| Objective:The study have established a senile insomnia model through D-galactose, caffeine and sleep deprivation. According to the theory of "heart is in charge of the spirit" in NeUing, through the observation of behavioral changes in rats, learning and memory, the circadian rhythm of hormone, myocardial ultrastructure, expression of peptide neurons (VIP, AVP),the study have discussed the mechanism that VIP, AVP regulated sleep-wake in rats, and Tianwang Buxin Pills (TWBXP) regulated sleep-wake's mechanisms.Methods:Male healthy SD rats were randomly allocated into four groups: normal control group,model group, TWBXP group, Estazolam group.Animals were administered with D-gal (120 mg/kg for 28 days), sleep deprivation (multi-platform water environment methods for 7days), caffeine(30 mg/kg for 7 days) to establish the senile insomnia model in rats.1. To observe the sleep, activity amount, mental state, the eyes,lips,onyx,the skin and hair.Breath frequency,heart rate,blood pressure, swimming time, the total number and the concentrstion of hemoglobin,body weight, food intake and temperature of ears was measured repectively.2. The learning and memory was measured by Morris water maze. 3. The concentration of ACTH, Cort, TSH, GH was detected through radio-immunity method, then the circadian rhythms changes were analysed by Cosinor Method.4. The myocardial ultrastructure was detected by transmission electron microscope.5.The expression of VIP,AVP both in heart and SCN was measured by immunohistochemical method and the image was analysed.6. The expression of VIPmRNA, AVPmRNA both in heart and SCN was measured by semi-quantitative analysis of a retrovirus-polymerase chain reaction (RT-PCR).Results:1. The model animals appeared insomnia symptoms, severe fatigue performance; Compared to normal control group, model group looked gray-faced and sleeping, lethargy, appetite decreased, weakness, breath frequency and heart rate quickened, blood pressure heightened,body weight and food intake decreased, temperature of ears raised, the total number and the concentration of hemoglobin decreased. TWBXP groups improved most of the symptoms, while Estazolam just improved some of the symptoms.2. In the positioning navigation experiments, compared to normal control group, model group rats' latent period of four days and two days were significantly reduced (P<0.01). In the search space experiments, the platform quadrant swimming distance percentage of the total distance was significantly falled (P<0.01), the wall 20% and 40% from the percentage of the regional swimming results showed the rising trend(P<0.01),Compared to normal control group, model group, TWBXPgroup, Estazolam group's number of crossing the location of the original platforms decreased significantly (P<0.01). 3. Compared to normal control group, model group's circadian rhythm of ACTH, Cort appeared (P<0.05,P< 0.001) while circadian rhythm of TSH,GH had not been appeared (P> 0.05, P> 0.05); Compared to normal control group,TWBXP group's circadian rhythm of ACTH,Cort,GH appeared (P <0.001, P<0.05, P<0.001) while circadian rhythm of TSH had not been appeared (P>0.05).4. Compared to normal control group, model group's myocardial ultrastructure had showed that myofilament had arranged disorderly, some of nucleus had been concentrated and diminished, chromatin had been dissolved and condensed,the structure of ridge in chromatin had been vagued, sarcoplasmic reticulum had been swelled; TWBXP group's myocardial ultrastructure had showed that myofilament had arranged disorderly, chromatin had been dissolved, the structure of ridge in chromatin had been vagued, sarcoplasmic reticulum had been swelled slightly.5. Compared to normal control group, the VTP,AVP positive cells in model group's heart had been increased significantly (P<0.01), and that in TWBXP group had been decreased significantly (P<0.01); Compared to normal control group, the VIP,AVP positive cells in model group's SCN had been decreased significantly (P<0.01), and that in TWBXP group had been increased significantly (P<0.01).6. Compared to normal control group, the expression of VIPmRNA,AVPmRNA in model group's heart had been increased significantly (P<0.01), and that in TWBXP group had been decreased significantly (P<0.01); Compared to normal control group, the expression of VIPmRNA,AVPmRNA in model group's SCN had been decreased significantly (P<0.01), and that in TWBXP group had been increased significantly (P<0.01). Conclusion:1. The multiple model therefore provides a promising model in which we can study treatments for insomnia,which has significant correlation with Yin-weakness and Blood-deficiency syndrome.2.D-gal,caffeine and SD had lead to some important changes of the cardiovascular function in animals,such as breath frequency,heart rate,blood pressure, swimming time et al,which also had resulted in the changes of myocardial ultrastructure and the expression of VIP,AVP,VTPmRNA. AVPmRNA. The multiple model had damaged "heart governed blood and vessels" function in traditional Chinese medicine theory,that had simulated the pathomechanism of "heart is in charge of spirit " disfunction in some degree, illuminated the scitific intension of "heart is in charge of spirit" which governs the sleep.3. TianWang BuXin Pills can significantly reduce the respiratory rate, depress heart rate and arterial blood pressure, extend exhaustive swimming time, increase food intake and weight gain, increase the total number and concentration of hemoglobin, decrease ear temperature, improve learning and memory, restore the circadian rhythm of ACTH, Cort and GH hormone, reduce the damage of myocardial ultrastructure,increase the expression of VIP,AVP and VIPmRNA,AVPmRNA in SCN while decrease the expression of VIP,AVP and VIPmRNA,AVPmRNA in heart.It may be illustrated by the function of TWBXP's ziyinyangxue,buxinanshen.
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