| Because of its high prevalence,high recurrence rate,high disability and high suicidal tendency,depression has become a worldwide public health problem which seriously affecting people’s health.According to the World Health Organization statistics in 2017,as of 2015,depression affected about 322 million people in the world,accounting for 4.4%of the total population,ranking high in the forefront of all diseases.With the accelerating pace of society,depression has gradually become one of the major problems to be solved in China.Breast cancer is a common cancer in women with high incidence rate.The American Cancer Society reports that breast cancer accounts for 30%of all new cases in 2019.One out of every 8 women suffers from breast cancer.It is the most common cancer in women worldwide.At the same time,deaths caused by breast cancer account for 15%of all cancer deaths in women,ranking second.As early as the second Century ad,Galenx,a Greek doctor,observed that depressed women were more likely to have breast cancer than optimistic women,suggesting that depression may be associated with breast cancer.With the development of medicine,more and more scholars adopt the corresponding psychological questionnaire and self rating scale to measure the psychological characteristics of patients with depression and cancer,and explore the impact of depression on the occurrence and prognosis of cancer.At the same time,mitochondria,as the center of energy metabolism in eukaryotic cells,are the important sites of energy metabolism and oxidative stress related reactions such as core carbon metabolism pathway.Brain and tumor are both highly active tissues,which have something in common in high energy demand.Early in the twentieth Century,relevant studies suggested that it was closely related to the development of depression and breast cancer.Related studies have shown that mitochondrial dysfunction can lead to apoptosis of nerve cells,and also participate in the occurrence and development of tumors through immune escape and the formation of tumor inflammatory microenvironment.In the earliest traditional Chinese medical classic "Yellow Emperor Internal Classic",there is a statement that "all kinds of diseases are born in Qi",which has multiple meanings of etiology,pathogenesis and pathogenesis in pathological state.According to traditional Chinese medicine,Qi is an invisible and moving tiny substance.The basic movement form during the period of ascending and descending is the source of body shape,spirit and energy.The ascending and descending movement of Qi and its corresponding changes promote and regulate the functions of various viscera,the metabolism of Qi,blood and body fluid,and the life process of collective growth and aging.The understanding of Qi in TCM coincides with mitochondrial function,thus,we hypothesized that the mitochondrial energy metabolism disorder caused by chronic stress is the biological essence of the pathogenesis of qi deficiency in breast cancer.And qi deficiency is a macroscopic manifestation of mitochondrial energy metabolism disturbance which caused by chronic stress in breast cancer microenvironment,and is an important part in the occurrence and development of breast cancer caused by chronic stress.Traditional Chinese medicine with the function of reinforcing qi and strengthening the liver and relieving stagnation of liver qi can improve the microenvironment of breast cancer by regulating mitochondrial energy metabolism,and then effectively intervene the occurrence and development of breast cancer.Study 1:The correlation between depression and breast cancer riskObjective:To analyze the correlation between depression and breast cancer risk by searching for relevant cohort studies in various databases by means of bioinformatics analysis.Methods:According to the principle of PICOS,the inclusion and exclusion criteria were established,and the quality of the included literature was evaluated by Newcastle Ottawa scale.We retrieved CNKI,Wanfang,VIP,Cochrane Library,PubMed and EMBASE databases comprehensively and systematically.The literature screening and data extraction was independently completed by two researchers.After standardized processing,the data were entered into Microsoft Excel and imported into R Project 4.2.0 for statistical analysis.Chi square test was used to examine the heterogeneity,and I2(i-squared)was used to analyze the heterogeneity.Funnel plot and Egger test were used to assess potential publication bias.At the same time,sensitivity analysis was carried out to eliminate extreme phenomena.All tests were performed by two-tailed test,with P<0.05 as the significant result.Results:(1)After literature screening,a total of 10 papers were finally included in the study,published between 1988 and 2015,mainly from the United States(n=2),Denmark(n=2),the United Kingdom(n=2),South Korea(n=1)and China(n=3).The average score of Newcastle Ottawa scale was 7.5,and the lowest score was 6.The total sample size was 895256,including 273238 females.The study with the least sample size included 3177 people,the most included 601775 people,the study with the least female participants included 472 people,and the most included 87678 people.The included studies were followed up from 5 to 59 years;(2)After the heterogeneity analysis of the included literatures,the random effects model was adopted to fit the results.The final result showed that the risk of breast cancer in the depression group was slightly higher than that in the control group,and there was a weak intensity correlation between depression and breast cancer risk.RR=1.06,95%CI=1.02-1.10);(3)Subgroup analysis of the countries and regions included in the literature showed that:in the subgroup analysis of regions and countries,a significant association was found in Denmark(RR=1.06,95%CI:1.02-1.11);(4)Funnel plot and egger test showed no evidence of publication bias(t=1.18,P=0.2714).Conclusion:Depression is positively correlated with the risk of breast cancer.The risk of breast cancer in depressed people is 1.06 times the risk of breast cancer in those without depression.Study 2:Network pharmacology researchObjective:To explore the main effective components,targets and pathways of XingPiJieYu prescription(XPJY)by network pharmacology,and explore the target and key pathways of breast cancer and depression.The mechanism of XPJY in the prevention and treatment of depression and breast cancer is predicted through the intersection of the target of effective active ingredients and the target of breast cancer and depression,so as to provide reference for further research.Methods:The main active components,target,structure formula and 2D structure diagram of XPJY prescription were obtained from TCMSP and BATMAN database.The obtained structural and 2D structural images were imported into SwissTargetPrediction database.The species was set as "Homo sapiens" for target prediction,and the confidence threshold(probability)≥ 0.2 was used as the evaluation standard for the prediction results to select the prediction targets.Using "breast cancer","breast neoplasm" and "depression" as key words,OMIM,TTD,CTD,Genecards and DisGeNET databases were searched to obtain relevant targets of breast cancer and depression.R Project 4.2.0 was used to obtain the cross targets of active ingredients and diseases,and the Venn diagram was drawn.The intersection target information was imported into Cytoscape 3.8.2 for network analysis,and the network diagram of "traditional Chinese Medicine-active ingredient-action target" was drawn.Using string database,the above intersection targets were imported into the search column,and the species were limited to "Homo sapiens" to construct the protein interaction network.Save the network diagram and related information as TSV format file,import it into Cytoscape 3.8.2 for network topology analysis,and select the core target according to the topology analysis results.The core targets were analyzed by Gene Ontology(GO)and Kyoto Encyclopedia of genes and genomes(KEGG)using metascape database.Results:(1)One hundred and two active components of XPJY formula were obtained and 2448 targets were obtained;(2)Four thousand two hundred and fifty-six breast cancer related targets were obtained,and 3821 with depression.(3)Through topology analysis,we got 37 key targets of breast cancer and XPJY formula,25 key targets of depression and XPJY formula.(4)Two thousand two hundred and eight enrichment results were obtained by GO analysis of the key target of XPJY formula and the breast cancer,2063 enrichment results were obtained by GO analysis of the XPJY and depression key target.(5)KEGG pathway enrichment analysis showed that the key targets of the XPJY and breast cancer were significantly enriched in 116 pathways,which were mainly central carbon metabolism pathway,HIF-1 pathway,cAMP pathway and VEGF pathway.Otherwise,XPJY prescription,the key target of depression,was significantly enriched in 113 pathways,including central carbon metabolism pathway,TNF pathway,PI3K Akt pathway,HIF-1 pathway,etc.Conclusion:XPJY prescription has the characteristics of multi-component,multi-target and multi-pathway synergy,which makes it play a complex role in the treatment of diseases.In this study,we analyzed the common targets and pathways of XPJY prescription,breast cancer and depression through network pharmacology,and predicted the mechanism of XPJY to prevent depression and breast cancer.It may be played by MAPK,mToR,HIF-1,PPARG and other pathways.It reflects the theoretical ideas of "treating the same disease differently","treating different diseases together" and "integrating form and spirit",and provides reference for the follow-up research.Study 3:Mechanism ResearchObjective:Based on SIRT3/PGC-1α To explore the relationship between mitochondrial function and the occurrence and development of breast cancer in depressed rats.Methods:Fifty-two rats were randomly divided into three groups:Depressive breast cancer model group,simple depression group,simple breast cancer group and the blank group,13 rats in each group.(1)The model group of depression breast cancer was induced by chronic unpredictable mild stimulation(CUMS)for 3 weeks,then the sesame oil containing dimethylphenylanthracene(DMBA)was administered to the rats at a 100g/1kg ratio for one time,and continued with CUMS for 12 weeks.(2)After 3 weeks of normal feeding,the breast cancer group received sesame oil containing two methyl benzanthracene(DMBA)and administered with traditional Chinese medicine granules and normal saline for 12 weeks.(3)The simple depression group was given chronic unpredictable mild stimulation for 15 weeks.In the third week,the same volume of sesame oil containing DMBA was given by gavage,and the same volume of normal saline was given by gavage with traditional Chinese medicine granules for 12 weeks.(4)The blank group was fed normally for 3 weeks.In the third week,the same volume of sesame oil containing DMBA was given by gavage,and the same volume of normal saline with traditional Chinese medicine granules was given by gavage for 12 weeks.Result:1.Breast cancer incidence rate and tumor size:Compared with the simple breast cancer group,the incidence and incidence of cancer in the depression breast cancer group increased.2.Tumor volume and quality:Compared with the simple breast cancer group,the volume and quality of the depressive breast cancer group were larger,but there was no significant difference between the two groups.3.Related indexes of mitochondrial function(1)Hippocampus:PGC-lα:Compared with the blank control group,the simple depression group,the simple breast cancer group,the depression breast cancer group PGC-1α expression was increased(P<0.05).SIRT3:Compared with the blank control group,the SIRT3 expression in the simple depression group,the simple breast cancer group and the depression breast cancer group increased(P<0.05).(2)Tumor tissue:PGC-lα:Compared with the depressed breast cancer group,the tumor tissues of the simple breast cancer group and the depressed breast cancer group were PGC-1αProtein expression increased(P<0.05).SIRT3:Negative expression was found in the simple breast cancer group and the depressed breast cancer group.There was no significant difference in SIRT3 expression in each group.(3)Breast tissue:HIF-1a:The expression of HIF-1a in depression breast cancer group was higher than that in other groups,but there was no significant statistical significance.4.Inflammation and immune related indexes(1)Hippocampal:IL-6:Compared with the blank group,the expression of depression group decreased(P<0.05).Compared with the simple depression group,the expression of breast cancer group,depression breast cancer group decreased(P<0.05).TNF-α:Compared with the blank group,the expression of depression breast cancer group decreased(P<0.05).Compared with the simple depression group,the expression of depression group decreased(P<0.05).Compared with the simple breast cancer group,the expression of depression group decreased(P<0.05).IFN-γ:Compared with the depressed breast cancer group,the blank control group,the simple depression group IFN-yexpression was increased(P<0.05).P53:Compared with the depressed breast cancer group,the expression level of P53 increased inother groups(P<0.05).(2)Breast tissue:IL-6:Compared with the blank group,the expression of IL-6 in simple depression group increased(P<0.05);Compared with the depressed breast cancer group,the blank control group,the simple breast cancer group showed lower expression(P<0.05).TNF-α:Compared with the blank group,the expression of the depression group and the depression breast cancer group increased(P<0.05).Compared with the simple depression group,the expression of the depression breast cancer group increased(P<0.05).Compared with depression breast cancer group,the expression of simple breast cancer group were lower and compared with dimple depression group were higher(P<0.05).IFN-y:Compared with the depressed breast cancer group,the other groups increased(P<0.05).P53:Compared with the depressed breast cancer group,the expression level of other groups increased(P<0.05).CD8:Compared with the blank control group,the expression of depression in breast cancer group,depression group,depression group decreased(P<0.05).Compared with depressed breast cancer group,the expression of breast cancer group increased(P<0.01).Compared with the simple breast cancer group,the expression of the depression group,the depression group decreased(P<0.05).NF-κB:Compared with the blank group,expression level of the simple breast cancer group and the depression breast cancer group increased(P<0.05).(3)Tumor tissue:IL-6:Compared with the depressive breast cancer group,the expression of IL-6 in the depression breast cancer group and the simple breast cancer group decreased(P<0.05).TNF-α:In the tumor tissue of rats,compared with the simple breast cancer group,the depression breast cancer group TNF-α expression was increased(P<0.05).IFN-y:Depression,breast cancer,the expression of IFN-y was higher than blank groups,but there was no significant difference among groups.P53:There was no significant difference in the expression of p53 among the groups.CD8:Compared with the depressed breast cancer group,the expression of CD8 in the simple breast cancer group,the depression breast cancer group increased(P<0.05).NF-κB:Compared with the depressed breast cancer group,the other groups NF-κB expression decreased(P<0.05).5.MetabolomicsThe metabolites in the blank group/simple depression group/simple breast cancer group,blank group/simple breast cancer group/depression breast cancer group,simple depression group/simple breast cancer group/depressive breast cancer group were well separated.The pathways involved in different metabolites include arachidonic acid metabolism,phenylalanine metabolism,tryptophan metabolism,pyruvate metabolism,glycerophospholipid metabolism and glycolysis/gluconeogenesis pathway.Pyruvate metabolism,glycerophosphatidylcholine metabolism and glycolysis/gluconeogenesis pathway are closely related to the transition from aerobic phosphorylation to anaerobic glycolysis in tumor tissues.Conclusion:Depression can raise inflammatory factors,promote the formation of tumor microenvironment,and accelerate the occurrence and development of breast cancer.At the same time,depression can also inhibit the recruitment of immune cells around the tumor,reduce the secretion of immune cells,promote the formation of immune escape,and thus accelerate the infiltration and invasion of the tumor.Study 4:Intervention studyObjective:To explore the mechanism of XPJY Recipe on breast cancer in depressed rats based on mitochondrial function.Methods:Fifty-two rats were randomly divided into three groups:Depressive breast cancer model group,depressive breast cancer Chinese medicine group,depression breast cancer group,simple depression group,and simple breast cancer group,13 rats in each group.(1)The model group of depression breast cancer was induced by chronic unpredictable mild stimulation(CUMS)for 3 weeks,then the sesame oil containing dimethylphenylanthracene(DMBA)was administered to the rats at a 100g/1kg ratio for one time,and continued with CUMS for 12 weeks.(2)The rats in the Chinese medicine group with depression and breast cancer were given the model of chronic unpredictable mild stimulation for 3 weeks,and then the sesame oil containing DMBA was added to the CUMS,while the Chinese medicine granules of XPJY was added to the stomach for 12 weeks.(3)In the western medicine group,3 weeks after the chronic unpredictable mild stimulation,the western medicine group was given sesame oil containing DMBA and administered with escitalopram oxalate for 12 weeks at the same time as CUMS.(6)The blank group was fed normally for 3 weeks.In the third week,the same volume of sesame oil containing DMBA was given by gavage,and the same volume of normal saline with traditional Chinese medicine granules was given by gavage for 12 weeks.Result:1.Breast cancer incidence rate and tumor size:The drug intervention group showed a decreasing trend compared with the model group containing cancer,and no significant statistical significance was observed.2.Tumor volume and quality:Compared with the breast cancer depression group,the volume and quality of tumor tissue exfoliated by Chinese medicine group and Western medicine group were smaller than those of the breast cancer depression group(P<0.05).Compared with simple breast cancer group,the volume and quality of tumor tissue in Chinese medicine group with breast cancer depression were smaller(P<0.05).Compared with the depression group,the quality of tumor tissue in the Chinese medicine group was smaller than that in the depression group(P<0.05).3.Related indexes of mitochondrial function(1)Hippocampus:PGC-lα:Compared with the blank control group and the Chinese medicine group,the simple depression group,the simple breast cancer group,the depression breast cancer group PGC-1α The expression was increased(P<0.05):Compared with the traditional Chinese medicine group,PGC-1 in the western medicine group was higher a The expression was increased(P<0.05).SIRT3:Compared with the Chinese medicine group and Western medicine group,the expression of SIRT3 increased in the depressed breast cancer group(P<0.05).(2)Tumor tissue:PGC-lα:There was no significant difference in PGC-1α expression in each group.SIRT3:There was no significant difference in SIRT3 expression in each group.(3)Breast tissue:HIF-la:compared with the depressed breast cancer group,the expression of HIF-1a in the breast tissue of the Chinese medicine group with depression and breast cancer decreased(P<0.05).The incidence of depression breast cancer group was higher than that in other groups,but there was no significant statistical significance.4.Inflammation and immune related indexes(1)Hippocampal:IL-6:Compared with the traditional Chinese medicine group,the western medicine group had lower expression(P<0.05).TNF-α:Compared with the blank group,the expression of depression breast cancer group,Chinese medicine group and Western medicine group decreased(P<0.05).Compared with the traditional Chinese medicine group,the western medicine group had lower expression(P<0.05).IFN-γ:Compared with the depressed breast cancer group,the blank control group,the depression breast cancer Chinese medicine group IFN-yexpression was increased(P<0.05).P53:Compared with the depressed breast cancer group,the expression level of P53 increased in other groups(P<0.05).(2)Breast tissue:IL-6:Compared with the depressed breast cancer group,the blank control group,the Chinese medicine group and the western medicine group showed lower expression(P<0.05).TNF-α:Compared with the blank group,the expression of the depression breast cancer group increased(P<0.05).Compared with the traditional Chinese medicine group,the western medicine group increased the expression(P<0.05).IFN-γ:Compared with the depressed breast cancer group,the other groups increased(P<0.05).P53:compared with the depressed breast cancer group,the expression level of other groups increased(P<0.05).CD8:Compared with the blank control group,the expression of depression breast cancer group and Western medicine group decreased(P<0.05).NF-κB:Compared with the blank group,the Chinese medicine group and the western medicine group,and the depression breast cancer group expression level of NF-κB increased(P<0.05).Compared with the western medicine group,the expression in the traditional Chinese medicine group was lower(P<0.05).(3)Tumor tissue:IL-6:Compared with the depressive breast cancer group,the expression of IL-6 in the depressive breast cancer Chinese medicine group decreased(P<0.05).TNF-α:There was no significant difference in TNF-α expression in each group.IFN-γ:In Chinese medicine group,the expression of IFN-y was higher than that of other groups,but there was no significant difference among groups.P53:The expression of P53 in the Chinese medicine group was higher than that in the other groups.However,there was no significant difference in the expression of p53 among the groups.CD8:Compared with the depressed breast cancer group,the expression of CD8 in the depression breast cancer Chinese medicine group,the Western medicine group increased(P<0.05).NF-κB:Compared with the depressed breast cancer group,the other groups NF-κB expression decreased(P<0.05).Compared with the Chinese medicine group,the western medicine group and the simple breast cancer group NF-κB expression increased(P<0.05).5.MetabolomicsThe metabolites in the depression breast cancer group/Chinese medicine group/Western medicine group,were well separated.The pathways involved in different metabolites include arachidonic acid metabolism,phenylalanine metabolism,tryptophan metabolism,pyruvate metabolism,glycerophospholipid metabolism and glycolysis/gluconeogenesis pathway.Pyruvate metabolism,glycerophosphatidylcholine metabolism and glycolysis/gluconeogenesis pathway are closely related to the transition from aerobic phosphorylation to anaerobic glycolysis in tumor tissues.Conclusion:Based on previous studies,this study found that the antidepressants XPJY recipe and escitalopram oxalate have preventive effects on the occurrence and development of breast cancer with chronic stress depression,and their main mechanisms may be closely related to the inflammatory infiltration,tumor microenvironment and immune escape caused by mitochondrial dysfunction.The effect of escitalopram oxalate is mainly based on improving tumor immune response and inflammatory microenvironment,while XPJY recipe can improve mitochondrial function,inflammatory infiltration,immunosuppression and tumor microenvironment at the same time.Its effect reflects the characteristics of multi-level,multi system and multi-target mechanism of traditional Chinese medicine compound,and reveals the molecular biological mechanism of XPJY recipe in alleviating depressive symptoms and preventing and treating breast cancer. |
