用分子探针杂交法观测多次献血对造血系统的影响

Studying the effect of donation on hematopoietic system by using non-radio-isotopic-label

目的建立非同位素标记的探针杂交测定外周血白细胞端粒DNA长度的方法,借此探讨无偿献血对献血者造血系统的影响。方法酚/氯仿提取基因组DNA,限制性内切酶消化,琼脂糖凝胶电泳,非同位素标记的探针Southern印迹杂交,化学发光X线片曝光显示杂交谱带,积分光密度扫描计算TRF值。结果所测样本获得了较好的低背景杂交谱带,测得35～40岁无偿献血组TRF值平均为11.73kb,相应无献血史对照组平均为11.78kb。结论建立了非同位素标记的探针检测端粒DNA的方法,用上述方法初步显示了固定的长期无偿献血并未对献血者的造血系统产生负面影响。

Objective To measure telomere length by hybridization with non-radio-isotopic-labeled probes, so as to study the effect of donation on hematopoietic system.Methods The high-molecular weight genomic DNA was prepared by standard method and digested, then hybridized by digoxin-labeled probes.The mean telomere length was calculated by scanning the exposed X-ray film with a densitometer. Result The TRF of 35-40 years old donors and nondonors were 11.73Kb and 11.78Kb,respectively. Conclusion A digoxin-labeled and low background method for determining telomere length has been established,which suggested that long-term donation didn＇t affect the hematopoietic system.