摘要
目的探讨碳点(carbon dots,CDs)在MC3T3-E1内的跨膜转运机制与其特点。方法抗坏血酸(ascorbic acid,AA)和聚乙烯亚胺(polyethylenimine,PEI)采用一步微波法制备出荧光CDs。使用MTT法检测CDs对MC3T3-E1增殖的影响,流式细胞术观察MC3T3-E1对CDs的细胞摄取过程,同时观察时间、浓度、温度、跨膜抑制剂及能量抑制剂对CDs跨膜转运的影响。应用原子力显微镜观测在CDs跨膜转运的过程中细胞膜表面形态结构变化。结果 CDs在加入后15min被MC3T3-E1摄取,分布均匀,受时间、浓度、温度和各抑制剂影响,细胞摄取率随时间延长和剂量递增而变化。与对照组相比,低温4℃组能量抑制剂-叠氮化钠(sodium azide,SA)组细胞摄取率显著降低(P<0.01),网格蛋白抑制剂-氯丙嗪(chlorpromazine,CPZ)组细胞摄取率明显降低(P<0.05),无血清培养基(serum-free medium,SFM)组细胞摄取率显著升高(P<0.01)。在细胞摄取15min时细胞膜表面出现大小均匀的凹陷。结论 CDs在MC3T3-E1内的细胞摄取是一个时间、剂量依赖性且耗能的动态过程。血清抑制细胞摄取。CDs主要通过网格蛋白途径进入细胞。
Objective To investigate the transmembrane transport mechanism of carbon dots（CDs） in osteoblast lineage. Methods The ascorbic acid（AA） and polyethylenimine（ PEI ）were used to synthesize the fluorescent carbon dots （CDs）. The effect of CDs on MC3T3-E1 proliferation was examined by MTT. Flow cytometry was used to observe the CDs uptake process. The effects of time points, dose, temperature, transmembrane inhibitors and energy inhibitor on CDs transmembrane were detected. The changes of the plasma membrane were evaluated by atomic force microscope （AFM） imaging. Results CDs uptake happened 15 min after being added and was influenced by time points, dose, temperature and the inhibitors. Compare with the control group, the sodium azide and hlorpromazine cellular uptake rate of 4 ℃ group was lower （ P 〈 0.01）. The cellular uptake rate of serum free medium （SFM） group increased significantly （ P 〈 0.01）. Conclusion The CDs uptake in osteoblast Lineage is a time and dose dependent process. Serum inhibits the cellular uptake. CDs enter the cells mainly through the clathrin pathway.
出处
《北京口腔医学》
CAS
2017年第2期71-75,共5页
Beijing Journal of Stomatology
基金
国家自然科学基金国际(地区)合作与交流项目(8132010801l)
佳木斯大学研究生科技创新项目(YM2016-064)
关键词
碳点
成骨细胞系
跨膜转运机制
Carbon dots
Osteoblast lineage
Transmembrane transport mechanism
