| Objective:Our aims of this study are to investigate the standardized modeling method of the premenstrual mood dysphoric disorder (PMDD) gan-qi invasion rat model, to explore the quantitative assessment of the model evaluation through the assessing the quality of the model, to probe the central mechanism under the behavior of model, studying the effect of disease and traditional Chinese medicine BXD capsule to the change of microscopic indicators in vivo.Methods:Prepared the gan-qi invasion rat model with emotional-stimulation dominated multi-factor phased stimulation (Test Ⅱ), compared with the model prepared by the emotional-stimulation dominated multi-factor continuous stimulation (Test Ⅰ), and then intervened the model with Fluoxetine or BXD capsule. To achieve the standardization of the PMDD gan-qi invasion rat model by comparing the key behavioral indicators of each group rats in Sucrose Preference Test (SPT), Open Field Test (OFT) and Elevated Plus-Maze Test (EPM).To realize the preliminary quantitative assessment of the model evaluation method ultimately, conducting the reliability test and validity estimates on the data of standardized behavioral indicators. Calculate the face validity, factor analysis parameter in construct validity, intraclass correlation coefficients (ICC) in test retest reliability, internal consistency coefficient (Cronbach’s a coefficient) of the model evalution method.After collecting the serum and separating the frontal cortex, hippocampus and hypothalamus of the rats in each group modeling under standard methods, detect the content of the monoamines (NE,5-HT, DA) in different brain regions or peripheral serum of model with high performance liquid chromatography (HPLC), and the content of the sex hormone and its regulating hormones (P, PRL, E2) content with Radioimmunoassay (RIA).Results:(1) In the review of the key behavioral indicators, on the indicator LA model group and Fluoxetine group incrsesed significantly (P<0.01、P<0.01) compared to the normal group, the BXD group decreaesd significantly (P<0.05) compared to model group, on the indicator SC%the BXD group increaesd significantly (P<0.05) compared to model group, in Test I non-receivable stage. On the indicators of OT%, OE%, SC%, model group decreaesd significantly (P<0.01、P<0.05.P<0.05) compared to the normal group, incrsesed significantly on the indicator LA (P<0.001). And they are modulated to normal apparently after intervened with Fluoxetine or BXD in Test II non-receivable stage.(2) The results of the reliability and validity of model group suggesting a good face validity for the macroscopic characterization of model group meet the clinical manifestations of the gan-qi invasion syndrome. Factor analysis showed that: evaluation of the SPT-OFT-EPM system can extract three common factors:the OT%and OE%contribute to the anxiety factor F Ⅰ (92.6%,88.6%). The LA in OFT and LA in EPM contribute to the locomotion factor FⅡ (85.2%,93.9%). The HD, Re and SC%contribute to the exploratory behavior factor FⅢ (54.1%,86.8%, and55.2%). ICC results showed that:SC%(ICC=0.959, P<0.01), LA (ICC=0.991, P<0.01), OT%(ICC=0.791, P<0.01), OE%(ICC=0.923, P<0.01) all of them have good retest reliability. The result of Cronbach’s a coefficient test suggests: Cronbach’s α=0.671in anxiety dimension, Cronbach’s a=0.254in locomotion dimension, Cronbach’s a=0.656in exploratory behavior dimension, Cronbach’s a=0.576in the evaluation system as a whole dimension. (3) Results of micro-indicators showed that: compared with the control group the content of NE in hypothalamus and frontal of model group increased significantly (P<0.05, P<0.01), decreased significantly in serum (P<0.05), the content of DA in hippocampus, hypothalamus and frontal decreased significantly (P<0.01,P<0.05,P<0.01), increased significantly in serum (P<0.01). The content of5-HT in hippocampus, hypothalamus and serum decreased significantly (P<0.05, P<0.05, P<0.05), increased significantly in frontal (P<0.05). The content of P and E2in serum decreased significantly (P<0.01, P<0.01). After intervened, compared with the model group, the content of NE in the frontal of fluoxetine group decreased significantly (P<0.05), in serum increased significantly (P<0.05), the content of DA in frontal rose significantly (P<0.05), decreased significantly in serum (P<0.05), the content of5-HT in frontal decreased significantly (P<0.01), increased significantly in serum (P<0.05).The content of P and E2in serum increased significantly (P<0.05, P<0.01). There was no significant difference in PRL content between each group. Compared with normal group, the content of NE in hypothalamus and frontal decreased significantly (P<0.05, P<0.05) in serum increased significantly (P<0.05), the content of5-HT in frontal was significantly lower (P<0.05) of the BXD group.Conclusion:(1) Compared with the emotional-stimulation dominated multi-factor continuous stimulation, the emotional-stimulation dominated multi-factor phased stimulation shows better because the principle of modeling fit more to the pathogenesis, of the preparation method ismore scientific and rational, preparation method is more scientific and rational and its behavioral indicators are closer to the group of clinical syndromes..(2) To evaluate the subtle changes in behavioral factors of the anxiety, locomotion, and exploration from the model useing SPT-OFT-EPM model evaluation method. This evaluation method has a high apparent reliability and internal consistency, showing that the evaluation methods and models fit closely. The favorable reasult of retest reliability of the model preparation methods suggests that a high degree of stability and repeatability can be owned if the gan-qi invasion rat model was prepared with emotional-stimulation dominated multi-factor phased stimulation(3) PMDD gan-qi invasion rats symptoms showing has a relationship with the concentration of NE in hypothalamus and frontal increased; the concentration of DA in hippocampus, hypothalamus and frontal decreased; the concentration of5-HT decreased in serum,hypothalamus and frontal. It is deduced that fluoxetine and BXD may exert pharmacological effects by correcting the abnormal changes in these indicators, the role of key parts is central in different brain regions. |
PDF Full Text Request