Abstract: Objective The aim of present study is to compare the effects of four different modeling method of cyclophosphamide on the immune function of mice. Method Four different ways were employed to develop immunocompromised mouse models by cyclophosphamide, Group A: mice were intraperitoneally injected with cyclophosphamide 80 mg / kg on days 1, 3, and 5, Group B: mice were intraperitoneally injected with cyclophosphamide 60 mg / kg for 5 consecutive days, Group C: mice were intraperitoneally injected with cyclophosphamide 60 mg / kg on days 1, 3, 5, 7, 9, 11 and 13, Group D: mice were intraperitoneally injected with cyclophosphamide 60 mg / kg on days 1, 4, 7, 10 and 13) . The general immunological indexes of mice ( body weight, thymus index, spleen index, whole blood leukocyte and lymphocyte count ) , cellular immunity ( delayed allergic reaction ) and humoral immunity ( determination of half hemolysis value) were observed and compared in the four different models. Result In each experimental group, the relevant detection indexes changed in varying degrees. Compared with the blank control group, the body mass, thymus index, spleen index and WBC count of mice in experimental group A and model group decreased by 19. 15%, 67. 69%, 67. 59% and 83. 87% respectively on the 7th day, while the body mass of mice in model group decreased by 17. 91%, while the thymus index, spleen index and WBC count increased by 7. 14%, 173. 89% and 67. 76% respectively on the 14th day; Compared with the blank control group, the body mass, thymus index and spleen index of mice in experimental group B and model group decreased by 15. 64%, 64. 91% and 64. 60% respectively on the 7th day, and the body mass and thymus index of mice in model group decreased by 17. 86% and 48. 00% respectively on the 14th day, while the spleen index increased by 148. 80%; Compared with the blank control group, the body mass, thymus index, spleen index, WBC count and lymphocyte count of mice in experimental group C and model group decreased by 11. 74%, 82. 61%, 37. 42%, 88. 40% and 91. 44% respectively; Compared with the blank control group, the body mass, thymus index, spleen index, whole blood leukocyte count and lymphocyte count of mice in experimental group D and model group decreased by 10. 83%, 68. 31%, 14. 34%, 66. 15% and 73. 42% respectively. In DTH experiment, the weight difference between left and right ears of immunosuppressive mice decreased by 66. 88%, and in humoral immunity, the serum HC50 of immunosuppressive mice decreased by 74. 52%. In the experiments of group A and B, the relevant detection indexes recovered in varying degrees on the 14th day compared with the 7th day. Meanwhile, the immunosuppressive state of group A mice was more serious in the short term, while the immunosuppressive state of group B mice remained more lasting. Conclusion The above four method could successfully establish immunosuppressive model in mice, and the times, dose, interval days of intraperitoneal injection of cyclophosphamide and the detection endpoint will affect the effect of the immunosuppressive state of mice. So the appropriate model can be selected according to the specific evaluation requirements of drugs.

Key words: cyclophosphamide, immunosuppression, mice

摘要： 目的 研究并对比环磷酰胺 4 种不同造模方式对小鼠免疫功能的影响。 方法 采用 4 种不同方式诱导环磷酰胺小鼠免疫低下模型,A 组:第 1、3、5 天模型组小鼠腹腔内注射环磷酰胺 80 mg / kg;B 组:连续 5 d 模型组小鼠腹腔内注射环磷酰胺 60 mg / kg;C 组:第 1、3、5、7、9、11、13 天模型组小鼠腹腔内注射环磷酰胺 60 mg / kg;D 组:第 1、4、7、10、13 天模型组小鼠腹腔内注射环磷酰胺 60 mg / kg,观察并比较不同造模方式对小鼠一般免疫学指标( 体质量、胸腺指数、脾指数、全血白细胞及淋巴细胞计数) 、细胞免疫(迟发型变态反应) 和体液免疫( 半数溶血值的测定) 等相关指标的影响。 结果 各实验组中模型组的相关检测指标均有不同程度的改变,实验 A 组模型组较空白对照组第 7 天小鼠体质量、胸腺指数、脾指数及全血白细胞计数分别下降 19. 15%、67. 69%、67. 59%、83. 87%,而第 14 天模型组小鼠体质量下降 17. 91%,而胸腺指数、脾指数及全血白细胞计数分别升高了 7. 14%、173. 89%、67. 76%;实验B 组模型组较空白对照组第 7 天小鼠体质量、胸腺指数、脾指数分别下降了 15. 64%、64. 91%、64. 60%,第 14 天模型组小鼠体质量、胸腺指数分别下降 17. 86%、48. 00%,而脾指数升高了 148. 80%;实验 C 组模型组较空白对照组小鼠体质量、胸腺指数、脾指数、全血白细胞计数及淋巴细胞计数分别下降 11. 74%、 82. 61%、 37. 42%、 88. 40%、91. 44%;实验 D 组模型组较空白对照组小鼠体质量、胸腺指数、脾指数、全血白细胞计数及淋巴细胞计数分别下降10. 83%、68. 31%、14. 34%、66. 15%、73. 42%,DTH 实验中,免疫低下小鼠左右耳重量差异减小 66. 88%,体液免疫中,免疫低下小鼠血清 HC50 下降 74. 52%。 对比 A、B 两组,结果显示实验第 14 天相关指标与第 7 天相比均有不同程度的恢复且 A 组小鼠短期内免疫低下状态较为严重,B 组小鼠免疫低下状态维持较持久;延长造模周期时,对比C、D 组结果显示,两种方式均能造成小鼠免疫低下,且 C 组造成的短期内小鼠免疫低下状态较 D 组严重。 结论上述 4 种方法均可成功建立小鼠免疫抑制模型,免疫低下的程度与注射环磷酰胺的次数、剂量、间隔时间及检测终点均有密切关系,可根据具体的药物评价要求,选择合适的评价模型。

关键词: 环磷酰胺, 免疫抑制, 小鼠