蓖麻毒素对原代培养小鼠肺泡上皮细胞的影响  

Effect of ricin on primary culture of murine alveolar epithelial cells

作　　者：王蒙[1,2] 钱军[1,3] 穆成龙[1,2] 赵思言[2,3] 郑关雨[1,2] 付莹莹[1,2] 孙玉成[2,3] 万家余[2,3] 刘文森[2,3] 刘林娜[2,3] 

机构地区：[1]军事医学科学院军事兽医研究所,吉林长春130122 [2]吉林农业大学动物科学技术学院,吉林长春130118 [3]吉林省人兽共患病预防与控制重点实验室,吉林长春130122

出　　处：《中国生物制品学杂志》2013年第8期1113-1116,共4页Chinese Journal of Biologicals

基　　金：国家自然科学基金(31101858)

摘　　要：目的探讨蓖麻毒素气源性中毒中的肺损伤机制。方法取BALB/c雌性小鼠,采用心脏灌注排血、胶原酶和胰蛋白酶原位消化法及组织块细胞迁出培养法,分离培养小鼠肺泡上皮细胞,经刮除法及差相消化贴壁法纯化小鼠肺泡上皮细胞,通过兔抗鼠角蛋白单克隆抗体细胞爬片组化染色法对细胞进行纯度鉴定;经不同终浓度的蓖麻毒素(0、0.000 1、0.001、0.01、0.1、1、10、100μg/ml)处理细胞进行毒性分析;倒置显微镜下观察0.05μg/ml蓖麻毒素对细胞的毒性作用。结果获得的小鼠肺泡上皮细胞纯度达90%以上;随着蓖麻毒素浓度的增加,细胞活力降低,蓖麻毒素浓度与肺泡上皮细胞增殖抑制呈正相关;0.05μg/ml的蓖麻毒素即可引起细胞皱缩、空泡化变性。结论蓖麻毒素可引起原代培养肺泡上皮细胞变性损伤,损伤程度随蓖麻毒素浓度增高而加重,为蓖麻毒素气源性中毒中的肺损伤机制提供了理论依据,同时为进一步探讨蓖麻毒素的毒理作用提供了新的思路。Objective To investigate the mechanism of lung injury caused by ricin aerosol.Methods The alveolar epithelial cells of female BALB / c mice were isolated by cardiac blood ejection through perfusion,in situ digestion with collagenase and trypsin and tissue block migration culture,then purified by curettage and differential adhesion,and determined for purity by histochemical staining of cell slide with anti-cytokeratin monoclonal antibodies.Afterwards,the cells were treated with ricin at various final concentrations（0,0.000 1,0.001,0.01,0.1,1,10 and 100 μg / ml） and analyzed for toxicity.The toxic effect of 0.05 μg / ml ricin on cells was observed under inverted microscope.Results The obtained murine alveolar epithelial cells reached a purity of more than 90%.The cell activity decreased with the increasing concentration of ricin.Ricin showed a dose-dependent inhibitory effect on the proliferation of murine alveolar epithelial cells.The ricin at a concentration of 0.05 μg / ml caused the shrinkage and vacuolated degeneration of the cells.Conclusion Ricin may cause the denatured injury of primary culture of alveolar epithelial cells,in a dosedependent mode.It provides a theoretical basis for study on the mechanism of lung injury caused by ricin aerosol and a new route to further investigation on the toxicological effect of ricin.

关 键 词：蓖麻毒素 肺泡细胞 气源性中毒 培养 

分 类 号：R931.71[医药卫生—生药学]