体外培养下氢醌诱导骨髓单个核细胞凋亡的研究

目的　研究体外培养条件下氢醌对正常人骨髓单个核细胞凋亡的影响 ,以探讨苯对造血干细胞的毒性作用。方法　采用HT荧光染色观察形态学变化 ,DNA片段的琼脂糖凝胶电泳检测细胞凋亡 ,流式细胞术ANNEXINV FITC加碘化丙啶 (PI)双染定量检测细胞凋亡率和坏死率的变化。结果　加入不同浓度氢醌培养 6h后出现细胞核荧光强度增强 ,核着色加深 ,核着色不均匀 ,染色质浓集 ,染色质沿核周分布现象。DNA电泳呈典型的“梯形”条带。流式细胞术检测结果发现 ,不同浓度氢醌作用后 ,细胞凋亡率明显高于空白对照组 ,差异有显著性 (P <0 .0 5 )。随着浓度的增加 ,凋亡率明显增加 ,氢醌诱导细胞凋亡的最佳浓度为 5 0 μmol L ,而当氢醌浓度为≥ 75 μmol L时 ,细胞坏死率明显增高。浓度为 5 0 μmol L氢醌诱导的细胞凋亡 ,随着作用时间的延长 ,凋亡率明显增高 ,作用 10h达高峰 ,而后细胞凋亡率下降 ,坏死率增加。结论　体外培养条件下氢醌能诱导骨髓单个核细胞凋亡 ,并存在量 -效与时 -效关系。

Apoptosis induced by hydroquinone in bone marrow mononuclear cells in vitro

OBJECTIVE: To study the effect of hydroquinone on apoptosis of bone marrow mononuclear cells, and to evaluate the toxic effect of benzene on stem cells. METHODS: Cell morphology was observed by HT fluorescent stain method, and DNA fragments were analyzed by agarose gel electrophoresis. Anti-Annexin V FITC plus PI staining for apoptotic and necrotic rate was examined by flow cytometer. RESULTS: After adding different concentrations of hydroquinone to the cells for 6 h culture, the fluorescent intensity of nucleus increased, the color of nucleus became deep and inhomogeneous, and the chromatin was condensed and distributed around the neucleus. DNA ladder was detected in all samples. Cell apoptotic rate in different concentration of hydroquinone groups was significantly higher than that in blank control group (P < 0.05). With the increase of the concentration of hydroquinone, the apoptotic and necrotic rate also increased. The optimal concentration of hydroquinone was 50 micro mol/L. When it was >or= 75 micro mol/L, the necrotic rate increased significantly. Hydroquinone-induced apoptosis was associated with culture time at the concentration of 50 micro mol/L, and the peak apoptotic time was 10 h, then the apoptotic rate decreased and necrotic rate increased. CONCLUSION: Hydroquinone can induce apoptosis of bone marrow mononuclear cells in vitro with dose-effect and time-effect relationship.