based on the sleep deprivation model High-performance with liquid chromatography with electrochemical detection(HPLC-ECD) to study on sedative hypnotic effect of the paste.4. To study the effect of Shuxin Anshen Paste on zebrafish behavior with video record and track system in order to study Chinese herb drug excited/inhibition effect.Methods1. Clinical effects observation:We evaluated the clinical curative effect of using Chinese medicine health scale, Pittsburgh sleep index scale, severe insomnia index, anxiety self-assessment scale, depression self-rating scale and the world health organization quality of life rating scale from the internal medicine outpatient Guangdong Provincial Hospital of Chinese medicine. We evaluate on symptoms change, health status, depression, anxiety and the quality of life before the intervention, the first month, the second month and the third month, input ing into Epidata3.0database and andanalyzing of the data by the way of spssl8.0.2. Acute toxicity and pharmacodynamics study:We continuously observe the adverse reaction and death of NIH mice with maximum concentration and maximum volumetric dosing within14days, recording the animal death, weight, skin color, respiratory system and nervous system symptoms. Effect on the motor activity to the open field and synergetic effect with pentobarbital sodium were observed, the effect of Shuxin Anshen paste on the muscle coordination was determined by rota-rod system.3. Study on eurotransmitter mechanism:Insomnia model was established on rats by means of parachlorophenylalanine (PCPA) abdominal injection. High-performance liquid chromatography with electrochemical detection(HPLC-ECD) was used to determined5-hydroxytryptamine (5-HT),5-hydroxyindole acetic acid(5-HIAA), dopamine(DA) and norepinephrine(NE) in the brain stem and cerebral cortex.4. Study on modeling behavior in Zebrafish:Zebrafish were exposed to different concentrations of paste and Chinese herb, their behavioral indicators in the tank, including track diagram, movement speed, length, time were recorded and quantified automatically with the video record and track system, watching the movement of zebrafish activity under the photoperiod stimulus.Results1.The total effective rate was respectively32.5%,42.5%,63.5%in the first month, in the second month and in the third month after taking medicine. Compared to before taking-medicine, Insomnia seriously index, Anxiety self-assessment scores, and Depression self-rating scores decreased significantly (P<0.01), Physiological field, Psychological field scores increased significantly (P<0.05), Social relations, Environment were not significantly different respectively (P>0.05)2.Shuxin Anshen Paste could enhance the sleeping number of pentobarbital sodium and prolong the sleeping time of sodium pentobarbital, it also decreased the falling rate of mice from the roat-rod test in contrast with the positive reference drugs(diazepam).3. Precise separation of the four neurotransmitters was achieved using isocratic elution in9minutes, a good liner response was obtained over the range of1-1000ng· mL-1with correlation coefficients not less than0.997. The absolute recoveries was not less than95.56%;Shuxin Anshen Paste could significantly decrease the contents of5-HT,5-HIAA and NE inside rat’brain stem and cerebral cortex,but it had no influence on the contents of DA.4. Locomotor activity assay showed the average speed of larvae under dark condition was significantly higher than that of light condition. As the paste concentration increases, the larval locomotor activity gradually reduced. Under the condition of illumination, low dose group can enhance larval locomotor activity (P<0.05), medium and high dose group can reduce larval locomotor activity, but there is no statistical significance. Under the condition of dark, medium and high dose group can reduce larval locomotor activity (P<0.05) Under the condition of illumination, the locomotor activity of ramulus cinnamomi decoction, fossilia oyster decoction and fructus cornel decoction are significantly higher than control group (P<0.05); Under the condition of dark, the locomotor activity of sini decoction are significantly higher than control group (P<0.05), the locomotor activity of sini guigan longmu decoction and ramulus cinnamomi decoction significantly lower than the control group (P<0.05)Under the condition of illumination, aconite root, amomi, fructus and cornel can significantly enhance the locomotor activity of zebrafish(P<0.05). Under the condition of dark, zingiberis amomi can significantly reduce the locomotor activity (P<0.05),while aconite root, fructus and cornel can significantly enhance the locomotor activity (P<0.05)Conclusion1. The curative effect of Shuxin Anshen Paste is gradually enhanced perhaps because of drug cumulative effect. It can not only improve sleep (sleep quality, Sleep time, sleep time, sleep efficiency, etc.), and also can improve the subjective sleep quality, physiological and psychological.2. Shuxin Anshen Paste shows hypnotic and sedative effects in rats, but does not impair sports coordination.3. It was a more reliable and sensitive method to mesure the neurotransmitters in rat brain using HPLC-ECD;The NE level of brain may be play the main role in treating insomnia by adjusting the level of5-HT,5-HIAA and NE to restore normal sleep rhythm.4. Shuxin Anshen Paste can obviously reduce larval movement speed, and do not change the cycle of zebrafish regularity. As the paste concentration increases, the larval locomotor activity gradually reduced. Under the condition of illumination, ramulus cinnamomi decoction, fossilia oyster decoction, fructus cornel decoction, aconite root, amomi, fructus and cornel can significantly enhance the locomotor activity of zebrafish. Under the condition of dark, sini decoction, aconite root, fructus and cornel can significantly enhance the locomotor activity; sini guigan longmu decoction, ramulus cinnamomi decoction, zingiberis amomi can significantly reduce the locomotor activity.