安徽医科大学学报2025，Vol.60Issue(8)：1387-1393,7.DOI:10.19405/j.cnki.issn1000-1492.2025.08.004
不同剂量X射线联合异体淋巴细胞输注对建立再生障碍性贫血小鼠模型的影响
Influence of different doses of X-ray irradiation combined with allogeneic lymphocyte infusion to establish a mouse model of aplastic anemia
摘要
Abstract
Objective To investigate the effects of different doses of X-rays irradiation combined with allogeneic lymphocyte infusion on the establishment of aplastic anemia in mice.Methods Forty BALB/c mice were randomly divided into four groups:the 3 Gy group(n=9),the 4 Gy group(n=9),the 5 Gy group(n=10),and the con-trol group(n=12).In the 3 Gy,4 Gy,and 5 Gy groups,the experimental mice were exposed to corresponding do-ses of X-ray and then intravenously infused with 0.2 mL mixed suspension of the thymus and spleen cells from DBA/2 mice,at a concentration of 1 × 107 cells/mL,within 4 hours after irradiation.The control group did not un-dergo X-ray irradiation and infused with an equivalent volume of physiological saline instead.Blood samples were collected from the orbital venous plexus of BALB/c mice and analyzed using an animal automated hematology analy-zer to measure peripheral blood parameters,including red blood cells(RBC),white blood cells(WBC),and platelets(PLT).The general condition of mice was monitored daily,and survival rates were recorded for each group.At the experimental endpoint,the tibias were harvested for hematoxylin and eosin(HE)staining,while the femurs were used to prepare bone marrow smears for morphological examination.For the 5 Gy group,T-cell subsets were analyzed by flow cytometry,and cytokine levels were quantified using enzyme-linked immunosorbent assay(ELISA)at the endpoint.Results In the 3 Gy group,pancytopenia was observed,but platelet recovery occured rapidly,returning to normal levels by day 17 post-modeling.No deaths occurred during the observation period.At the experimental endpoint,bone marrow morphological examination revealed hypercellular marrow with a normal myeloid-to-erythroid(M/E)ratio,and no significant morphological abnormalities were noted in cells at any devel-opmental stage.HE staining demonstrated that the control group exhibited densely packed bone marrow tissue filled with hematopoietic cells.In the 4 Gy group,pancytopenia persisted throughout the observation period.The survival rate was 90％.Endpoint analysis showed hypocellular marrow by morphological examination.HE staining indicated minimal fatty infiltration in the bone marrow tissue.In the 5 Gy group,pancytopenia was observed,though erythro-cyte counts returned to normal levels by day 24.The survival rate during the observation period was 50％.End-point analysis revealed vacuolization of marrow particles and reduced hematopoietic cells with predominantly non-hematopoietic cells in bone marrow morphology.HE staining demonstrated severe fatty infiltration in the bone mar-row tissue,with scarcity of immature cells and hematopoietic precursor cells.Flow cytometry analysis showed a de-creased proportion of CD4+T cells(％)and an increased proportion of CD8+T cells(％).ELISA confirmed ele-vated secretion of negative hematopoietic regulators:interferon-gamma(IFN-γ)and tumor necrosis factor-alpha(TNF-α).Conclusion Combined administration of varying radiation doses with allogeneic lymphocyte infusion consistently induced peripheral blood cytopenia in mice,characterized by reductions in RBC,WBC,and PLT counts.Integrated analysis of bone marrow morphology,histopathological assessment via HE staining,and immuno-logical parameters confirmed that a mouse model of aplastic anemia can be successfully established using 5 Gy X-ray irradiation coupled with infusion of 2 × 106 allogeneic lymphocytes.关键词
再生障碍性贫血/异体淋巴细胞/X射线/外周血/生存率/骨髓/免疫Key words
aplastic anemia/allogeneic lymphocyte/X-ray/peripheral blood/survival rate/bone marrow/immunity分类
医药卫生引用本文复制引用
周晓晓,徐彩仙,汪桂琴,洪强,陶千山,李丛,王会平,翟志敏..不同剂量X射线联合异体淋巴细胞输注对建立再生障碍性贫血小鼠模型的影响[J].安徽医科大学学报,2025,60(8):1387-1393,7.基金项目
国家自然科学基金面上项目(编号:82370225) （编号:82370225）
安徽省转化医学研究院科研基金项目(编号:2023zhyx-B08) National Natural Science Foundation of China General Program(No.82370225) （编号:2023zhyx-B08）
Research Pro-ject of Anhui Provincial Institute of Translational Medicine(No.2023zhyx-B08) （No.2023zhyx-B08）
