硫芥对大鼠脾脏淋巴细胞线粒体的损伤作用

The damaging effect of sulfur mustard on the mitochondria in rat lymphocytes in vitro

目的观察硫芥对大鼠脾脏淋巴细胞线粒体的损伤作用。方法用密度梯度离心法分离大鼠脾淋巴细胞,与硫芥共同进行体外培养。用琼脂糖DNA凝胶电泳观察细胞凋亡的发生;用W estern b lot观察Cyt-c释放。用MTT法检测线粒体功能状态,以罗丹明123标记的荧光探针检测线粒体跨膜电位。结果100μmol/L硫芥作用4 h线粒体功能就有降低,线粒体跨膜电位降低,二者间呈直线相关。硫芥中毒早期即可引起淋巴细胞线粒体细胞色素C释放和细胞凋亡(DNA ladder)。结论硫芥可引起大鼠脾淋巴细胞线粒体明显损伤,线粒体参与了硫芥的细胞毒性作用过程。

Objective To investigate the damaging effect of sulfur mustard on the mitochondria in rat lymphocytes in vitro. Methods Rat spleen lymphocytes were isolated by density gradient centrifugation and cultured with 100μmol/L sulfur mustard in vitro, DNA ladderring was used to detect the cell apoptosis. Cyt-c release was measured by Western blotting. The mitochondria function was detected by MTT method. Fluorescent probe labeled with Rhodamine 123 was used to study mitochondria potential. Results The mitochondria potential and cell viability decreased at 4 h after treated with 100 μmol/L sulfur mustard. There was a liner correlation between the mitochondria potential and cell viability in lymphocytes. The content of Cyt-c increased significantly at 4 h as compared with control group in Western blotting. Conclusion Sulfur mustard could damage the rat lymphocyte mitochondria significantly, and mitochondria participates in cytotoxic effect induced by the sulfur mustard.