| Objective:Malignant neoplasms are a chronic disease that is a serious threat to human health.So far,malignancy has been a serious disease that affects human health and quality of life.It is the second largest killer of human diseases.Surgery,chemotherapy and radiotherapy are its traditional treatment.Most tumors rely on radiation therapy,but radiotherapy(RT)lacks the selectivity of normal cells and tumor cells,normal cells also exist in the killing effect while killing the tumor cells,resulting in toxic and side effects,making it a great limitation for therapy,and the most common and most critical one is leading to immunosuppression,seriously affecting the patient's treatment and quality of life.Therefore,it is equally important to kill the tumor cells and protect the immune system in the process of tumor treatment.In recent years,the research found that radiation could suppress the immunity on the one hand,on the other hand it could also stimulate the immune function of the body.So immunotherapy is now the most promising strategy to overcome the treatment of cancer,including immunotherapy combined with traditional treatment of malignant tumors.However,the potential immunotoxicity caused by the extensiveness of immunotherapy cannot be negligible,coupled with the high cost of immunomodulating agents and the serious financial burden on patients,so that we are still faced with many challenges.Although traditional Chinese medicine is poor in controlling tumor,most of them have the effect of enhancing immunity,reducing the side effects and are safer,and have unique advantages in radiotherapy adjuvant therapy.Cervus Blood Crystal,a product that is maked by blood of Cervus elaphus or Cervus nipponr after processing and dried,can nourish the blood stasis and increase the energy,release swelling and healing,etc.The aim of this study was to evaluate the effect of Cervus Blood Crystal on the immune response of the tumor-bearing mice with irradiation,by observing the change of the peripheral blood,T cell subsets,and the spleen and bone marrow in histomorphology,to preliminarily explore its possible mechanisms,and lay the foundation for clinical medication..Methods:1 The suspension of S180 tumor cell from ascites was prepared and S180 tumor-bearing mouse model was established.2 Group assignments and administration: Set the normal Group,model group,RT group and RT + Cervus Blood Crystal group.The treatment groups were given X-ray with 10 Gy / 5 f,2Gy / f,once a day for 5 days.The RT + Cervus Blood Crystal group was given the Cervus Blood Crystal suspension via gavage daily for 10 days as the beginning of the irradiation,and the mices were sacried on the 16 th day.3 During the treatment of the mices,monitor the daily state,tumor growth,and regular detection of peripheral blood samples in mice to observe the blood changes of each treatment group,to observe the effect of Cervus Blood Crystal on the daily life quality of mice after irradiation.4 The mices were sacrificed at the end of the treatment,their spleen and ipsilateral femur were collected.The spleen was weighied and the spleen index was calculated.Serum and anticoagulant were prepared.Flow cytometry was used to detect the T Lymphocyte subsets CD3 + T lymphocytes,CD4 + T lymphocytes and CD8 + T lymphocytes.5 The levels of IL-2,IFN-?,IL-4,IL-6 and IL-1 in serum of peripheral blood were measured by enzyme-linked immunosorbent assay(ELISA)and double antibody sandwich method.6 The expression of GATA3 and T-bet in the spleen were detected by immunohistochemistry.7 The histopathological changes of spleen and bone marrow in each treatment group were observed by HE staining technique.Results:1 There was a decrease in body weight of tumor-bearing after radiotherapy,which lost after administration with Cervus Blood Crystal,and tumor growth in tumor-bearing mice was slower than that in RT group.2 The results of peripheral blood test showed that the number of white blood cells,lymphocytes and erythrocytes decreased in 5 days after irradiation compared with the mobel group,Cervus Blood Crystal increased the number of white blood cells and lymphocytes(P<0.05),the most of which was the number of lymphocytes.Especially on the 10 th day,compared with the model group,those of the tumor-bearing mice in RT and RT + Cervus Blood Crystal group were significantly decreased(P<0.01).And the number of lymphocytes on the 10 th and 15 th day in RT + Cervus Blood Crystal group were significantly higher than those in RT group(P<0.05);3 The spleen index: Compared with the model group,the spleen index of the RT model group was decreased,but not statistically significant.Compared with the RT group,the spleen index of the RT + Cervus Blood Crystal group was significantly higher(P<0.01).4 The expression of CD4 + helper T cells,CD8 + killer T cells and CD4 + / CD8 + in the tumor-bearing mice were significantly lower than those in the model group(P<0.05).Compared with the RT group,Cervus Blood Crystal had an enhanced effect on the number of CD4 + helper T cell subsets in peripheral blood of mices after radiotherapy,CD4 + / CD8 + ratio also increased(P<0.01);5 The expression of IFN-? in the peripheral blood of the tumor-bearing mice was decreased(P<0.05),and the level of IFN-? in the peripheral blood of the tumor-bearing mice with Cervus Blood Crystal was higher than that of the RT group(P<0.05).The expression of IL-4 in the peripheral blood of the tumor-bearing mice was decreased after radiotherapy(P<0.05).The expression of IL-6 and IL-1 in the peripheral blood of the RT group was higher than that of the model group,and decreased after the administration.The difference was not significant in statistics(P> 0.05).6 The expression of GATA3 and T-bet in the spleen of the tumor-bearing mice demonstrated that the expression of T-bet in the RT and RT + Cervus Blood Crystal group were both cut down compared with that of the mobel group,but it is shorter in the RT + Cervus Blood Crystal group(P<0.01).Compared with the model group,the expression of T-bet in RT group cut down more than that of GATA3,and the expression of GATA3 was predominant.The expression of GATA3 was significantly decreased(P<0.01)with Cervus Blood Crystal and the expression of T-bet was increased.It was suggested that the reduction of the ratio of T-bet / GATA3 in the spleen tissue of tumor-bearing mices by radiotherapy could be relieved,and the advantage laid with the expression of T-bet.7 The results of histopathological examination suggested that the spleen and bone marrow tissue damages of the tumor-bearing mices with Cervus Blood Crystal were reduced after radiotherapy(P<0.05).Conclusions:Cervus Blood Crystal have a protective effect on injury of blood cells for tumor-bearing mice after radiotherapy,especially T lymphocytes.It can also relieve radiotherapy-induced weight loss and spleen,hematopoietic Functional damage,and balance Th1 / Th2 type cytokines and regulate the CD4 + / CD8 + T cell ratio,which enhance the cellular immune function.Part of the mechanisms may be related to its influence on the expression of T-lymphocyte transcription factors T-bet and GATA3. |
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