【摘要】 目的：将白香丹胶囊作用于经前烦躁障碍症（PMDD）肝气逆证模型大鼠，从四氢孕酮（ALLO）介导海马脑区γ-氨基丁酸A型受体（GABAAR）α4、GABAARδ亚基的表达变化，阐述白香丹胶囊对于PMDD肝气逆证的作用机制。方法：运用激素诱导大鼠动情周期联合居住入侵诱导PMDD肝气逆证模型大鼠，将大鼠分为对照组、模型组以及白香丹低、中、高剂量组，每组各10只，灌胃给药12 d(3个动情周期)，采用居住入侵、高架十字迷宫、社会交互实验观察各组大鼠行为学特征；采用酶联免疫吸附测定（ELISA）测量大鼠血清中ALLO的含量；采用免疫荧光观察大鼠海马脑区GABAARα4、GABAARδ蛋白表达水平；采用实时荧光定量聚合酶链反应（qRT-PCR）检测海马脑区GABAARα4、GABAARδmRNA表达水平，综合分析白香丹胶囊对于PMDD肝气逆证的作用机制。结果：高架十字迷宫实验结果显示，与对照组比较，模型组大鼠进入开放臂时间的百分数（OT%）、进入开放臂次数的百分数（OE%）显著降低（P<0.05或P<0.01），与模型组比较，白香丹低、中、高剂量组大鼠OT%均显著升高（P<0.05或P<0.01），白香丹高剂量组OE%显著升高（P<0.05）。社会交互实验结果显示，与模型组比较，白香丹高剂量组进入陌生鼠箱时间显著延长（P<0.05）；居住入侵实验结果显示，白香丹中剂量组打斗得分显著低于模型组（P<0.05）。ELISA检测结果显示，与对照组比较，模型组ALLO含量显著下降（P<0.001）；与模型组比较，白香丹低、中、高剂量组ALLO含量显著增加（P<0.001）。免疫荧光检测结果显示，与对照组比较，模型组大鼠海马脑区GABAARα4蛋白表达显著增加，GABAARδ蛋白表达显著降低；与模型组比较，白香丹低、中、高剂量组GABAARα4的蛋白表达均显著降低，而GABAARδ的蛋白表达均显著升高。qRT-PCR结果显示，与对照组比较，模型组大鼠海马脑区GABAARα4mRNA表达水平显著升高（P<0.05）,GABAARδmRNA表达显著降低（P<0.001）；与模型组比较，白香丹低剂量组大鼠海马脑区GABAARα4mRNA表达水平显著下降（P<0.01），白香丹高剂量组GABAARδmRNA表达显著升高（P<0.001）。结论：白香丹胶囊能够提高PMDD模型大鼠血清中ALLO含量，调控模型大鼠海马脑区中的GABAARα4、GABAARδ亚基表达，有效改善PMDD模型大鼠非接受期烦躁易怒的典型行为学表现。ALLO介导的海马脑区GABAARα4、GABAARδ亚基表达变化是白香丹胶囊干预PMDD肝气逆证的重要作用机制。更多还原

【Abstract】 Objective:To study the effect of Baixiangdan Capsule（白香丹胶囊） on rats with the counterflow of liver qi syndrome of premenstrual dysphoric disorder(PMDD),and to elucidate the mechanism of action of Baixiangdan Capsule on the counterflow of liver qi syndrome of PMDD from the expression changes of allopregnanolone(ALLO) mediated γ-aminobutyric acid type A receptor(GABAAR) α4 and GABAARδsubunits in Hippocampus. Methods:The rat models of the counterflow of liver qi syndrome of PMDD were established by Hormone induced estrous cycle and co-habitation invasion. The rats were divided into control group,model group,and low-dose,medium-dose and high-dose groups of Baixiangdan,10 for each group.Baixiangdan were administrated by intragastric administration for 12 days(3 estrous cycles). The behavioral characteristics of the rats were observed by residential invasion,elevated cross maze and social interaction tests. The content of ALLO in serum was measured by enzyme-linked immunosorbent assay(ELISA). The protein expression levels of GABAAR α and GABAARδ in the hippocampus of rats were observed by immunofluorescence. The mRNA expression levels of GABAARα and GABAARδ in hippocampus were detected by realtime fluorescence polymerase chain reaction(qRT-PCR),and the mechanism of action of Baixiangdan Capsule on the counterflow of liver qi syndrome of PMDD was comprehensively analyzed.Results :In the behavioral detection,the results of the elevated cross maze experiment showed that compared with the control group,the percentage of enter open arm time(OT%) and percentage of enter open arm frequency(OE%) of entering the open arm of rats in the model group were significantly decreased(P<0.05 or P<0.01). Compared with the model group,OT% of rats in the low,medium and high dose groups of Baixiangdan was significantly increased(P<0.05 or P<0.01),and OE% of rats in the high dose group of Baixiangdan was significantly increased(P<0.05). The results of the social interaction experiment showed that the time spent in the unfamiliar rat box of the high dose group of Baixiangdan was significantly longer than that of the model group(P<0.05). The residential invasion experiment showed that the fighting scores of the medium dose group of Baixiangdan were significantly lower than those of the model group(P<0.05).The results of the ELISA detection showed that compared with the control group,the content of ALLO of the model group was significantly lower(P<0.001). Compared with the model group,the content of ALLO of the low,medium,and high dose groups of Baixiangdan was significantly increased(P<0.001). The results of the immunofluorescence detection showed that compared with the control group,the protein expression of GABAARα4 in the hippocampus of the model group was significantly increased,while the protein expression of GABAARδ was significantly decreased. Compared with the model group,the protein expression of GABAAR α 4 in the low,medium,and high dose groups of Baixiangdan was significantly decreased,while the protein expression of GABAARδ was significantly increased. The qRT-PCR results that compared with the control group,the mRNA expression level of GABAARα4 in the hippocampus of the model group was significantly increased(P<0.05),and the mRNA expression of GABAARδ was significantly decreased(P<0.001). Compared with model group,the mRNA expression level of GABAARα4 in the hippocampus of the low dose group of Baixiangdan was significantly decreased(P<0.01),and the mRNA expression level of GABAAlδ in the high dose group of Baixiangdan was significantly increased(P<0.001). Conclusions:Baixiangdan Capsule may improve the typical behavioral manifestations of non-receptive period of PMDD model rats by increasing the content of ALLO in serum and regulating the expression of GABAARα4 and GABAARδ subunits in the hippocampus of model rats. Therefore,the expression changes of GABAARα4and GABAARδ subunits in the hippocampus mediated by ALLO may be the mechanism of action of Baixiangdan Capsule in the intervention of the counterflow of liver qi syndrome of PMDD.更多还原