环磷酰胺对大鼠脾细胞抑制作用的研究  

Studies on immune suppression on spleen cells of rats caused by cyclophosphamide

作　　者：包汇慧[1] 耿雪[1] 贾旭东[1] 汪会玲[1] 支媛[1] 胡静[1] 陈浩[1] 于洲[1] 杨华[1] 

机构地区：[1]卫生部食品安全风险评估重点实验室,国家食品安全风险评估中心,北京100021

出　　处：《毒理学杂志》2014年第3期189-194,共6页Journal of Toxicology

基　　金：公益性行业(农业)科研专项(200903054-06)

摘　　要：目的经口染毒给予大鼠不同剂量环磷酰胺(CTX)以确定脾脏免疫相关指标在大鼠免疫抑制模型中的变化,为毒性评价免疫抑制模型提供科学的检测指标。方法选用40只28日龄Wistar雌性大鼠称重后随机分为对照组及环磷酰胺染毒组(剂量分别为5、10和20 mg/kg·BW)。经口连续染毒环磷酰胺28 d后,取脾脏、胸腺、淋巴结、肝、肾、心经苏木素-伊红(HE)染色后进行形态学观察;FACS法检测脾脏中免疫细胞类型及比例的变化;IHC染色观察免疫细胞形态,FACS法检测脾脏细胞凋亡情况。结果经口给予环磷酰胺28 d后,CTX染毒组大鼠体重显著下降,与对照组比较有统计学意义(P<0.05、P<0.01);5 mg/kg·BW组大鼠脾脏、胸腺指数降低,20 mg/kg·BW组大鼠脾脏指数升高,与对照组比较有统计学意义(P<0.01);组织病理学观察发现脾、胸腺、淋巴结发生不同程度的坏死、凋亡,随着给药剂量的不同,在免疫组织中发生不同程度的纤维结缔组织增生;肝、肾、心脏也发生不同程度的坏死、心肌断裂。脾脏B细胞百分含量随染毒剂量的增加而降低,与对照组比较有统计学意义(P<0.01);5 mg/kg·BW组脾脏T细胞百分含量升高,10和20 mg/kg·BW组脾脏T细胞百分含量降低,与对照组比较有统计学意义(P<0.01);脾脏NK细胞百分含量随染毒剂量升高而升高,10和20 mg/kg·BW组与对照组比较有统计学意义(P<0.01);5 mg/kg·BW组脾脏Th细胞百分含量略有升高,10和20 mg/kg·BW组Th细胞百分含量降低;染毒组脾脏Tc细胞百分含量升高;脾脏细胞凋亡数量随染毒剂量升高而升高,早期凋亡细胞数量与染毒剂量呈正相关性,各染毒组细胞凋亡数量与对照组比较有统计学意义(P<0.01)。结论大鼠CTX 10 mg/kg·BW经口染毒可作为脾细胞免疫抑制模型。CTX通过调节CD8+细胞分化、上调细胞凋亡造成中枢免疫功能低下。Objective In order to verify proper immune indexes of immune suppression of rats for food safety assessment,different doses of cyclophosphamide（ CTX） were given orally to female Wistar rats.Method Forty rats were allocated randomly to four groups（ 0,5,10,20 mg /kg·BW） with ten per group.CTX was orally administrated for 28 days continuously.Spleen,thymus,lymph node,heart,liver and kidney were collected to perform pathological observation; Immune cell types and percentage in spleen were determined by FACS;immune cells’ morphology was showed by IHC staining; cell apoptosis in spleen was tested by FACS.Results Average body weights in CTX exposure groups were decreased significantly compared with those of control group（ P ＆lt; 0.05、P ＆lt; 0.01）.Spleen index and thymus index were decreased in 5 mg /kg·BW group,and were increased in 20 mg /kg·BW group compared with those of control group（ P ＆lt; 0.01）.H.E staining showed that apoptosis and necrosis were found in spleen,thymus,lymph node,different level of fibroplasia in central immune organs was occurred depend on CTX doses.Necrosis was found in liver,kidney and heart,fragmetation of myocardium was also observed in heart.B cell percentage of spleen was decreased along with added CTX doses compared with control group（ P ＆lt; 0.01）; T cell percentage of spleen was increased in 5mg /kgBW and was decreased in 10,20 mg /kg·BW compared with control ones（ P ＆lt; 0.01）; NK cell percentage was increased significantly in 10,20 mg /kg·BW compare with control ones（ P ＆lt; 0.01）.Th cell percentage of spleen was increased slightly in 5 mg /kg·BW and decreased in 10,20 mg /kg·BW; Tc cell percentage of spleen was increased along with added doses.Cell apoptosis was increased in CTX exposure groups compared with control ones（ P ＆lt; 0.01）,and early apoptosis percentage was correlated positively with CTX exposure doses.Conclusion Immune cell types and function were changed along with CTX exposure doses.CTX oral exposure dose should

关 键 词：环磷酰胺 大鼠 免疫抑制 免疫细胞 免疫毒理 

分 类 号：R994[医药卫生—毒理学]