| Objectives:On the basis of establishing the insomnia model of rats with repeated psychological stress, referring to hypnosophy and neurotransmitter index, the feasibility of psychological stress insomnia model of rats was further to be evaluated. And the effect of Jiaweixiaoyaosan on psychological stress insomnia was appraised to investigate the mechanism and provide the experimental basis for the clinical therapy.Methods: Fifty eight-week male Wistar rats, 200-250g were divided into five groups: Control Group(C), Model Group(M), Western Group(W), Herb Group(H) and Integrated Group(I). Each group (10 rats) was fixed electrode to record brain-electricity and myoelectricity. After operation a week, we made the group of M, W, H, I in restricting activity. Jiaweixiaoyaosan for H group, estazolam for W group, half of jiaweixiaoyaosan and estazolam comparing to W and H group for I group was given by lavaging respectively. Sleep-Waking cycle was monitored after restricting activity on the 1st, 3rd, 5th, 7th, 10th and 15th day respectively. After fifteen days, the expression of substance P by immunohistochemistry were detected; the ultrastructures were observed with transmission electron microscope in hippocampus and raphe nuclei. In addition, we analyze 5-HT in blood serum and raphe nuclei, NE in blood serum and hypothalamus with Highperformance liquid chromatogram.Result:1 sleep phase: 1.1 Compared with C group, the ratio of wake time increased after restriction, the ratio of non-rapid eye movement sleep and rapid eye movement sleep decreased on the 1st, 3rd, 5th, 7th, 10th and 15th day respectively. 1.2 Compared with M group, the ratio of wake time decreased respectively on the 1st, 3rd, 5th day and increased on the 7th, 10th, 15th day in W group respectively. The ratio of wake time increased on the first day and decreased on the 3rd, 5th, 7th, 10th, 15th day in H group respectively. The ratio of wake time decreased on the 1st, 3rd, 5th, 7th, 10th, 15th day in I group respectively. 1.3 Compared with M group, the ratio of non-rapid eye movement sleep increased on the 1st, 3rd, 5th, 7th day and decreased on the 10th, 15th day in W group respectively. The ratio of non-rapid eye movement sleep decreased on the first day and increased on the 3rd, 5th, 7th, 10th, 15th day in H group respectively. The ratio of non-rapid eye movement sleep increased on the 1st, 3rd, 5th, 7th, 10th, 15th day in I group respectively. 1.4 Compared with M group, the ratio of rapid eye movement sleep increased on the 1st, 5th, 7th, 10th, 15th day and decreased on the third day in W group respectively. The ratio of rapid eye movement sleep increased on the 1st, 5th, 7th, 10th day and decreased on the 3rd, 15th in H group respectively. The ratio of rapid eye movement sleep decreased on the 1st, 3rd, 5th day and increased on the 7th, 10th, 15th in I group respectively. 1.5 Numbers of wake increased after restriction,while decreased in H group. 1.6 The delta wave of non-rapid eye movement sleep decreased on the 1st, 3rd, 5th, 7th, 10th and 15th day after restriction, but increased after treatment.2 histological detectiont: 2.1 Compared with C group, hippocampus and raphe nuclei neurons'nucleus appeared chromatin condensation and margination, rough endoplasmic reticulum swelling, derangement, and ribosomal granules broken. Most of mitochondria exhibited swelling and vacuole degeneration, Golgi's complex exhibited swelling and polarity disappearance, myelin sheath broke up and synapses degenerated in most of M. The ultrastructures of H group were similarly with those of the control group in hip and RN neurons. 2.2 substance P distributed at neuron body and expressed in both nucleus and kytoplasm showing granulo-brown in C group. Compared with C group, substance P positive neurons decreased and showed in light colour in M group. The express of them reinforced in H group.3 neurotransmitter: 3.1 5-HT gently decreased in blood serum and obviously decreased in raphe nuclei in M group. After treatment with herb, 5-HT gently increased in blood serum and obviously increased in raphe nuclei.3.2 NE increased in both blood serum and hypothalamus in M group, while decreased in H group.Conclusion:1 Restriction of activity is a feasibility way of making psychological stress insomnia model of rats.2 The model of restriction of activity can cause sleep phase change in rats, evoke the ultrastructural lesion and change central neurotransmitter in raphe nuclei and hippocampus.3 Jiaweixiaoyaosan or mistura of herb and estazdam have an identified and stable effect in regulating sleep-wake cycle on the model rats. Jiaweixiaoyaosan can decrease wake time and increased NREMs. The injured raphe nuclei and hippocampus were partial recoveed and the contents of neurotransmitter were gotten close to normal by Jiaweixiaoyaosan .4 Jiaweixiaoyaosan play an important role in regulating sleep-wake cycle of the insomnia model rats by regulating the content of neurotransmitters such as 5-HT, NE and SP, and restoring the Histiooytic damage from neuroendocrine disord. |
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