大麻对正常小鼠脾淋巴细胞的免疫损伤作用  

An study on harmful effect of marijuana on immune in the normal mouse spleen lymphocytes

作　　者：董鹏[1] 卢延旭[1] 张晓宇 穆彪[1] 董佳梓[3] 邓小军[1] 

机构地区：[1]中国刑事警察学院法医学系,辽宁沈阳110035 [2]内蒙古自治区国家安全厅,内蒙古呼和浩特010010 [3]中国中医科学院中医基础理论研究所,北京100700

出　　处：《中国法医学杂志》2010年第5期301-304,共4页Chinese Journal of Forensic Medicine

基　　金：国家"十五"重点科技攻关课题(2001BA801B04)

摘　　要：目的考察大麻对正常小鼠脾细胞的免疫损伤作用。方法采用小鼠脾细胞体外培养法,在细胞培养液中加入100～200μg/mL大麻提取物,连续培养。检测小鼠脾细胞的ConA增殖反应、脾细胞凋亡现象和脾细胞培养上清中IL-2活性;观察脾细胞超微结构的改变和脾细胞内caspase-3活性。结果与正常对照组相比较,大麻染毒组小鼠脾细胞的ConA增殖反应和培养上清中IL-2的活性均显著降低,经Hoechst 33258染色可见大量凋亡的脾细胞;琼脂糖凝胶电泳显示为DNA梯形谱带;透射电镜观察发现100μg/mL大麻组小鼠脾细胞超微结构改变程度随接触时间延长而显著;荧光定量法检测显示100μg/mL大麻染毒组脾细胞内caspase-3活性显著升高。结论大麻提取物能引起小鼠脾淋巴细胞凋亡。Objective To investigate harmful effect of marijuana on immune in the normal mouse spleen lymphocytes.Methods The mouse splenocyte culture method in vitro was adoped.The final concentration of 100/200μg/mL of the marijuana extract were added into the cell culture supernatants,and then continuously cultured.The cell proliferative responses,apoptosis phenomena and the supernatant IL-2 activities were assayed;The splenocyte ultramicrostructure changes and the caspase-3 activity of splenocytes in the 100μg/mL group were observed.Results Compared with those of the normal control group,the ConA-induced splenocyte proliferative reactions and the supernatant IL-2 activities were all reduced markedly,and the large number of apoptotic splenocytes by the Hoechst 33258 stain were showed,the DNA ladder bands were showed by the agarose/ethidium bromide gel electrophoresis in the the 100 and 200μg/mL marijuana extract groups;the ultramicrostructural alteration degree of the splenocytes were more obvious with their longer exposure under transmissible electronic microscope observation,and the increase of caspase-3 activity in the mouse splenocytes was obvious after their exposure to the 100μg/mL marijuana extract by the fluorescence quantitative assay.Conclusion The marijuana extract can cause apoptosis and death of the mouse splenocytes.

关 键 词：法医毒理学 大麻 脾细胞 体外培养 细胞凋亡 半胱氨酸蛋白酶-3 

分 类 号：D919.4[医药卫生—法医学]