应用环磷酰胺构建C57BL/6J小鼠免疫抑制模型

背景：在免疫增强剂的研究中,常用到免疫抑制模型,如何建立免疫抑制模型成为免疫增强剂作用评价的关键。目的：应用环磷酰胺构建C57BL/6J小鼠免疫抑制模型。方法：将小鼠随机分为正常对照组、环磷酰胺50 mg/kg 5 d组,环磷酰胺80 mg/kg 3 d组,环磷酰胺80 mg/kg 5 d组,环磷酰胺100 mg/kg 5 d组和环磷酰胺100 mg/kg隔天给药组。正常对照组小鼠腹腔注射生理盐水0.1 mL,连续5 d。环磷酰胺50 mg/kg 5 d组,环磷酰胺80 mg/kg 3 d组,环磷酰胺80 mg/kg 5 d组和环磷酰胺100 mg/kg 5 d组小鼠分别以环磷酰胺50,80,80,100 mg/kg腹腔注射连续5,3,5,5 d。环磷酰胺100 mg/kg隔天给药组小鼠腹腔注射100 mg/kg环磷酰胺,隔天1次,共注射3次。结果与结论：与正常对照组比较,腹腔注射环磷酰胺可导致小鼠外周血中CD3⁺T,CD3⁺CD4⁺T及CD3⁺CD8⁺T细胞明显下降(P < 0. 05);谷丙转氨酶(除环磷酰胺50 mg/kg 5 d、80 mg/kg 3 d组)、谷草转氨酶、尿素显著升高(P < 0.05),其中环磷酰胺80 mg/kg 5 d组、环磷酰胺100 mg/kg 5 d组和环磷酰胺100 mg/kg隔天给药组对肝肾功能的影响更为明显。提示腹腔注射环磷酰胺可建立CD3⁺T,CD3⁺CD4⁺T及CD3⁺CD8⁺T细胞明显下降的免疫抑制模型,其中以环磷酰胺50 mg/kg 5 d和80 mg/kg 3 d方式对肝肾功能的损伤较小。

Establishing a C57BL/6J mouse immunosuppressive model induced by cyclophosphamide

BACKGROUND:The immunosuppressive models are commonly used in the research of immunostimulants, but how to establish the immunosuppressive models is the key for the evaluation of immunostimulants.OBJECTIVE:To establish C57BL/6J mice immunosuppressive model induced by cyclophosphamide.METHODS:Normal C57BL/6J mice were randomly divided into six groups (n=4). Group 1 was normal control group, group 2 was injected with 50 mg/kg cyclophosphamide for 5 days, group 3 was injected with 80 mg/kg cyclophosphamide for 3 days, group 4 was injected with 80 mg/kg cyclophosphamide for 5 days, group 5 was injected with 100 mg/kg cyclophosphamide for 5 days, and group 6 was injected with 100 mg/kg cyclophosphamide every 2 days. Mice in group 1 were intraperitoneal injected with 0.1 mL normal saline and lasted for 5 days. Mice in group 2, 3, 4, 5 were intraperitoneal injected with 50, 80, 80 and 100 mg/kg cyclophosphamide, respectively, and lasted for 5, 3, 5 and 5 days, respectively. Mice in group 6 were intraperitoneal injected with 100 mg/kg cyclophosphamide every two days and total injected for three times.RESULTS AND CONCLUSION:Compared with group 1, the CD3⁺T, CD3⁺CD4⁺T and CD3⁺CD8⁺T levels in the peripheral blood in the other five groups were significantly decreased (P < 0. 05); the contents of alanine aminotransferase (except for group 2 and group 3), aspartate aminotransferase and blood urea nitrogen in the other five groups were significantly increased (P < 0. 05), especially in group 4, 5 and 6. Intraperitoneal injection of cyclophosphamide can establish the immunosuppressive model with decreased level of CD3⁺T, CD3⁺CD4⁺T and CD3⁺CD8⁺T, and intraperitoneal injection of 50 mg/kg and 80 mg/kg cyclophosphamide for 5 and 3 days has less kidney and liver damage.