大麻对正常小鼠脾淋巴细胞的免疫损伤作用

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。

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

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.