| Objective: Premenstrual Dysphoric Disorder(PMDD)belongs to the category of menstrual diseases in traditional Chinese medicine.Up to now,its deep pathological mechanism still needs further study.The modern combination drugs for the treatment of this disease have large side effects and do not distinguish subtypes,so the efficacy is poor,so it is extremely urgent to find alternative treatments.Bupleurum-white peony is widely used in the clinical practice of traditional Chinese medicine.Although PMDD liver qi stagnation syndrome is an important subtype of PMDD,it is rarely reported that the two drugs treat PMDD liver qi stagnation syndrome.Therefore,based on the retrieval of liver main venting theory and experimental literature,this study intends to prepare PMDD liver qi stagnation syndrome combined with animal model and explore the role of saikosaponin and paeoniflorin(hereinafter referred to as saponin)in the treatment of this syndrome.By detecting changes in key biological indicators and hippocampal brain proteomics in model rats,we can discover the pathogenesis of PMDD liver qi stagnation syndrome and the biomarker of saponin intervention,and reveal the deep pathogenesis of liver effusion and promote the development of therapeutic drugs.Methods:(1)Through the HowNet China Journal Full-text Database,Wanfang Database and PubMed Database,literature search and theoretical research on the pharmacological effects of saikosaponin and paeoniflorin,premenstrual dysfunction disorders and liver stagnation;(2)first use forcing The Swimming Swimming Test(FST)and Tail Suspension Test(TST)evaluated the effect of saikosaponin in combination with paeoniflorin on mice treated with depression-like mood.(3)Combined with the pregnancy analysis method,the rat model of PMDD liver qi stagnation was prepared by forced swimming stress screening,and the model rats were performed by FST,Elevated Plus-Maze(EPM)and Splash Test.Behavioral evaluation.(4)Rats who met the standard of rat model of PMDD liver qi stagnation syndrome were treated with three doses of saikosaponin and fluoxetine.After the intervention,the above behavioral rats were evaluated again to verify the treatment of PMDD liver qi stagnation.The effect of the card.(5)ELISA(Enzyme-linked immunosorbent assay,ELISA)was used to detect P,ALLO,E2 and other biological indicators in the peripheral blood,hypothalamus and hippocampus of the model rats,and the content changes between the groups were compared.(6)TMT relative quantitative proteomics technique was used to study the hippocampal brain area of rats with PMDD liver qi stagnation syndrome.The samples were detected by Q ExactiveTM HF-X mass spectrometer in data dependent acquisition mode.The tandem mass spectrum was analyzed by PEAKS Studio version X(Bioinformatics Solutions Inc,Waterloo,Canada).The protein expression of hippocampus in rats with PMDD liver qi stagnation syndrome was analyzed by GO analysis and KEGG analysis.Results:(1)Pharmacodynamic study of mice: Through FST and TST experiments in mice,it was found that there were obvious changes in each administration group compared with the blank control group.Compared with the blank control group,the suspension immobilization time of fluoxetine group decreased in three experiments,and the second experiment showed that the suspension immobilization time of fluoxetine group decreased significantly(P<0.05,P<0.01).Compared with the blank control group,the suspension immobilization time of the high-dose group also showed the same decreasing trend in three experiments,and the second experiment and the third experiment showed that the suspension immobilization time of the high-dose group was significantly lower than that of the blank control group(P<0.05,P<0.01).The low-dose group showed a decreasing trend in the suspension immobilization time of the blank control group,and the third experiment showed that the suspension immobilization time of the low-dose group There was significant difference between the two groups in the second experiment(P < 0.01).(2)Through forced swimming experiment and regular detection of estrous cycle,a rat model with PMDD Liver-qi Depression Syndrome was successfully prepared,which was characterized by regular estrous cycle and "symptoms appeared before menstruation and disappeared after menstruation".There was a significant difference in suspension immobility time between non-acceptance period and acceptance period(P < 0.05,P < 0.05).01).The results of the elevated cross maze test and the sprinkler test both confirm the validity of the PMDD Liver-qi Depression Model prepared by FST.After treatment with saikosaponin and fluoxetine,the suspension immobilization time,suspension immobilization times and modification time in sprinkler experiment of the model group rats changed significantly.(3)The results of ELISA suggested that the depressive state of PMDD rats with Liver-qi Depression was highly correlated with the changes of ALLO,P and E2 contents in peripheral serum and hippocampal brain.The decrease of ALLO content was significantly affected by the decrease of P.The decrease of E2 content in serum of model rats may be one of the pathogenesis of PMDD with liver-qi depression.Saikosaponin can improve the symptoms of PMDD with Liver-qi Depression by increasing the levels of ALLO,P,E2 in serum,ALLO in hippocampus and P in hypothalamus.(4)Proteomic results showed that compared with the model group,70 differential proteins were found in the normal control group,260 differential proteins were found in the low dose group of saikosaponin,and 189 differential proteins were found in the fluoxetine group.Compared with the normal control group,277 differential proteins were found in the saikosaponin group and 228 differential proteins were found in the fluoxetine group.Compared with the fluoxetine group,only four different proteins were found in the saikosaponin group.Through the analysis of KEGG PATHWAY results,we can find that the main pathways involved in PMDD rat model of Liver-qi Depression are serotonin synaptic pathway,dopaminergic synaptic pathway,progesterone-mediated oocyte maturation pathway and neurogenesis signaling pathway.Conclusion:(1)Chinese medicine believes that the disease is mainly in the liver and the disease is complicated and complicated.Therefore,the treatment is mostly from the liver and the cause is due to liver loss.The use of Bupleurum-white peony is not effective in treating liver qi stagnation.The classic prescription of a syndrome is widely used in clinical practice.Therefore,it is of great research value to use the two components of Chaiqi to treat PMDD liver qi stagnation syndrome.The behavioral test of mice in this study showed that the depression-like mood of the dried saponins of the saponins of saponins had a significant improvement effect,while the rat experiment showed that the saponins of saponins had a good therapeutic effect on the rat model of PMDD liver qi stagnation.(2)The pathogenesis of the rat model of PMDD liver qi stagnation is closely related to the changes of progesterone,tetrahydroprogesterone and estradiol in peripheral blood and some brain regions.The saponin can play a therapeutic role by up-regulating the above-mentioned changes in hormone content.(3)Proteomics results suggest that the pathogenesis of PMDD liver qi stagnation may be related to the expression of Prkaca,Kcnma1 and Ywhah.The down-regulation of Kcnma1 protein suggests that the regulation of calcium channel activation,the up-regulation of Prkaca protein suggests the regulation of cAMP signaling pathway,and the upregulation of Ywhah protein.This suggests that phosphoserine binding protein mediates signaling.However,the saponin can reverse the expression of the above proteins through multi-pathway and multi-target therapeutic effects.Therefore,we speculate that the pathogenesis of PMDD liver qi stagnation syndrome may be related to the expression of Ywhah and Prkaca proteins in the hippocampus,and the above proteins are used as The diagnostic biomarkers of PMDD liver qi stagnation syndrome and the therapeutic targets of saponin saponins deserve further in-depth verification and exploration. |
PDF Full Text Request