| Objective:(1)The rat model of PMDD liver Qi inverse syndrome was induced by inhabitation stress,and the model of PMDD liver Qi inverse rats was identified by the method of attack behavior test: the anger attack behavior of PMDD liver Qi inverse rat model had a estrous cycle dependence;the estrous cycle dependent anger attack after the egg nest excision of PMDD liver Qi inverse rats disappeared;Exogenous sex hormone induced ovaries in PMDD rats with liver Qi reversal syndrome,the original estrous cycle dependent aggressive behavior appeared again;After drug intervention,anger attack behavior of PMDD rats with liver Qi reverse syndrome decreased or disappeared;Finally,the surface reliability,structure validity and predictive validity of the rat model of PMDD liver Qi inverse disease syndrome were identified with strict scientific evidence,and a mouse model was established to combine the disease syndrome with the mouse model.(2)Explore the underlying mechanism underlying hormone regulation through PMDD liver Qi reverse syndrome rat model,and reveal some microscopic mechanisms of liver dysfunction.It also aims to verify the new hypothesis of tutor Qiao Mingqi,that “liver catharsis is associated with monoamine neurotransmitters and sex hormones and their hormones”,to provide data support for this hypothesis,and to change the research and development of effective therapeutic drugs for PMDD liver Qi inverse syndrome in response to key biological indicators.Methods:(1)Animal behavior experiment: the rat model of PMDD liver Qi inverse syndrome was induced by habitation intrusion method,and the behavior evaluation was carried out by the methods of attack behavior test,elevated cross maze test and open field experiment.Rats underwent ovariectomy,again behavioral evaluation;The estrous cycle was induced by exogenous hormones in the model rats,and behavioral evaluation was performed again.Fluoxetine and Baixiangdan capsule intervention,behavioral evaluation again.(2)detection of neurobiochemical indexes: the key biological indexes of PMDD liver Qi inverse syndrome were detected by enzyme linked immunosorbent assay,protein immuno imprint test and polymerase chain reaction.Results:(1)The results of animal behavior test:(1)the test results of attack behavior test showed that compared with the normal group,the rats in the model group showed a large number of aggressive behavior(p<0.001)in the M,D1 and D2 stages,among which the D1 stage was the most obvious,while the E stage model group and the normal group had no difference,and there was no difference between the stage of the ovariectomized group E,M,D1 and D2(four).P>0.05);After the induction of exogenous hormone,there was a significant difference in the E phase between the model group and the M,D1 and D2 stages(P<0.01),and the most significant difference in the D1 phase.The NR phase attack in the drug intervention group decreased significantly(p<0.001),and there was no significant difference from the R phase(p>0.05).(2)the results of OFT experiment showed that compared with the normal group,the total NR period of the model rats increased significantly(p<0.001),the stay time in the central region was significantly reduced(p<0.001),and there was a significant difference(p<0.05)compared with the R stage(p<0.05).After ovariectomy,the difference between the total and central area of the NR phase disappeared(p>0.05),and there was no significant difference in NR phase and R period(p>0.05).After the induction of exogenous hormones in the model group,the difference in the total distance between the NR and the central area of the model group appeared again(p<0.001).The above changes disappeared(P>0.05),the total road course decreased significantly(p<0.001),and the stay time in the central area was significantly increased(p<0.001).There was no significant difference from the R stage.(p>0.05).(3)the results of EPM experiment showed that the rats in the model group had a significant decrease in OT% and OE%(P<0.05)during the NR phase(P<0.05),and there was a significant difference from the R phase(p<0.05);OT% and OE% increased significantly after ovariectomy and the significant difference between the reception period and the non acceptance period disappeared(P>0.05);the significant difference between the OT%,OE% acceptance and non acceptance periods was again given after the induction of exogenous hormones.In drug intervention group,OT% and OE% in NR phase increased significantly(p<0.001),and there was no significant difference compared with R stage(p>0.05).(4)the results of SIT experiment showed that in the NR phase of the model rats,the percentage of time and frequency of communication with Stranger 1 was not significantly different from that of the empty metal cage(p>0.05)in the first stage of the experiment(p>0.05)in the second stage of the exchange between the Rat2 head and the Rat2 head.In the period of R,social behavior was consistent with that in normal rats.After NR intervention,social interaction disappeared.(5)the results of SBP and HR showed that the R systolic pressure and heart rate of the model rats were basically the same as those of the normal rats,while the NR stage showed the same upward trend,but there was no significant difference(p>0.05).The systolic pressure and heart rate decreased in the NR stage after the drug intervention,which was basically close to the result of R phase.(6)the results of the width and height of the nipple showed that the width and height of the R papilla in the model rats were not different,but the width and height of the nipple were increased in the NR stage,but the width of the left nipple increased significantly(p<0.001),the right papilla width increased(p>0.05),the left papilla height and the right papilla height only showed an increasing trend(p>0.05).After drug intervention,the width and height of the nipple in NR decreased,but the width of the left nipple decreased more significantly(p<0.001).(2)The results of ELISA biochemical test showed that(1)the content of 5-HT was significantly increased in the serum,hippocampus,amygdala and frontal lobe(P<0.0001,P<0.05,P<0.01),and the content of 5-HT decreased significantly(P<0.01,P<0.05)after the intervention.(2)NE content decreased(P<0.05)and increased significantly in hippocampus,amygdala and frontal lobe(P<0.0001,P<0.01)in hippocampus,amygdala and frontal lobe,and NE content decreased significantly(P<0.001,P<0.01)after drug intervention.Although the serum NE content in Bai Xiangdan group decreased,there was no significant difference(p> 0.05).(3)the content of P in serum and three brain regions was significantly decreased(P<0.0001,P<0.001),and increased significantly after drug intervention(P<0.05,P<0.01).(4)the content of GABA decreased significantly(P<0.005,P<0.0001)in the hippocampus,amygdala and frontal lobe(P<0.005,P<0.0001),and increased significantly after the drug intervention(P<0.01,P<0.05,P<0.005,P<0.001).(5)the ALLO content in the serum and the three brain regions of the rats showed a significant decrease(P<0.01,P<0.005,P<0.0001),and the drug intervention showed an obvious trend of increase(P<0.05),and the serum of the white Xiangdan group was removed from the serum.(6)the trend of serum FSH and LH content was approximately the same.The content of NR phase decreased significantly(P<0.05),and the level of drug intervention increased significantly(P<0.05).(3)The results of PCR and Western Blot protein showed that the expression of GABAAR4 alpha protein in amygdala,hippocampus and frontal lobe was the same as that of GABAAR4 alpha m RNA,and the model group showed a significant trend(P<0.05,P<0.01),and the drug intervention group showed a tendency to decrease significantly(P<0.05).Conclusion:(1)The indignation behavior of PMDD liver Qi inverse rat model induced by inhabitation intrusion method has an emotional cycle dependence,that is,there is anger attack in the non acceptance stage of rats,with cognitive and somatoform symptoms,and the behavior of anger attack is reduced or disappeared,and the cognitive and physical symptoms disappear;After oophorectomy,the rat model of estrous cycle dependent anger attack disappeared,cognitive impairment and somatic symptoms disappeared;exogenous hormones induced the normal estrous cycle of the ovariectomized rats with PMDD liver Qi reverse syndrome,and the anger attack appeared again,that is,the estrous cycle dependence of the angry attack behavior of rats appeared again,while the normal rats were almost unaffected by the shadow of exogenous hormones.Ring;After the treatment,the anger attack behavior of the model rats weakened or disappeared,and the cognitive,social and physical symptoms improved to a certain extent.The effect of the white Dan and fluoxetine capsules was consistent,and some aspects were even better than fluoxetine.It was proved that the method of PMDD liver Qi inverse model induced by inhabitation method was stable and reliable.The model was consistent with the clinical features of PMDD liver Qi inverse syndrome,and the non acceptance period "symptoms appeared"(similar to the premenstrual period of women).The acceptance period "symptom reduction disappeared"(similar to the post menstrual follicle period)of women was in line with the primary standard of PMDD diagnosis.(2)The level of hormone changes in the model rats was basically consistent with the clinical test results,indicating that the microscopic pathogenesis of PMDD liver Qi inverse syndrome is indeed related to the hormone and monoamine neurotransmitters and their abnormal changes.The expression of 5-HT,NE,GABAAR4 alpha and m RNA is higher in PMDD liver Qi inverse syndrome,and the decrease of GABA,P,ALLO,FSH and LH is closely related.These indexes may be the key biomarkers of PMDD pathogenesis.Bai Xiangdan has a good therapeutic effect on PMDD liver Qi reverse syndrome.The effect of the treatment may be by increasing the content of P and GABA in the serum and regulating the abnormal fluctuation of 5-HT and E2 levels. |
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