| Background and Purpose:As an inhibitor of the central nervous system, ethanol can inhibit the activity of the brain cortex and cause a loss of control of the excitement of the subcortical center and brain disfunction,which then will cause a change in neurobehavioral functions, resulting in behaviorial disorders in many aspects such as learning, cognitive memory, motor coordination,etc. Therefore, excessive alcohol drinking can seriously affect one’s physical and mental health and lead to negative consequences such as traffic accidents and family violence or social violence, causing great harm to families as well as our society. But at present, many researches both at home and abroad mainly focus on the side effects of chronic alcoholism on the central nervous system, while reports on the study of acute alcohol poisoning are rather rare.This research selected adult mice as research models, which would be given different doses of alcohol intervention to establish an animal model of acute poisoning HE staining and immunofluorescence method, combined with confocal laser, will be adopted to observe the structural change of the hippocampus morphology after alcohol intervention, to discuss the acute-alchol-poisoning caused behaviorial change and its possible function mechanism in the purpose of providing a scientific basis for related researches in the future. Methods:1.Randomizing SD mice of about 250 grams, whether male or male, into control group, Low-dose group, medium-dose group and high-dose group. 2.Feeding the control group with corresponding volume physiological saline and other three groups, Red Star Wine of 56%( v/v) alcohol concentration with the ratio of 4ml/kg/8ml/kg/16ml/kg successively, and set up their respective acute alcohol poisoning models of these four groups after one hour/two hours/four hours/eight hours/twelve hours. 3. Adopingt HE staining method to observe the structural change of the hippocampus morphology after alcohol intervention.4. Adopting immunofluorescence method to observe the Neu N expressions of the hippocampal neurons at different periods after alcohol intervention.5. Using confocal laser to observe the expressions of Neu N and DAPI of Fluorescence double-stained hippocampal neurons after alcohol intervention at different periods. Findings1.The findings of HE staining show that, compared with the control group, after being fed with different ethanol concentration, no obvious changes appeared in the thickness of the cell body of the dentate gyrus neurons, while changes of different degrees appeared in the configuration of these neurons. After being fed for one hour and two hours with the ratio of 4 ml/kg, no obvioue changes appeared in the dentate gyrus neurons, slight loose appeared in the dentate gyrus granular cell configuration, to the 8th hour, granular cell configuration has been restored to normal structure. Edema appeared in the dentate gyrus granular cells of the groups fed with 8 ml/kg and ml/kg ethanol. Compared with the control group, the 8 ml/kg ethanol fed group showed no obvious difference after two hours, increaesd intersellular space and disordered and loosened cell lineage in the dentate gyrus neurons after 4 hours, returning-to-normal cell lineage structure after eight hours and equivalent density of the dentate gyrus with the control group after twelve hours. And the 16 ml/kg ethanol fed group showed visible changes in the dentate gyrus cell structure such as decreased cell density and increased intercellular space, futher injured neurons in the dentate gyrus with significantly decreased visible cell numbers and reduced tightness of the cell distribution compared with the control group. After eight hours, obviously intercellular space in the dentate gyrus disorderly scattered neurons and compeletely diaappeared layer structure are all observable and high-consentration alcohol caused serious injures in the density of The dentate gyrus neurons can be drawn. The lineage structure of the dentate gyrus granular cell did not return back to normal until twelve hours later.2.The results of the Immunofluorescence show that cone cells in CA3 and CA4 changed greatly after being fed with different doses of alcohol after hours with reduced number of cone cells and reduced hierarchy in loose configuration and increased intercellular space. After four hours of alcohol feeding, further injury to the hippocampus appeared in CA3 and CA4 and only sparse distribution of the cone cells could be observed and the layered structure configuration are totally gone. In the 16 ml/kg fed group, divergent distribution of the cells in large number could be observed in the hippocampus. After eight and twelve hours, cone cells were still loosely scattered but the numbers increased in different degrees and obvious distribution of the vacuolar cells could not be observed. Moreover, the numbers of the hippocampal neurons of the large-dose group were significantly smaller than those of thesmall-dose and medium-dose group.3.The findings of Confocal laser Neu N plus dapi show that when using nenu mark neuron nuclear, DAPI can be used to mark all kinds of nuclears.And the expressions of Neu N of the hippocampus in CA3 and CA4 began to weakn after one hour in the 4 ml/kg、8 ml/kg and16 ml/kg alcohol fed group. After two hours, the expressions of Neu N of the the dentate gyrus and hippocampus in CA3 and CA4 weakened further, the granular cell gap increased further in the structures of the dentate gyrus, he lineage of the cell got loosened and disordered, and the cone cells of the hippocampus in CA3 and CA4 decreased in numbers and disorderly scattered. The 8 ml/kgand 16 ml/kg alcohol fed group showed weaker fluorescent expression and the expression of distribution of Neun could only be observed in the dentate gyrus and part of the hippocampusin CA3 and CA4,obvious damage of the layered structure configuration of the the dentate gyrus granular cell could be onserved in spacial structure distribution, the cone cells of the hippocampus got further damaged and some cells exist in vacuolar structure to the fourth hour. And to the eight hour, the Neu N fluorescence intensity of the 4 ml/kg fed group showed no obvious difference from the control group, while the Neu N expression of the dentate gyrus of the medium-dose and large-dose group did not reture back to common and the distribution of the cells was still in loose condition. Conclusion:After ethanol lavage may result in hippocampus and dentate gyrus of the expression and distribution of Neu N changes, associated with the action time.After ethanol lavage may result in the dentate gyrus of rats and hippocampal neurons of layered structure of configuration changes, expression is loosely arranged cells, intercellular space increases, the hippocampus and dentate gyrus of pyramidal cells and granular cell configuration structure and the influence of alcohol intake is closely related to the dose and action time. |
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