荧光标记对纳米聚苯乙烯细胞毒性和细胞内变化过程的影响

Effects of fluorescent labeling on cytotoxic characteristic and cell distribution of nanopolystyrene

目的探讨荧光标记纳米材料代替未标记纳米材料用于细胞动力学研究的可行性。方法在一定浓度范围(500、250、125、63、32和16μg/ml)和一定时间范围(48、24、16、8、4和1h)内,利用四甲基偶氮唑蓝(MTT)法检测荧光标记与未标记纳米聚苯乙烯对人支气管上皮细胞系BEAS-2B和人单核细胞系THP-1存活率的影响;激光共聚焦显微镜和透射电镜分别观察不同时间内荧光标记与未标记纳米聚苯乙烯在细胞内的分布。结果荧光标记与未标记纳米聚苯乙烯对BEAS-2B和THP-1两种细胞存活率的变化趋势相同,均随着剂量增加细胞存活率降低;而且在相同的剂量组内,荧光标记与未标记纳米聚苯乙烯的细胞存活率差异无显著性(P>0.05);两种纳米材料的细胞存活率剂量-效应曲线趋势和时间-效应曲线趋势也一致;两种纳米材料均可在接触BEAS-2B和THP-1细胞1h后进入细胞,在4h时已经进入细胞核内,在细胞内分布广泛。结论荧光标记对纳米聚苯乙烯细胞毒性和细胞内动力学过程无明显影响,可以代替同尺寸未标记纳米聚苯乙烯用于细胞动力学研究。

Objective To explore the feasibility that studied with fluorescent labeling nano-scale materials cell kinetics.Methods At the doses of 500,250,125,63,32 and 16μg /ml and at the times of 48,24,16,8,4 and 1h,MTT assay was applied to observe the inhibitory effects in BEAS-2B and THP-1 cell lines treated with fluorescent labeling nanopolystyrene and non-labeling nanopolystyrene.Confocal microscopy and transmission electron microscopy were applied to image the translocation of fluorescent labeling nanopolystyrene and non-labeling nano polystyrene into cells.Results The tendency of cell inhibitory rate both in fluorescent labeling nanopolystyrene and non-labeling nanopolystyrene was same,which increase with the dose increase.There were no significant differences in the same dose group for two kinds of materials（P〉 0.05）.The two kinds of materials were found to enter the BEAS-2B and THP-1 on 1 h,and localized within the cell nucleus on 4h.Conclusion There were no obvious influence on cytotoxicity and cell distribution for fluorescent labeling.Fluorescent labeling nanopolystyrene could apply to study cell kinetics equal to the same size nanopolystyrene.