Analysis Of TCM Syndrome Characteristics Of Postpartum Depression And Research On The Intervention Mechanism Of Xiaoyao Powder

Objective:1.Clinical epidemiological studyTo explore the influencing factors of aggravation of postpartum depression(PPD)by clinical cross-sectional epidemiological investigation.In addition,the factors and cluster analysis were used to explore the traditional Chinese medicine(TCM)syndrome characteristics of PPD,and further summarized the pathogenesis and treatment of PPD,in order to provide theoretical support for the clinical individualized accurate differentiation of PPD.2.Research on TCM literature and prescriptionTo explore the drug composition rules and medication ideas of the treatment of PPD by searching the relevant literature of clinical research on the treatment of PPD with traditional Chinese medicine,in order to provide a reference for the clinical treatment of PPD and a theoretical basis for the further research on the mechanism of TCM in the treatment of PPD.3.Animal experimental mechanism researchCombined with the clinical epidemiology of PPD and the research of TCM literature and prescription,animal experiment was carried out to establish and evaluate the PPD rat model,and Xiaoyaosan was given after the success of the PPD model.To explore the therapeutic effect and molecular mechanism of Xiaoyaosan in the treatment of PPD by observing the pathological morphology of hypothalamus,detecting the hormone level of HPG axis and the expression level of PKA/CREB/BDNF signal pathway mediated by hypothalamic estrogen receptor(ERa,ER?,GPER)of rats,in order to provide objective basis with scientific connotation for Xiaoyaosan in the treatment of PPD,and at the same time to reveal the TCM syndrome characteristics and pathogenesis essence of PPD based on 'syndrome detecting from prescription used'.Methods:1.Clinical epidemiological studyThe Questionnaire of PPD was established to collect the general situation,depression scale score,TCM symptoms and signs information of PPD patients.The Pearson chi square test or Fisher exact probability method were used to analyze the general situation of patients.The logistic regression of ordered multi classification was used to analyze the statistical related factors in the results of single factor analysis and to explore the factors that led to the aggravation of depression in PPD patients.The clinical symptoms and signs of patients with PPD were studied by frequency analysis,factor analysis and cluster analysis.The TCM Syndromes of PPD patients were summarized,and further explore the pathogenesis and treatment principles of PPD.2.Research on TCM literature and prescriptionSearch the relevant literature on PPD clinical research of TCM published in 2000-2019 by CNKI,Wanfang database and other domestic Chinese databases.The Chinese medicine prescription needed for the study was collected according to the screening criteria.The SPSS 24.0 and Weka 3.8 were used to analyze the frequency analysis,cluster analysis and association rules analysis of TCM after the establishment of database,so as to explore the rule of prescription and medication of TCM in the treatment of PPD,and explore the medication ideas of clinical treatment of PPD.3.Animal experimental mechanismPPD rats were established by Chronic ultramild stress(CUMS)method in the late pregnancy period.The model was evaluated by the general situation,weight,litter size,live litter size and behavioral changes of rats.After the model was successfully established,HE staining was used to observe the pathological morphology of hypothalamus of rats,and serum E2,P and PRL were detected by ELISA.RT-qPCR and Western blot were respectively used to detect the gene and protein expression levels of ERa,ER?,GPER,PKA,CREB and BDNF in the hypothalamic tissue homogenate.The expression of molecular protein was analyzed by immunohistochemistry method.The pathogenesis of PPD and the intervention mechanism of Xiaoyaosan were explored based on HPG axis.At the same time,the biological basis of PPD syndrome of stagnation of liver qi and spleen deficiency was revealed by 'syndrome detecting from prescription used' and Xiaoyaosan as the evidence.Results:1.Clinical epidemiological study(1)The average age of 206 patients with PPD was 30.28±5.07 years old;94 patients with mild depression,accounting for 45.63%;72 patients with moderate depression,accounting for 34.95%;40 patients with severe depression,accounting for 19.42%.Among the psychosocial factors influencing the aggravation of depression in PPD patients,the prenatal education was the protective factor.Neonatal disease,premenstrual irritability,anxiety and depression during pregnancy,negative events during pregnancy and childbirth,lack of sleep time,low degree of care and support of patients were the risk factors for the aggravation of depression in patients with PPD.(2)In the investigation of TCM symptoms and signs of 206 patients with PPD,the high-frequency symptoms and signs were emotional depression,insomnia and dreaminess,fatigue,anorexia,irritability,sighing,distending pain of breasts,sadness and crying,energielos,forgetfulness,abdominal distension,light red tongue,teeth-printed tongue,dark tongue,ecchymosis or petechia on tongue,white fur,thin fur,greasy fur,thready pulse,string pulse and deep pulse.The 9 common factors of TCM symptom factor analysis were analyzed by cluster analysis.There are 5 clustering results.? syndrome of stagnation of liver qi and spleen deficiency,its symptoms include sighing,abdominal distension,fullness of flank ribs,hypochondriac distension or even pain,distending pain of breasts,anorexia;? syndrome of yin blood deficiency,its symptoms include pale complexion,light nail color,numbness of limbs,tidal fever and night sweating,dysphoria with feverish sensation in chest,palms and soles,dizziness and tinnitus;?syndrome of exuberance of liver fire,its symptoms include bitter taste in mouth,red eyes,dry mouth and throat,constipation,irritability and headache;?syndrome of yang deficiency of spleen and kidney with phlegm dampness,its symptoms include epigastric oppression,nausea,oppression in chest,panic awaken,fear of cold and chilly limbs,soreness and weakness of waist and knees,edema,clear urine in large amounts or(and)increased nocturia;?syndrome of deficiency of both heart and spleen with blood stasis,its symptoms include palpitation,fatigue,shortness of breath and languid speech,energielos,pale complexion,forgetfulness,insomnia and dreaminess,sad and crying,dark complexion,spontaneous sweating,lower abdominal tingling,dark purple lochia with mass.2.Research on TCM literature and prescription(1)112 prescriptions were selected in this study,involving 116 kinds of Chinese herbs,and the frequency of use was 1216.High frequency drugs for PPD include Bupleuri Radix,Glycyrrhizae Radix et Rhizoma,Angelicae Sinensis Radix,Paeoniae Radix Alba,Poria,Atractylodis Macrocephalae Rhizoma,etc.Among all the drugs,spleen meridian and liver meridian are the main channel tropism,and warm and sweet drugs are the most frequently used.The drug efficacy is mainly supplementing,regulating qi,tranquilizing,promoting blood circulation for removing blood stasis.(2)17 high-frequency drugs were divided into four groups:? cluster 1 included Bupleuri Radix,Paeoniae Radix Alba,Angelicae Sinensis Radix and Glycyrrhizae Radix et Rhizoma;? cluster 2 included Atractylodis Macrocephalae Rhizoma and Poria;? cluster 3 included Cyperi Rhizoma,Citri Reticulatae Pericarpium,Chuanxiong Rhizoma and Curcumae Radix;? cluster 4 included Poria,Astragali Radix,Jujubae Fructus,Rehmanniae Radix,Albiziae Cortex,Ziziphi Spinosae Semen,Polygalae Radix.(3)A total of 50 association drug combinations were obtained from the association analysis of 17 high-frequency drugs.6 groups met the second-order association rules,18 groups met the third-order association rules,and 17 groups met the fourth-order association rules.3.Animal experimental mechanism(1)Experiment 1:there was no significant difference in litter size and live litter size among the four groups.After modeling,rats showed signs of fatigue and drowsiness,decreased activity,soft feces,weight loss,and decreased sugar consumption rate and forced swimming immobility time in behavioral results.In open field test,the number of squares crossed,rearing and grooming showed a decreasing trend.In Fluoxetine group and Xiaoyaosan group,the macroscopical and behavioral results of rats were significantly improved,which indicated that the postpartum depression like changes occurred in the model group.The PPD rat model was successfully established.The PPD rat model established in this study is a combination model of PPD with syndrome of stagnation of liver qi and spleen deficiency by 'testing syndrome by prescription' and Xiaoyaosan as the evidence.(2)Experiment 2:in the HPG axis,HE staining showed that the number of hypothalamic neurons in the model group decreased,the arrangement was loose and disordered,a large number of neurons appeared nuclear pyknosis and deep staining,cell body atrophy,vacuolar degeneration and peripheral demyelination changes and other serious pathological changes But the histopathological changes of hypothalamus in rats of Xiaoyaosan group were improved in varying degrees.ELISA detection showed that the serum E2,P,PRL levels of the model group were significantly decreased.Compared with the model group,the serum E2,P,PRL levels of the Xiaoyaosan group were significantly increased.(3)Experiment 3:RT-qPCR results showed that the gene expression levels of ERa,GPER and PKA in hypothalamus were significantly decreased in model group than in normal group,while the gene expression levels of ER?,CREB and BDNF had decreased trend.Compared with the model group,the gene expression levels of ERa,CREB and BDNF in hypothalamus of rats in Xiaoyaosan group were significantly increased,the gene expression levels of ER?,GPER and PKA had increased trend.Western blot results showed that the protein expression levels of ER?,CREB and BDNF in hypothalamus were significantly decreased in model group than in normal group,and the protein expression levels of ER?,GPER and PKA had decreased trend.Compared with model group,the protein expression levels of PKA and BDNF in hypothalamus of Xiaoyaosan group were significantly increased,and the protein expression levels of ERa,ER?,GPER and CREB were increased.Immunohistochemical localization analysis of the above indicators showed that the immunopositive neurons were widely distributed in hypothalamic arcuate hypothalamic nucleus(ARC)and ventromedial hypothalamic nucleus(VMH).The results showed that the MOD value of ER?,PKA,CREB and BDNF in hypothalamic ARC of model group were significantly decreased,while the MOD value of ER?,ER?,GPER,CREB and BDNF in VMH were significantly decreased.Xiaoyaosan could reverse this change and significantly increase the expression level of the related indexes.Conclusion:1.Psychological and social factors interact with each other,which can be used as stressors to lead to the occurrence and development of PPD.Syndrome of stagnation of liver qi and spleen deficiency,syndrome of yin blood deficiency,syndrome of exuberance of liver fire,syndrome of yang deficiency of spleen and kidney with phlegm dampness,syndrome of deficiency of both heart and spleen with blood stasis and their clinical symptoms are the TCM syndrome characteristics of PPD patients with clinical significance2.Women postpartum deficiency and blood stasis,imbalance of qi and blood and malnutrition of spirit and soul exist in the onset of PPD.Based on the close correlation between liver and spleen,emotion,qi and blood,clinical treatment of PPD patients can be treated from liver and spleen,regulate qi and blood,clinical flexible use methods of dispersing stagnated liver qi for relieving qi stagnation,regulating qi-flowing for strengthening spleen,replenishing qi and nourishing blood,nourishing heart for tranquillization and other therapies.Xiaoyaosan is a common prescription for the treatment of PPD,which combines qi and blood and harmonizes liver and spleen.3.The rat model of PPD with syndrome of stagnation of liver qi and spleen deficiency can be established by CUMS method in late pregnancy.Xiaoyaosan can regulate depression like behavior by changing the signal pathway of PKA/CREB/BDNF mediated by ER?,ER?and GPER in hypothalamus through HPG axis up regulation.4.Based on TCM theories such as 'syndrome detecting from prescription used' and'syndrome differentiation by microscopic method',the syndrome of stagnation of liver qi and spleen deficiency is an important syndrome characteristic of PPD.The biological basis of PPD syndrome of liver stagnation and spleen deficiency with Xiaoyaosan as the evidence is related to the neuroendocrine system,which may be related to the histopathological changes of hypothalamus,the dysfunction of HPG axis,the decrease of sex hormone secretion,the down-regulation of PKA/CREB/BDNF signaling pathway mediated by estrogen receptor in hypothalamus,and the inhibition of neurogenesis.