| Radiotherapy is one of the main treatment methods for malignant tumors at present,which could kill tumor cells by ionizing radiation.While killing tumor cells,radiotherapy may cause the side effect of bone marrow suppression,and then induce leukopenia.Leukopenia will reduce the patient’s immunity and increase the risk of opportunistic infection,which will affect the continuation of radiotherapy and increase the risk of tumor cell metastasis.Therefore,slowing down the occurrence of leukopenia is the key for tumor radiotherapy to continue.Danggui Buxue Decoction(DBD)was first recorded in the “Neiwaishang Bianhuo Lun”.The whole prescription is composed of Radix Astragali(HQ)and Radix Angelicae Sinensis(DG)in a ratio of 5 : 1.In the prescription,the monarch medicine HQ fortifies “Qi”and invigorates spleen,and the minister medicine DG invigorates blood and promotes blood circulation,and the two drugs are compatible to supplement both “Qi” and blood.Modern pharmacological studies have shown that DBD has many pharmacological effects,such as promoting hematopoiesis,regulating immunity and protecting kidney tissue.Icariin(ICT)is the main active monomer of Epimedii.Our group have found that ICT is one of the key active ingredients in the treatment of cyclophosphamide-induced leukopenia by Qijing Shengbai Granule.In the further study,the efficacy and mechanism of ICT in the treatment of chemotherapy-induced leukopenia were clarified.Therefore,this topic uses DBD and ICT together to form CDBD(Compound Danggui Buxue Decoction)to explore the efficacy and mechanism of CDBD in the treatment of radiation-induced leukopenia.The specific research contents are as follows:1.First,the efficacy of CDBD on radiation-induced leukopenia was verified.The experimental results showed that CDBD could significantly increase the number of white blood cells in peripheral blood,and its effect of increasing white blood cells was better than that of DBD and ICT alone.At the same time,it was found that CDBD could significantly improve the cell viability of bone marrow mononuclear cells,increase the colony forming ability of bone marrow cells,increase the area of bone marrow hematopoietic region,increase the spleen index,reverse the abnormal increase of hematopoietic related factors GM-CSF and IL-6 in serum,and have a clear effect on radiation-induced leukopenia.2.The serum of mice was studied by metabolomics.Based on UPLC-Q-TOF-MS(Ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry)method,a total of 35 endogenous differential metabolites were identified,and CDBD significantly recalled 14 of them.The results of metabolomics showed that CDBD could significantly alleviate the metabolic disorder of irradiated mice.3.The bone marrow cells of mice were studied by transcriptomics.A total of 158 intergroup overlapping differential expressed genes were obtained,and 148 genes were reversely expressed after the administration of CDBD,which significantly alleviated the disorder of gene expression in mice with leukopenia.4.The association analysis was performed on 158 inter-group overlapping differential expressed genes and 35 differential metabolites.The association analysis showed that the genes with higher scores were Jun,Cxcl2 and Egr1,and the metabolites were beta-DGlucuronide,Cholest-5-en-3beta-ol and oxidized glutathione.These key genes and metabolites may be the key regulatory genes and metabolites of CDBD in the treatment of radiation-induced leukopenia.Pathway enrichment analysis showed that the main regulatory pathways of CDBD in raising leukocytes were steroid biosynthesis and glutathione metabolism.Finally,the expression of key genes was verified by qRT-PCR.To sum up,this study is the first to combine traditional Chinese medicine(TCM)-DBD with small molecule-ICT to form a new formula of CDBD,and it has proved that CDBD has good efficacy in radiation-induced leukopenia.This study also preliminarily explored the mechanism of the effect of CDBD on leukemic effect by means of metabolomics and transcriptomics.As a novel TCM with definite curative effect on leukopenia,CDBD is hopeful to be developed into a new clinical drug.The research of this subject will provide a strong scientific basis for the feasibility of the combination of TCM compound and TCM monomer,and also provide more options for the clinical treatment of radiation-induced leukopenia. |
