Estorgen Receptor β Mediated Serotonin System-related Genes TPH-2and Sert Nction In Rats With PMS Liver-fuql;invasion Befbre And Aiier Druu Intervention

Objective: To investigate the changes between protein location and mRNA expressionlevels of estrogen β and related genes TPH-2and SERT in brain of PMS liver-qi invasionin reception and non-reception of estrous cycles before and after drug intervertion,as wellas the possible therapeutic mechanism of traditional Chinese medicine for the disease.Methods:1. Wistar rats were chosen into experiment which with similar macroscopicbehavior indications and regular oestrous cycle examined by the vaginal smear microscopymethod and macroscopic behavior were randomly divided into four groups: control group,model group, BXD group, paeoniflorin extract group, fluoxetine group and ERβ agonistgroup. Model group rats were with conventional electrical stimulation for preparation ofPMS liver-qi invasion model.All rats were gavaged. All rats were weighed before and aftermodeling,conducted open field test scores as its macroscopic behavior evaluation.2. All rats were examined by the vaginal smear microscopy at the end of the day in order todistinguish acception and non-acception period and sample collection in correspondingperiod. The rats were separated of the brain orgnizations of hippocampus, hypothalamus,and dorsal raphe nucleus which were stored in-70℃ultra-low temperature freezerand perfused brain tissue fixation to take full after model preparation. Then we use Trizolto obtain total RNA from the brain area, and determine mRNA expression of ERβ,TPH-2and SERT with conventional RT-PCR method, and adopt the immune fluorescencedouble-tagging technology to detect the protein distribution and expression levels of ERβ, TPH-2and SERT in different estrous cycles.Results:1. Macroscopic behavior evaluation result: Before modeling, there were nosignificant differences in the initial weight in the six groups. Compared to control groupafter modeling, Model rats were significantly reduced in weight (P<0.01) whilesignificantly increased in field score (P<0.01). Meanwhile, the rats in model group looklike nervous, unstable and sensitive to surrounding stimulation. Compared to model groupafter modeling, all drug intervention groups were significantly increased in weight(P<0.05or P<0.01) and reduced in open field scores (P<0.05).2. Microscopic index measurement results:(1) In non-reception of estrous cycles,RT-PCRresults demomstrate that Compared to control group, model group was significantly lowerin ERβ and TPH-2mRNA expression (P<0.01or P<0.001), while significantly higher inSERT (P<0.001) in every cortex, Compared to model group, all drug treatment group weresignificantly higher in ERβ and TPH-2mRNA expression (P<0.01or P<0.001),whilesignificantly lower in SERT mRNA expression (P<0.01or P<0.001) in every cortex. Inreception of estrous cycles, Compared to control group, model group was no significant inERβ, TPH-2and SERT mRNA expression (P>0.05) in every cortex; Compared to modelgroup, all drug treatment group were no significant in ERβ, TPH-2and SERT mRNAexpression (P>0.05) in every cortex.(2) In non-reception of estrous cycles, Immune fluorescence tagging test demonstrate thatthe protein expression levels of ERβ and TPH-2in model group rats decreasedremarkably(P<0.01or P<0.001),while SERT increased remarkably(P<0.01or P<0.001) indorsal raphe nucleus, CA1and CA3of Hippocampus and Hypothalamus. Compared withthe model group rats, the protein expression levels of ERβ and TPH-2in all drug treatmentgroup have a significant increase(P<0.05or P<0.01), meanwhile SERT have a remarkablereduce in the every cortex(P<0.05or P<0.01). In reception of estrous cycles, Compared tocontrol group, model group was no significant in the protein expression levels of ERβ,TPH-2and SERT (P>0.05) in every cortex; Compared to model group, all drug treatmentgroup were no significant in the protein expression levels of ERβ, TPH-2and SERT(P>0.05) in every cortex. Conclusion: The bondage modeling method based on emotional stimulation is successfulin making PMS liver-qi invasion model rats. In non-reception of estrous cycles, Theabnormal changes protein levels and mRNA expressions of the ERβ, TPH-2and SERT inDorsal raphe nucleus, CA1and CA3of Hippocampus and Hypothalamus and ERK1/2protein activaty in Hippocampus may have close relationship with PMS liver-qi invasionsyndrome;and Baixiangdan Capsule can correct the low expression of ERβ,TPH-2andhigh expression of SERT in above brain regions,while the abnormal changes of relatedgenes,that probably is one of the centre pathogenesis of Baixiangdan capsule treatingPMS with liver-qi invasion syndrome.