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生障碍性贫血(aplastic anemia,AA)是一种骨髓造血功能衰竭症,是以全血细胞减少为主要表现的一种综合征[1]。近年来,对AA的机制研究已逐渐深入到分子生物学水平,动物模型的建立是研究其机制的基础。建立动物模型的方法主要包括物理、化学和免疫介导等,很多学者进行了多方面的探索和改进。本研究运用电离辐射及免疫介导在IRM-2小鼠中建立AA小鼠模型,在此基础上对AA小鼠模型的全血细胞、网织红细胞(reticulocyte,Ret)、骨髓及免疫学指标进行了检测,为AA的发病机理、药物筛选及防治研究提供了较好的动物模型。
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AA模型小鼠外周血各指标计数结果见表 1,由表可见,AA模型小鼠的WBC、RBC、HGB含量和血小板数均比对照组和照射组低,其中,与对照组和照射组比较,WBC、RBC和血小板数差异均有统计学意义(WBC:t=27.603、7.835,P均 < 0.05;RBC:t=6.841、6.016,P均 < 0.05;血小板:t=13.944、6.737,P均 < 0.05);与对照组比较,HGB含量差异有统计学意义(t=5.063,P < 0.05)。
组别 只数 WB
(×109/L)RBC
(×1012/L)HGB
(g/L)血小板
(×109/L)对照组 10 8.23±0.41 10.24±0.91 149.7±11.6 1216.4±78.1 照射组 10 4.82±0.83 9.83±0.82 132.1±12.1 996.2±132.1 AA模型组 10 2.38±0.53 8.22±0.21 121.2±13.5 635.1±106.2 注:表中,AA:再生障碍性贫血;HGB:血红蛋白。 表 1 AA模型小鼠外周血各指标计数(x±s)
Table 1. The counts of peripheral blood indexes in aplastic anemia model(x±s)
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AA模型小鼠BMNC数和Ret百分比结果见表 2,由表可见,AA模型小鼠BMNC数和Ret百分比均比对照组和照射组低,且差异有统计学意义(BMNC:t=15.867、2.773,P均 < 0.05;Ret:t=6.245、3.084,P均 < 0.05)。
组别 只数 BMNC(×106/股骨) Ret(‰) 对照组 10 16.5±1.61 76.2±9.7 照射组 10 9.28±2.13 59.8±4.2 AA模型组 10 7.14±1.19 50.2±8.9 注:表中,AA:再生障碍性贫血;BMNC:骨髓有核细胞;Ret:网织红细胞。 表 2 AA模型小鼠骨髓有核细胞数和网织红细胞百分比(x±s)
Table 2. The count of BMNC and percentage of Ret in aplastic anemia model(x±s)
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AA模型小鼠外周血免疫学分型结果见表 3,由表可见,AA模型小鼠CD4+细胞比例低于对照组,差异有统计学意义(t=5.963,P < 0.05);CD8+、CD40+、CD80+/86+细胞比例均高于对照组和照射组,且差异有统计学意义(CD8+:t=8.829、8.048,P均 < 0.05;CD40+:t=10.767、11.072,P均 < 0.05;CD80+/86+:t=6.988、9.241,P均 < 0.05);B220+细胞比例高于对照组,差异有统计学意义(t=2.172,P < 0.05);CD4+/CD8+值低于对照组和照射组,差异有统计学意义(t=5.085、3.718,P均 < 0.05)。
组别 只数 CD4+(%) CD8+(%) B220+(%) CD4+/CD8+值 CD40+(%) CD80+/86+(%) 对照组 10 16.14±3.09 6.83±1.14 33.86±6.39 2.51±0.91 13.33±1.36 21.12±6.06 照射组 10 10.36±6.22 7.14±1.36 41.78±8.26 1.82±0.87 11.26±2.12 15.38±6.35 AA模型组 10 8.91±2.55 13.55±2.12 40.31±6.88 0.73±0.32 20.93±1.77 36.64±3.55 注:表中,AA:再生障碍性贫血。 表 3 AA模型小鼠外周血淋巴细胞亚群比例(x±s)
Table 3. The percentage of lymphocyte subsets in peripheral blood in aplastic anemia model(x±s)
免疫诱导IRM-2小鼠再生障碍性贫血模型的研究
Immune-induced aplastic anemia IRM-2 mice model research
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摘要:
目的 建立IRM-2小鼠再生障碍性贫血(AA)实验动物模型, 并检测其血液及相关免疫学指标。 方法 IRM-2小鼠经137Cs γ射线6 Gy照射后, 输入DBA/2小鼠的胸腺细胞, 建成免疫介导型AA模型。实验分3组, 即对照组、照射组和AA模型组, 于照射后第15天检测小鼠的血常规和免疫学指标。 结果 IRM-2小鼠AA模型组外周血WBC、RBC、骨髓有核细胞数和网织红细胞百分比分别为(2.38±0.53)×109/L、(8.22±0.21)×1012/L、(7.14±1.19)×106/股骨和(50.2±8.9)‰, 均低于对照组[(8.23±0.41)×109/L、(10.24±0.91)×1012/L、(16.5±1.61)×106/股骨和(76.2±9.7)‰]; IRM-2小鼠AA模型组免疫学指标中CD4+细胞比例[(8.91±2.55)%]低于对照组[(16.14±3.09)%], CD8+细胞比例[(13.55±2.12)%]高于对照组[(6.83±1.14)%]。IRM-2小鼠AA模型组外周血及免疫学指标与对照组比较, 差异均具有统计学意义。 结论 IRM-2小鼠AA动物模型的建立, 对AA的研究具有较好的应用价值。 Abstract:Objective To perform traditional blood and immunologic examinations for the pathogenesis of immune-induced aplastic anemia(AA) IRM-2 mice model. Methods IRM-2 mice were divided into three groups, namely, the control, radiation, and AA model groups. IRM-2 inbred mice in the AA model group were used to set up the AA model by whole body 137Cs γ irradiation and eye venous plexus injection with DBA/2 mice thymus cells. After 15 days of radiation, blood and bone marrow cells were counted, and immunologic detection was performed. Results The peripheral WBC, RBC, bone marrow cells, and reticulocyte[(2.38±0.53)×109/L, (8.22±0.21)×1012/L, (7.14±1.19)×106/femur, and(50.2±8.9)‰] in the AA model group were lower than those of the control group [(8.23±0.41)×109/L, (10.24±0.91)×1012/L, (16.5±1.61)×106/femur, and(76.2±9.7)‰]. CD4+ cells [(8.91±2.55)%] of the AA model group were lower than those of the control group[(16.14±3.09)%], whereas CD8+ cells [(13.55±2.12)%] were higher than those of the control group [(6.83±1.14)%]. Peripheral blood and immunological indexes of the AA model group were statistically different from those of the control group. Conclusion IRM-2 mice AA model was successfully established as a good model for the study of AA. -
Key words:
- Anemia, aplastic /
- Models, animal /
- Immune induction /
- Evaluation studies
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表 1 AA模型小鼠外周血各指标计数(x±s)
Table 1. The counts of peripheral blood indexes in aplastic anemia model(x±s)
组别 只数 WB
(×109/L)RBC
(×1012/L)HGB
(g/L)血小板
(×109/L)对照组 10 8.23±0.41 10.24±0.91 149.7±11.6 1216.4±78.1 照射组 10 4.82±0.83 9.83±0.82 132.1±12.1 996.2±132.1 AA模型组 10 2.38±0.53 8.22±0.21 121.2±13.5 635.1±106.2 注:表中,AA:再生障碍性贫血;HGB:血红蛋白。 表 2 AA模型小鼠骨髓有核细胞数和网织红细胞百分比(x±s)
Table 2. The count of BMNC and percentage of Ret in aplastic anemia model(x±s)
组别 只数 BMNC(×106/股骨) Ret(‰) 对照组 10 16.5±1.61 76.2±9.7 照射组 10 9.28±2.13 59.8±4.2 AA模型组 10 7.14±1.19 50.2±8.9 注:表中,AA:再生障碍性贫血;BMNC:骨髓有核细胞;Ret:网织红细胞。 表 3 AA模型小鼠外周血淋巴细胞亚群比例(x±s)
Table 3. The percentage of lymphocyte subsets in peripheral blood in aplastic anemia model(x±s)
组别 只数 CD4+(%) CD8+(%) B220+(%) CD4+/CD8+值 CD40+(%) CD80+/86+(%) 对照组 10 16.14±3.09 6.83±1.14 33.86±6.39 2.51±0.91 13.33±1.36 21.12±6.06 照射组 10 10.36±6.22 7.14±1.36 41.78±8.26 1.82±0.87 11.26±2.12 15.38±6.35 AA模型组 10 8.91±2.55 13.55±2.12 40.31±6.88 0.73±0.32 20.93±1.77 36.64±3.55 注:表中,AA:再生障碍性贫血。 -
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