氟致牛外周血淋巴细胞凋亡及DNA损伤的效应观察

摘要

目的 探讨氟化钠(NaF)致体外培养的黄牛外周血淋巴细胞损伤和诱导细胞凋亡的作用.方法 应用淋巴细胞分离液分离黄牛外周血淋巴细胞,将不同剂量NaF[0(对照组)、4、8、12、16 mg/L]加入培养体系中,培养24 h后,采用四甲基偶氮唑盐(MTT)比色法检测细胞活性,吖啶橙/溴化乙锭(AO/EB)染色法观察细胞核变化,琼脂糖凝胶电泳法检测DNA断裂情况,碱性单细胞凝胶电泳(SCGE)检测DNA损伤情况.结果 4、8、12、16 mg/L组淋巴细胞的存活率分别为(73.743±0.552)%、(69.184±0.724)%、(65.736±0.055)%、(63.651±0.287)%,较对照组[(100.000±0.003)%]明显下降(P＜0.01).观察到特征性凋亡细胞及有明显的DNA损伤,呈明显的剂量-效应关系(R2=0.7456).结论 一定剂量的NaF有致牛外周血淋巴细胞损伤和凋亡的作用.%Objective To explore the effect of sodium fluoride on DNA damage and apoptosis on ox peripheral blood lymphocyte cultured in vitro.Methods Using lymphoeytes separation medium lymphocytes were separated and different concentration of NaF(0,4,8,12,16 mg/L)were added into the cultual system of lymphocytes for 24 h.Cell viability was measured by MTT.nuclear changes stained by acridine orange-ethidium bromine staining (AO/EB)were observed under fluoroscope,DNA fragment was measured by agarose gel electrophoresis.DNA damage was detected by alkaline SCGE.The differences between each groups were compared.Results Cells were exposed to 4,8,16 and 24 mg/L NaF in 24 h,cell survival rates,respectively being(73.743±0.552)%,(69.184±0.724)%,(65.736±0.055)%and(63.651±0.287)%,decreased significantly compared to control group.There were distinctive cell apoptosis,evident DNA damage and visible dose-effect relation(R2=0.7456).Conclusions A certain concentration of sodium fluoride lcads to lymphocytes apoptosis and DNA damage.