硫芥对大鼠淋巴细胞毒性作用的研究  

Study on the effects of sulfur mustard intoxication on lymphocytes in rats

作　　者：杨清武[1] 董兆君[1] 魏相德[1] 张惠丽[2] 

机构地区：[1]第三军医大学预防医学系军事毒理学教研室,重庆400038 [2]解放军北京军医学院军事教研室,北京100710

出　　处：《第三军医大学学报》2001年第1期27-30,共4页Journal of Third Military Medical University

基　　金：全军"九五"指令性课题! (96L0 63 )

摘　　要：目的　探讨硫芥中毒对淋巴细胞的作用机制 ,为寻找硫芥中毒早期改变的敏感指标和提高中毒的防护水平奠定基础。方法　建立硫芥中毒淋巴细胞模型 ,分别用单细胞凝胶电泳法检测淋巴细胞DNA损伤 ;用镀铜镉还原法和一氧化氮合成酶试剂盒检测中毒后淋巴细胞培养上清液一氧化氮 (NO)和一氧化氮合酶 (NOS)的变化 ;用流式细胞仪和琼脂糖凝胶电泳法检测细胞凋亡。结果　硫芥中毒后 4h淋巴细胞DNA明显损伤 ,72h最明显 ,5d开始恢复 ;中毒后NO、NOS的变化相似 ,4h就明显升高 ,1d达到峰值并于 3d开始恢复。NOS的抑制剂 (L NAME)可明显降低NO的含量 ,但对实验组NOS的活性无明显影响。流式细胞仪检测结果显示 4h淋巴细胞凋亡明显 ,而 1、3d几乎无凋亡产生。琼脂糖凝胶电泳结果表明 4h出现典型的细胞凋亡的征象 ,而 1、3d则表现为细胞死亡。结论　 (1)SCG法敏感、经济、简便 ,可用于硫芥中毒细胞毒性、中毒程度的评价 ,在早期诊断上有良好的应用前景 ;(2 )中毒后因NOS的活性升高导致NO的升高而产生细胞毒作用可能是硫芥的中毒机制之一 ;(3 )细胞凋亡参与了硫芥的中毒过程 ,亦可能是其中毒机制之一。Objective To study the effects of sulfur mustard (SM) on lymphocytes and find sensitive parameter for diagnosis at early stage and provide some basis for treatment. Methods The DNA injury was determined with single cell gel electrophoresis, the contents of NO and NOS were measured by NO and NOS kits respectively and apoptosis of lymphocytes was determined by flow cytometry and gelose electrophoresis. Results DNA was obviously damaged after 4h, (DNA migration and percentage of DNA injured lymphocytes were significantly increased) and most prominent at 72 h, began to decrease at 5d. NO and NOS concentration were remarkably increased after 4 h, reached peak at 1 d and recovered since 3 d. Apoptosis of lymphocytes was significantly increased at 4 h, but was not found at 1 d and 3d. DNA LADDER and DNA MADDER were observed. Conclusion (1)SCG is a sensitive, simple and economic method in evaluating the cytotoxicity of SM in toxication and the degree of intoxication, and with good prospects in early diagnasis; (2)The cytotoxicity caused by the elevation of NO induced by the NOS activity after intoxication might be one of the mechanisms of SM intoxication; (3)Apoptosis participates in the process of SM intoxication also might be one of the mechanisms of SM intoxication.

关 键 词：硫芥 淋巴细胞 单细胞凝胶电泳 细胞凋亡 大鼠 

分 类 号：R827.174[医药卫生—临床医学] R392.11