肝微粒体酶对苯诱导的大鼠骨髓细胞凋亡的影响

The Benzene-caused Apoptosis of Bone Marrow Cells of Rats in vitro induced by Microsomal Enzyme of Liver

目的 :观察苯 +S9和单独用苯处理两种方法对体外培养的大鼠骨髓细胞的凋亡效应。方法 :Pereira KI法检测骨髓细胞中髓过氧化物酶的含量 ;荧光染色法检测细胞形态 ;DNA凝胶电泳法观察DNA裂解片段 ;流式细胞仪检测细胞凋亡率。结果 :DNA凝胶电泳 :苯 +S9组苯浓度为 7.5mmol·L-1,诱导时间 1h时即可见模糊的凋亡梯带 ;苯组当诱导浓度为 11.2 5mmol·L-1,诱导时间 3h时 ,电泳图谱呈现典型的DNA梯形条带。流式细胞仪检测各组的凋亡率也表明 ,两组诱导的凋亡呈剂量 效应和时间 效应关系 ,其中苯 +S9组各浓度分组及各时间分组与单独用苯组对应比较 ,凋亡率前者都比后者为高 ,与DNA凝胶电泳的结果相符合。结论 :在体外培养的大鼠骨髓细胞中 ,苯作为一种凋亡诱导剂在S9存在的情况下 ,以更低的剂量和时间点引起大鼠骨髓细胞的凋亡。

Objective: To observe the bone marrow cells(BMC) apoptosis of rats in vitro induced by benzene (groupⅡ)or by benzene+S9 mixture which contained microsomal enzyme of liver (groupⅠ). Methods: The myeloperoxidase(MPO) of BMC was assayed with Pereira-KI method. Fluorescent stain was assayed to study the cells' morphology. DNA splitting fragments were observed by DNA gel electrophoresis. The apoptosis ratio was messured with flow cytometer (FCM). Results: DNA gel electrophoresis showed the benzene-induced final concentration was 7.5 mmol·L -1 at 1 h in group Ⅰ, while the benzene-induced final concentration was 11.25 mmol·L -1 at 3 h in group Ⅱ. And both groups had showed typical DNA ladder. FCM results sbowed both interference (benzene+S9 mixture and benzene sole) had dose effect and time effect on the apoptosis of BMC in vitro. And BMC apoptosis occured early and under lower concentration of benzene in group Ⅰ. Conclusion: Benzene could induce the BMC apoptosis at lower concentration and need less time combined with S9 mixture.