| Objective:Depression is an important factor to promote the development of colorectal tumor,and its mechanism is closely related to immune suppression.Th22 cells are new helper CD4+T lymphocyte subsets,which mainly express cytokine IL-22 to exert its function,by the activation of transcription factor(aryl hydrocarbon receptor,AHR).Early studies of Th22 cells and its cytokine IL-22 mainly focus on inflammation and auto-immune diseases,the knowledge about Th22 cells and IL-22 in blood and colorectal tumor microenvironment of mice bearing colorectal cancer with depression is relatively unclear.In this study,the expression distribution of Th22 cells and related factors IL-22 and to explore its mechanism in mice with colorectal cancer was detected by establishing animal model of colorectal cancer mice with depression and explored the curative effect of antidepressant drug Paroxetine and chemotherapeutic drug Fluorouracil.Methods:The mice were injected with CT-26 colorectal tumor cell which were cultured in vitro subcutaneously in the right armpit of the mouse.After injection,chronic unpredictability mild stress(CUMS)was used to establish the model of colorectal cancer mice with depression.Antidepressant of Paroxetine and chemotherapy of fluorouracil were performed.The BALB/c mice were divided into 8 groups according to the objective of the research.It was the group of normal mice(CON),the group of mice bearing colorectal cancer(CA),the group of mice bearing colorectal cancer with depression(CD),the group of mice bearing colorectal cancer with depression and Paroxetine anti-depressive therapy(CDP),the group of mice bearing colorectal cancer with depression and Fluorouracil chemotherapy(CDF),the group of mice bearing colorectal cancer with depression and Paroxetine combined Fluorouracil therapy(CDP+F),the group of mice with depression(DD),the group of mice bearing colorectal cancer with depression and saline(CDN).Motionless time in forced-swimming test and Sucrose preference test of mice was adopted to evaluate the effect of depression behavior.Measured the weights and the long and short diameter of the tumor of mice once of every three days.The tumor tissues were separated and weighed,and the tumor weight difference was compared between mice of the tumor groups,and the inhibition rate was calculated to evaluate the intervention effect.Assayed the CD3/CD8,CD3/CD4 and Th22 subsets by FCM(flow cytometry),Assayed the level of cytokine IL-22 in the peripheral blood by ELISA.Assayed the level of cytokine IL-22P38? p-p38? ERK?P-ERK,c-JNK and p-JNK in the tumor tissue by western blot.Results:1.Depression and tumor growth of mice.The results of weights of mice showed Group DD had the lower weight than Group CON and CA,Group CD had the lower weight than Group CA,It showed that chronic stress caused the weight loss of DD and CD group mice.The results of consumption of sugar water in Sucrose preference test of mice showed Group DD was lower than Group CON,and Group CD was lower than Group CON and Group CA,Group CD was lower than Group CDP and CDP+F.The results showed that CUMS lead to lack of interest of mice.Paroxetine therapy could reverse the changes of consumption of sugar water in mice of chronic stress,and fluorouracil chemotherapy had no effect.The results of motionless time in forced-swimming test of mice showed Group DD was greater than Group CON,Group CD was greater than Group CA,Group CDP and CDP+F were decreased than Group CD,It showed that chronic unpredictability stress increases the duration of immobility time in mice forced swimming test in Group DD and Group CD,Paroxetine therapy could reverse the changes of immobility time in mice forced swimming test of chronic stress,and fluorouracil chemotherapy had no effect on immobility time of forced swimming test. The results of volume of tumor showed the Group CDF and Group CDP+F had the lower volume of tumor than Group CA?Group CD and Group CDP(26 days?29days and 33 days after tumor injection).The mice of Group CD and Group CDP had the higher volume of tumor than Group CA?Group CDF and Group CDP+F;The mice of Group CD had the highest volume of tumor and the mice of Group CDP+F had the lowest volume of tumor(35 days?38 days and 42 days).The results of weight of tumor showed Group CD had the highest weight of tumor and the mice of Group CDP +F had the lowest weight of tumor.Inhibition rate of tumor showed that Group CDP+F is higher than Group CDF and Group CDP,and Group CDF is higher than Group CDP.There was the highest inhibition rate of tumor in Group CDP+F,It is showed that the Group CDP+F acquired the best treatment effect.It can be seen that the tumor volume of tumor-bearing mice with depressive disorder(Group CD)increased significantly,indicating that depressive disorder had a positive effect on tumor growth.Fluorouracil chemotherapy can inhibit the grow of tumor,Chemotherapy plus Anti-depressive treatment can inhibit the grow of tumor more effectively.2.Flow cytometry analysis for CD4/CD8 of the blood of mice indicated:The results of CD4/CD8 of blood showed Group CD had the lower level of CD4/CD8 than Group CON?Group DD,Group CA had the higher level of Th-22 than Group CON.It is indicated that depression can decrease the percentage of CD4/CD8 in blood of mice with colorectal cancer.The results showed Group CA had the lower level of CD4/CD8 than Group CON and Group CD had the lower level of CD4/CD8 than Group DD.It is indicated that colorectal cancer can make percentage of CD4/CD8 decrease in blood of mice.After treatment,the percentage of CD4/CD8 of Group CD was lower than Group CDF and Group CDP+F,It showed that treatment of paroxetine antidepressant and fluorouracil chemotherapy can increase the level of CD4/CD8.3.Flow cytometry analysis for Th22 of the blood of mice indicated:The results of Th-22 of blood in mice showed Group CD had the higher level of Th-22 than Group CON?Group DD and Group CA,Group CA had the higher level of Th-22 than Group CON.It is indicated that depression can increase content of Th-22 in blood of mice with colorectal cancer,and the colorectal cancer can also make content of Th-22 increase in blood of mice.After treatment,the level of Th-22 cells of Group CDP+F was lower than Group CD,Group CON had lower level of Th-22 than Group CDP?Group CDF and Group CDP+F,Group DD had lower level of Th-22 than Group CDP and Group CDF.It showed that the effect of paroxetine antidepressant treatment and fluorouracil chemotherapy in reducing Th-22 cells was not significant.The combination of paroxetine and fluorouracil therapy can significantly reduce the content of Th-22.After the treatment,The content of Th-22 cell in the blood of mice with colorectal cancer were still higher than the mice of control group.4.ELISA analysis for IL-22 of serum of mice indicated:The results of level of IL-22 in serum showed Group DD had the higher level than Group CON,Group CD had the higher level than Group CON,It showed that depression increased Il-22 level in serum of mice.The results of level of IL-22 in serum showed Group CA had the higher level than Group CON and Group DD,Group CD had the higher level than Group DD.After treatment,the level of IL-22 in serum of Group CD and Group CA were higher than Group CDP? Group CDF and Group CDP+F,the level of IL-22 of Group CA were higher than Group CDP,Group CDP were higher than Group CDF and Group CDP+F,It is indicated that paroxetine antidepressant treatment and fluorouracil chemotherapy can also reduce IL-22 level in serum.The level of IL-22 in serum of Group CD was lower than Group CDP,Group CON was lower than Group CDP?Group CDF and Group CDP+F.It is indicated that the level of IL-22 in serum of mice was still higher than normal and depressed mice after antidepressant and chemotherapy treatment.5.Western blot analysis for IL-22 in the tumor tissue of mice indicated:The results of IL-22 level in tumor showed Group CD had the higher level than Group CA?Group CDP?Group CDF and Group CDP+F,It indicates that depression can make the level of IL-22 increase in tumor tissues of mice,Paroxetine anti-depressive treatment,fluorouracil chemotherapy and combined treatment can drop IL-22 level in the tumor tissue of mice.After treatment,the level of IL-22 in tumor tissues of Group CA was lower than Group CDP ?Group CDF and Group,Group CDP+F was lower than Group CDP.The level of IL-22 in tissues was higher than that of Group CA after treatment with paroxetine,fluorouracil and combined therapy.It can be seen that paroxetine anti-depressant treatment and fluorouracil chemotherapy and anti-depression combined chemotherapy can reduce the level of IL-22 in tumor tissues of mice,and the level of IL-22 in tumor tissues of mice was still higher than mice with colorectal cancer after antidepressant and chemotherapy treatment.6.Western blot analysis for p-p38?p-ERK?p-JNK in the tumor tissue of mice indicated:The results of p-p38?p-ERK?p-JNK level in tumor showed Group CD had the higher level than Group CA?Group CDP?Group CDF and Group CDP+F,It indicates that depression can make the level of p-p38?p-ERK?p-JNK increase in tumor tissues of mice,Paroxetine anti-depressive treatment,fluorouracil chemotherapy and combined treatment can drop p-p38?p-ERK?p-JNK level in the tumor tissue of mice.After treatment,the level of p-p38?p-ERK and p-JNK in tissues of Group CD was higher than that of Group CDP?Group CDF and Group CDP+F.and p-p38,p-ERK and p-JNK proteins were significantly reduced in Group CDP+F.It can be seen that paroxetine anti-depressant treatment and fluorouracil chemotherapy and anti-depression combined chemotherapy can reduce the level of p-p38?p-ERK and p-JNK in tumor tissues of mice.Conclusions : Chronic mild stress could reduce the body weight and prolonged motionless time in forced-swimming test of mice with colorectal tumor,the depression could make the weight of tumor increase and induce grow of colorectal tumor.Paroxetine and chemotherapy could inhibit grow of the tumor.The amount of Th-22(%)and IL-22 in blood of mice with colorectal tumor were higher than the other groups,IL-22 in tissue of mice with colorectal tumor were higher than the other groups,Paroxetine with Fluorouracil treatment could reduce the level of Th-22(%)and IL-22 effectively,possibly through inhibiting IL-22 expression to regulate the activity of the mitogen-activated protein kinase(MAPK)signaling pathway. |
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