| Objectives1 To provide guidance and new ideas for clinical diagnosis and treatment of PCOS,we systematically consulted and summarized relevant literature on polycystic ovary syndrome(PCOS)from home and abroad,and elaborated the relevance between PCOS and psychological stress.2 The characteristic of TCM syndrome distribution of PCOS was analyzed by cross-sectional study.By the pointcut of chronic psychological stress,this paper analyzes the correlation between life quality,psychological state and personality characteristics and TCM syndromes,and discusses the relationship between TCM syndrome types of PCOS and chronic psychological stress,clarifies the importance of "kidney deficiency and liver stagnation" in PCOS pathogenesis,and provides a theoretical basis for TCM syndrome differentiation and treatment.3 Testify the important effects of chronic psychological stress on PCOS,basing on rat model established by letrozole and chronic mild and unpredictable stress(CUMS)method.Take this model as the study object,combined with reproductive and psychological stress indicators,discussing the therapeutic effect and mechanism on rats of Bushen Jieyu Tiaochong Decoction.The ovarian granule cell apoptosis pathway of PERK-ATF4-CHOP induced by endoplasmic reticulum stress was used as a new target to investigate the therapeutic mechanism of Bushen Jieyu Tiaochong Decoction on PCOS in rats with chronic stress.Methods1 Clinical researchThe clinical information of 309 patients with PCOS was collected and sorted.The patients'quality of life,the changes of anxiety and depression,and the personality characteristics were investigated by the health survey summary(SF-36),the anxiety/depression self-rating scale(SAS/SDS),and the Eysenck personality questionnaire,Chinese version(EPQ-RSC).Analyze the TCM syndrome characteristics of PCOS patients and their differences in quality of life,mental health and personality characteristics with different syndrome types.Investigate the dependency relationship between pathogenic factors,anxiety and depression,and personality characteristics in PCOS patients of kidney deficiency and liver depression syndrome by logistic regression model.2 Experimental researchUse CUMS method to simulate chronic psychological stress.Use letrozole to build PCOS rat model.Rats were randomly divided into blank group,chronic stress group(CUMS),PCOS group,and PCOS combined chronic stress group(PCOS+CUMS).The model was compared and evaluated with the indexes of estrous cycle,behavior,ovarian histopathology,serum sex hormones and neurotransmitters in hippocampal tissue.Use PCOS+CUMS animal model to study the therapeutic mechanism.In addition to the normal group,the model rats were randomly divided into the model group,the Diane-35 group,the simple Bushen group(i.e.,Bushen group),and the low-dose,medium-dose and high-dose of Bushen Jieyu Tiaochong groups(i.e.,Bushen Jieyu group).Behavioral tests were carried out,as well as the measurement of the ovarian volume and ovarian quality of rats in each group.HE staining was used to observe the pathological morphology of ovarian tissue.Serum levels of LH,FSH and FT were tested by ELISA.The levels of NE,5-HT and 5-HIAA in hippocampus were tested by HPLC-ECD.Choose the group with better efficacy among the Bushen Jieyu groups of low,medium and high dose,as well as the rest of other groups,to study the therapeutic mechanism.Tunel method was used to detect the apoptosis of ovarian granule cells.Immunohistochemistry was used to detect the expression and localization of GRP78 and CHOP proteins in ovarian tissues.The expression levels of CHOP,GRP78,PERK and ATF4 proteins in ovarian tissues were detected by Western Blot.Results1 Clinical research(1)There were 309 effective cases in this study.The proportion of patients aged 27-33 years old was the highest(55.34%).Junior college and above degrees take up the major part(91.26%),and the patients are mostly mental workers(75.4%).The proportion of obese patients was 36.89%,67.96%of patients had no exercising habits,the proportion of patients with menstrual disorders was 95.47%,the proportion of patients with a history of infertility was 29.13%,and primary infertility was accounted for 64.44%in infertility patients,while secondary accounted for 35.56%.(2)Among the 309 patients with PCOS,kidney deficiency and liver stagnation syndrome accounted for the highest proportion(37.22%),followed by phlegm-stasis syndrome(23.30%),phlegm-dampness syndrome(20.39%)and kidney deficiency and blood stasis syndrome(19.09%).(3)There were differences in education level,job nature and BMI among different syndrome types(p<0.05).The proportion of patients with junior college and above degrees was the highest in kidney deficiency and liver depression syndrome(95.65%).The proportion of mental worker patients is high in kidney deficiency and liver stagnation syndrome,as well as spleen deficiency and phlegm and dampness syndrome,which is 80.95%and 80.87%respectively.The BMI of PCOS patients of spleen deficiency and phlegm and dampness syndrome and phlegm-stasis syndrome were higher than that of kidney deficiency and liver stagnation syndrome,as well as kidney deficiency and blood stasis syndrome(p<0.05).(4)Overall health(GH),emotional function(RE),mental health(MH),and vitality(VT)scores were lower in SF-36.Those with anxiety and depression status accounted for 28.8%and 40.13%each in the total number of PCOS patients.(5)There were statistical significance difference in VT and MH among different TCM syndromes(p<0.05).The scores of VT dimension and MH dimension of kidney deficiency and liver stagnation syndrome were lower than those of spleen deficiency and phlegm and dampness syndrome(p<0.05).The total score of SF-36 mental health in PCOS patients with kidney deficiency and liver depression syndrome is lower than the other three syndromes,and SAS and SDS were higher(p<0.05).(6)There were statistical significance in E,N and P scores in EPQ-RSC among TCM syndromes(p<0.05).The E dimension score of spleen deficiency and phlegm dampness syndrome was the highest,and the kidney deficiency and liver depression syndrome was the lowest.The N and P scores of kidney deficiency and liver depression were the highest,and the lowest was spleen deficiency and phlegm dampness syndrome.(7)Further Logistic regression analysis was carried out on the kidney deficiency and liver stagnation syndrome,which took up the highest proportion of TCM syndrome types.Age,education level,job nature,BMI,depression and neuroticism N were related to the syndrome of kidney deficiency and liver stagnation(p<0.05).Younger age(take 34-40 years old for reference),lower weight(take overweight and obesity for reference),and depressionwere positively correlated with kidney deficiency and liver stagnation syndrome.High school and technical secondary school education(take college degree or above for reference),labor work(take the unemployed for reference),emotional stability or personality neurotic intermediate type(take the emotional instability for reference)were negatively correlated with kidney deficiency and liver depression syndrome.2 Experiment 1Body mass,serum FT level and LH/FSH ratio of rats in PCOS group and PCOS+CUMS group were higher than those in the blank group(p<0.05).PCOS+CUMS group had more obvious polycystic changes in ovarian tissue than PCOS group.In the CUMS group and the PCOS+CUMS group,the horizontal movement score,vertical movement score,sucrose bias and NE and 5-HT contents in the hippocampus were lower than those in the blank group(p<0.05),and the contents of 5-HIAA and the ratio of 5-HIAA/5-HT were higher than those in the blank group(p<0.05).3 Experiment 2(1)Body mass,ovarian volume and ovarian mass:the body mass,left ovarian volume and ovarian mass of rats in the model group increased significantly compared with that in the normal group(p<0.05).The body mass of rats in Diane-35 group,Bushen group and medium dose of Bushen Jieyu group was obviously lower compared with model group(p<0.05).The left side of the ovaries volume and quality of rats from medium dose of Bushen Jieyu group were obviously decreased(p<0.05);(2)Ovarian histopathology:multiple cystic dilated follicles were observed in the model group,with loose arrangement of granule cells and reduced number of layers.Normal follicles and corpus luteum were observed in the medium and high dose of Bushen Jieyu groups,while the follicles with cystic dilatation were rare.The granule cells were arranged in a regular manner,and the number of layers increased compared with the model group.(3)Sex hormones:the levels of LH and FT and the ratio of LH/FSH in the model group were higher than that in the normal group(p<0.05).Compared with model group,the FT level significantly decreased in Diane-35 and medium dose of Bushen Jieyu group(p<0.05).The level of LH,LH/FSH ratio in Diane-35 and the medium and high dose of Bushen Jieyu groups were significantly decreased(p<0.05).The level of LH in the Bushen group was also lower than that in the model group(p<0.05).4 Experiment 3(1)Behavior:the scores of horizontal and vertical movement and the degree of sucrose bias in the model group were lower than those in the normal group(p<0.05).Compared with the model group,the horizontal movement score of the Bushen group,as well as the medium dose of Bushen Jieyu group were significantly increased(p<0.05).Although there was no significant difference in the vertical movement score and sucrose eccentricity between each treatment groups(p>0.05),the vertical movement score of the rats in the medium dose of Bushen Jieyu group and Bushen group showed an increasing trend,comparing with that before treatment.The increase of sucrose eccentricity in the medium dose of Bushen Jieyu group was the largest.(2)Neurotransmitters:the levels of NE and 5-HT in the model group were lower and the normal group,while 5-HIAA were higher than those in the normal group(p<0.05),the ratio of 5-HIAA/5-HT was higher,but there was no statistical significance(p>0.05).Compared with model group,content of NE was significantly increased in Bushen group and each dose of Bushen Jieyu group(p<0.05),the 5-HT,5-HIAA and 5-HIAA/5-HT ratio of each treatment group had no significant differences(p>0.05).5-HT was the highest in medium dose of Bushen Jieyu group,and 5-HIAA was the lowest in high dose of Bushen Jieyu group;5 Experiment 4(1)Granule cell apoptosis:compared with the normal group,the apoptosis index of follicular granule cell in the model group was significantly higher(p<0.05).Compared with the model group,the apoptosis index was significantly reduced in the Diane-35 group and the medium dose of Bushen Jieyu group(p<0.05).(2)Expressions of GRP78,CHOP,PERK and ATF4 proteins:GRP78 and CHOP proteins were mainly expressed in the granule cell layer in the ovarian tissues of the model rats group.Compared with the normal group,the expressions of GRP78,CHOP and ATF4 proteins in the model group were significantly up-regulated(p<0.05),and the expression level of PERK protein in the model group was increased,but there's no statistically significant difference(p>0.05).Compared with model group,GRP78,ATF4 protein expression in Diane-35 group,Bushen group and medium dose of Bushen Jieyu group were significantly decreased(p<0.05).CHOP protein expression in medium dose of Bushen Jieyu group was significantly decreased(p<0.05),and the PERK protein expression levels dropped in the each treatment group,but there was no statistically significant difference(p>0.05).Conclusions1 According to the literature review,PCOS is an endocrine disorder disease with reproductive dysfunction and metabolic disorder,and psychological stress plays an important role in the occurrence and development of PCOS.Traditional Chinese medicine has accumulated rich experience in the treatment of PCOS,so the mechanism and advantages of traditional Chinese medicine in the treatment of PCOS should be further studied in combination with psychological stress factors.2 Clinical studies have confirmed that kidney deficiency and liver stagnation is a common TCM syndrome of PCOS.PCOS patients with kidney deficiency and liver stagnation suffer from greater mental pressure and psychological burden,and are more likely to have introversion,emotional instability,anxiety and tension and other personality tendencies."kidney deficiency and liver stagnation" is closely related to the chronic psychological stress state of PCOS patients.Patients with younger age,lighter weight,depression and unstable mood are more likely to be diagnosed as kidney deficiency and liver stagnation syndrome.3 PCOS combined chronic stress state animal model,built by letrozole and CUMS,is closer to the clinical practice in ovarian pathological changes,endocrine disorder,behavioral changes and neurotransmitters,which can be used as a reliable animal model for the study of PCOS combined pathological stress and pharmacodynamic mechanism.And the study further confirmed that the chronic psychological stress and PCOS are influential to each other.4 Bushen Jieyu Tiaochong Decoction can promote the restoration of balance of reproductive endocrine environment in model rats,restore follicular development and ovulation,and improve the behavioral performance of model rats.The therapeutic mechanism of this decoction might be related to the regulation of monoamine neurotransmitters in the brain of model rats,and the inhibition of PERK-ATF4-CHOP signaling pathway,down-regulating GRP78 expression to further delay the ovarian granule cell apoptosis mediating by ERS. |
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