Study On The Underlying Mechanism Of Disorders Caused By The Liver Failing To Convey And Disperse-Pathogenesis Of PMDD Liver-Qi Depression From The Change Of ALLO And GABA_AR4? Subunit Gene And Protein Expression

Objective: Through reviewing the theory and experiental literature of the liver conveying and disperseing systematically,we prepare the premenstrual dysphoric disorder(PMDD)rat model of liver qi depression in rats by Forced Swim Stress.And to detect the key biomarkers of the peripheral blood and brain of the model rats in order to screen and lock the biomarkers closely related to the pathogenesis of PMDD liver qi depression,and to provide valuable clues for the development of targeted therapeutic drugs and first-hand empirical evidence to support the possible central mechanism of liver failing to convey and disperse.Methods: This study used animal behavioral experiments and neurobiological indicators of the combination of methods to explore.?The rats in the model of PMDD liver qi depression were induced by forced stress(3times :D1,R,D1),the immobility time,the elevated plus maze,the Y maze,the feeding rate test are used to evaluate the model,in order to provide effective model of PMDD liver qi depression;? After the intervention of Fluoxetine and ShuYu capsule,the behavior above are improved,one more time to verify the reliability of the animal model;? by ELISA kits in serum and part of the brain(Amygdaloid nucleus,Hippocampus,Prefrontal)5-HT,NE,GABA,E2,P,ALLO index changes in each group,using rt-pcr,western blot method to detect GABAAR4 alpha gene and protein expression,to observe the Change of ALLO and GABA_AR4? subunit gene and protein expression.Results: (1)The results of animal behavior test :FST showed that the immobility time and the immobility number were significantly increased(P<0.05,P2<0.05),and EPM test showed that the percentage of OT%(percentage of open arm entering the elevated cross maze)in the stress+water group was significantly lower than that of the control group(P>0.05),After the intervention of Fluoxetine and ShuYu capsule,the change was disappeared.The above behavioral parameters of the mice have a typical estrous cycle-dependent change,which is similar to or consistent with the clinical characteristics of the disease,once again confirmed that the forced swimming-induced PMDD liver qi dispression rat model has good surface validity and expected validity.(2)The results of biochemical tests : the changes of 5-HT,NE and ALLO in serum,hippocampus,amygdala and frontal lobe of rats were relatively the same,and the stress+water group was significantly decreased(P<0.05,P<0.01),the administration group was significantly increased(P<0.05,P <0.01);P in serum and three brain regions,stress + pure water group was significantly increased(P<0.05),the treatment group was significant(P<0.05,P<0.01).Except for Shuyu administration group,the content of progesterone was lower than that of stress+water group,but no statistical difference was found.The E2 content in stress+water group(P<0.01).There was a decreasing trend in the two groups after drug intervention.(P<0.05),and there was significant difference between the two groups(P<0.05,P<0.01),and the levels of GABA in the three groups were significantly decreased(P<0.05)),Stress + fluoxetine administration group increased significantly more stress +Shuyu administration group,which shows that the incidence of PMDD liver qi stagnation and abnormal changes in the above indicators are closely related;(3)RT-PCR and Western Blot protein: the expression of GABA_AR4? protein and mRNA in amygdala,hippocampus and frontal lobe of rats were similar,and the stress+water group was significantly higher than that of blank control group(P<0.01,P<0.05),suggesting that abnormal expression of GABA_AR4? subunit may be the effect of PMDD liver qi(P<0.05),stress+fluoxetine group and stress+Shuyu group compared with stress+pure water group,it may be one of the pathogenesis of PMDD Liver-Qi DepressionConclusion:(1)The model of PMDD Liver-Qi depression was more consistent with the primary criteria for the diagnosis of PMDD,and the immobility time and the OT% has significant estrous cycle dependency which increase the cognitive and somatic symptoms of the model rats suggesting a good surface validity,structural validity,expected validity;(2)The expression of E2,P,GABA_AR4? protein and mRNA are high,and the contents of NE,5-HT,ALLO and GABA are low,which were closely related to the pathogenesis of PMDD.So these markers may be the key biomarkers of Liver-Qi depression.(3)The reason why the Liver failing to convey and disperse is highly correlated with the above abnormalities.The Change of ALLO and GABA_AR4? subunit gene and protein expression may be the key to the deep mechanism.ALLO can bind directly to GABAAR in the brain.First,through the opening of the Cl-channel on GABA,Which inhibits the excitability and inhibition of neurons by inhibiting the inhibition of GABA on the HPA axis and the regulation of GABA uptake,resulting in a variety of hepatic evading disorders.