基于四氢孕酮介导大鼠杏仁核和海马脑区GABA AR亚基的经前烦躁障碍症肝气逆证发病机制研究

Study on pathogenesis of PMDD liver-qi reversal syndrome mediated by GABA ARsubunit in amygdala and hippocampus of rats based on tetrahydroprogesterone

目的观察外源性四氢孕酮(allopregnanolone,ALLO)及其抑制剂非那雄胺对经前烦躁障碍症(premenstrual dysphoric disorder,PMDD)肝气逆证模型大鼠接受期(R)和非接受期(NR)的行为学影响与GABA ARα4、GABA ARδmRNA和蛋白表达的影响,以探讨其发病机制。方法制备PMDD肝气逆证大鼠模型,将大鼠分为正常组R与NR(Control-R、Control-NR)、模型组R与NR(Model-R、Model-NR)、正常组R+ALLO与NR+ALLO(Control+A-R、Control+A-NR)、模型组R+ALLO与NR+ALLO(Model+A-R、Model+A-NR)、模型组R+非那雄胺与NR+非那雄胺(Model+F-R、Model+F-NR);运用高架十字迷宫实验和社会交互实验检测大鼠的行为学;荧光定量PCR和免疫荧光检测杏仁核和海马GABA ARα4和GABA ARδmRNA和蛋白表达。结果在行为学评价中,在NR期,在高架十字迷宫实验和在社会交互实验,模型组大鼠有焦虑行为产生以及社交能力下降(P<0.05),Model+A组能有效缓解焦虑症状和改善大鼠社会交往能力(P<0.05),Model+F组大鼠焦虑行为加重和社交障碍加重(P<0.05);荧光定量PCR和免疫荧光实验中,模型组GABA ARα4亚基在海马表达均上调(P<0.01),GABA ARδ亚基的表达均下调(P<0.01);Model+A组GABA ARα4亚基在杏仁核和海马表达降低(P<0.01),GABA ARδ亚基的表达海马脑区均升高(P<0.01)。结论ALLO可能通过介导GABA ARα4和GABA ARδ亚基的表达异常,改善PMDD的焦虑症状和社会交往能力,是PMDD肝气逆证的发病机制,为后续挖掘深层次PMDD肝气逆证的发病机制提供了依据和支撑。

Aim To observe the behavioral effects of exogenous allopregnanolone(ALLO)and its inhibitor finasteride on the receptive period(R)and non-receptive period(NR)of PMDD liver-qi inversion model rats and the expression of GABA ARα4,GABA ARδmRNA and protein effects to explore its pathogenesis.Methods The PMDD liver-qi reverse syndrome rat model was prepared.The rats were divided into the normal group R and NR(control-R,control-NR),model group R and NR(Model-R,Model-NR),normal group R+ALLO and NR+ALLO(Control+A-R,Control+A-NR),and model group R+ALLO and NR+ALLO(Model+A-R,Model+A-NR),model group R+finasteride and NR+finasteride(Model+F-R,Model+F-NR).The elevated cross labyrinth experiment and social interaction experiment were used to detect the behaviors of rats;fluorescence quantitative PCR and immunofluorescence were used to detect the expression of GABA ARα4 andδmRNA and protein in rat amygdala and hippocampus.Results In the behavioral evaluation,in the NR period,in the elevated cross maze test and in the social interaction test,the rats in the model group had anxiety behavior and decreased social communication ability(P<0.05),while the rats in the Model+A group could effectively relieve anxiety symptoms and improve their social communication ability(P<0.05),and the rats in the Model+F group had increased anxiety behavior and social disorder(P<0.05).In fluorescence quantitative PCR and immunofluorescence experiments,the expression of GABA ARα4 subunit in the model group was up-regulated in the hippocampus(P<0.01),and the expression ofδsubunit was down-regulated(P<0.01);the expression of GABA ARα4 subunit in the amygdala and hippocampus of the Model+A group decreased(P<0.01),and the expression ofδsubunit increased in the hippocampus(P<0.01).Conclusions The abnormal expression of GABA ARα4 andδsubunits mediated by ALLO improves the anxiety symptoms and social interaction ability of PMDD,which is the pathogenesis of PMDD liver-qi reverse syndrome,and provides basis and support for subsequent exploration of the pathogenesis of PMDD liver-qi reverse syndrome.