硫芥诱导大鼠小肠上皮细胞凋亡的研究

目的：观察硫茶对大鼠小肠上皮细胞（IEC－6）生长的影响及其引起的细胞死亡的方式。方法：透射电镜、DNA琼脂糖凝胶电泳定性硫芥中毒的IEC－6细胞形态变化及DNA的改变。采用Cell Death DetectionELISA试剂盒研究IEC－6细胞凋亡与硫芥浓度之间的关系。结果：0．1－1000μmol/L硫芥对IEC－6细胞的毒性作用与硫芥的深度和作用时间呈正相关。电镜观察到细胞凋亡的形态学改变。细胞膜完整,染色质固缩并聚集于核膜边缘,凋亡小体形态;DNA电泳有“DNA Ladder”出现,0．1－100μmol/L硫芥处理的细胞的凋亡量随硫芥 度的升高而增加,超过100μmol/L测凋亡量反降低。结论：硫芥可诱导大鼠小肠上皮IEC－6细胞凋亡,在低浓度时以凋亡为主,高浓度时则出现坏死。

Apoptosis induced by sulfur mustard in the intestinal epithelioid cell line 6 in rats

Objective： To observe the mode of cell death induced by sulfurmustard in the intestinal epithelioid cell line 6 (IEC-6) in rats. Methods：IEC-6 cells were incubated in vitro with a wide range concentration of sulfur mustard (0.1-1 000 μmol/L). Cell morphological changes were observed by transmissive electron-microscope, and DNA fragmentations were detected by agarose gel electrophoresis. The relationship between the concentration of sulfur mustard and apoptosis of IEC-6 cells was measured by Cell Death Detection ELISA kit. Results：Sulfur mustard-injured (0.1-1 000 μmol/L) IEC-6 cells showed a positive correlation between the culture period and concentrations of sulfur mustard, and also showed typical morphological characteristic of apoptosis such as intact cell membrane, chromatin condensation and apoptotic body formation. The detection of DNA electrophoresis presented “DNA Ladder” which was considered the hallmark of apoptosis. Quantitatively measured by Cell Death Detection ELISA kit, it was found that apoptosis of IEC-6 cells increased with the increasing concentration of sulfur mustard and peaked at 100 μmol/L. Exceeding this concentration apoptosis declined.Conclusion：Sulfur mustard at low concentration causes IEC-6 cells apoptosis, whereas at high concentration induces necrosis.