论文部分内容阅读
目的研究α粒子和NNK联合作用的细胞毒性。方法指数生长期永生化的人支气管上皮细胞(BEP2D细胞)分为正常对照组(NC)、α粒子单纯照射组(α)、NNK染毒组(NNK)、NNK染毒(100μg/ml)后α粒子照射组(NNK+α)和α粒子照射后NNK染毒(100μg/ml)组(α+NNK)。用低密度接种细胞的克隆形成率测定细胞存活分数;用分子探针二氯荧光素双乙酸盐(DCFH-DA)和氢化乙啶(HE)检测细胞内活性氧(ROS)水平;通过测定培养液中乳酸脱氢酶(LDH)活性评价细胞膜通透性损伤。结果与相同剂量NNK或α粒子单独作用比较,α粒子和NNK联合作用BEP2D细胞的存活率明显下降,细胞内ROS水平和细胞培养液中LDH活性显著增高。扣除NNK效应后,α粒子和NNK联合作用BEP2D细胞的存活率明显低于α粒子单独照射组,而细胞内ROS水平和细胞培养液中LDH明显高于α粒子单独照射组。此外还发现α粒子照射后NNK染毒的细胞存活率明显低于NNK染毒后α粒子照射组。结论α粒子合并NNK的细胞毒作用具有协同性,且两者作用顺序不同对细胞存活率有影响。
Aim To study the cytotoxicity of α-particles combined with NNK. Methods Immortalized human bronchial epithelial cells (BEP2D cells) were divided into control group (NC), α-irradiation group (NNK) and NNK group (100μg / ml) (NNK + α) and NNK (100μg / ml) groups after exposure to α particles (α + NNK). Cell viability was determined by clonogenic rate of low-density inoculated cells. Intracellular reactive oxygen species (ROS) levels were detected by molecular probe dichlorofluorescein diacetate (DCFH-DA) and ethidium bromide (HE) Lactate dehydrogenase (LDH) activity in culture medium was evaluated for cell membrane permeability damage. Results Compared with the same dosage of NNK or α particles alone, the survival rate of BEP2D cells decreased significantly with the combination of α particles and NNK, and the intracellular ROS level and LDH activity in cell culture medium were significantly increased. After the NNK effect was deducted, the survival rate of BEP2D cells treated with α particles and NNK was significantly lower than that of α particles alone. The levels of intracellular ROS and LDH in the cell culture medium were significantly higher than those treated with α particles alone. In addition, it was also found that the survival rate of NNK cells exposed to alpha particles was significantly lower than that of alpha particles irradiated with NNK particles. Conclusion The cytotoxicity of α particles combined with NNK is synergistic, and the different order of action of α particles affects the cell survival rate.