一种近红外光线响应的具有抗肝癌活性的一氧化氮纳米载药系统的研究

A near-infrared light-responsive nitric oxide nano-drug carrier system with anti-hepatocellular carcinoma activity

目的探讨一氧化氮(NO)供体和聚多巴胺复合纳米体系的制备方法以及其对肝癌细胞的抑制作用。方法采用化学合成法制备聚多巴胺(PDA)负载一氧化氮供体S-亚硝基半胱胺衍生物(SNO)得到纳米载药复合体系SNO@PDA,通过磁共振成像和质谱分析进行化学结构鉴定,通过动态光散射技术和透射电镜检测其粒径和形貌表征,分别通过电子温度计和Griess试剂盒检测光热稳定性以及NO的释放。最后通过噻唑蓝(MTT)实验检测纳米载药复合体系对肝癌细胞株Huh7的生长抑制作用。两组间均数比较用t检验,多组间均数比较用单因素方差分析。结果磁共振成像和质谱分析确认成功合成SNO,动态光散射技术测定SNO@PDA的粒径在190 nm左右,透射电镜显示为球形颗粒,形态均一,且在生理条件下可稳定存在。SNO@PDA的载药率为19.4%,包封率为71.4%。当浓度为35 mg/L时,近红外(Near-infrared,NIR)照射后温度可升高12℃左右。重复4个开/关循环后升温幅度没有变化。3个循环后NO的释放率达到86%。MTT法测定细胞存活率,当浓度为60 mg/L时,SNO@PDA+NIR组的细胞存活率为(37.2±2.2)%,低于SNO+NIR组(60.1±7.2)%,差异有统计学意义(t=6.500,P<0.01);低于空白材料PDA组(73.9±0.5)%,差异有统计学意义(t=5.900,P<0.01);低于PDA+NIR组(57.9±7.1)%,差异有统计学意义(t=6.400,P<0.01)。结论SNO@PDA可以发挥化疗-光热疗法的组合优势,有效抑制肝癌细胞的增殖。

Objective To investigate the inhibitory effect of the composite nano system of polydopamine and nitric oxide(NO)donor on hepatocellular carcinoma(HCC).Methods A chemical synthesis method was used to prepare polydopamine(PDA)loaded with nitric oxide donor S-nitrosocysteamine derivatives(SNO)to obtain the nano-drug carrier composite system SNO@PDA,and chemical structure was identified by nuclear magnetic resonance(NMR)and mass spectrometry,particle size and morphological characterization were observed by dynamic light scattering technology and transmission electron microscopy,and the photothermal stability and the release of NO were detected by electronic thermometer and Griess kit,respectively.Finally,the methyl thiazolyl tetrazolium(MTT)experiment was used to detect the growth inhibitory effect of the SNO@PDA on the HCC cell line Huh7(purchased from National Collection of Authenticated Cell Cultures).The means between the two groups was compared by t test,and the means among multiple groups was compared by a one-way analysis of variance.Results NMR and mass spectrometry confirmed the successful synthesis of SNO.The particle size of SNO@PDA measured by dynamic light scattering technique was about 190 nm.Transmission electron microscope showed that it was spherical particles with uniform shape,and existed stably under physiological conditions.The drug loading capacity(DLC)of SNO@PDA was 19.4%,and the drug encapsulation efficiency(DEE)of SNO@PDA was 71.4%.When the concentration was 35 mg/L,the temperature could increase by about 12℃ after 6 min of near-infrared(NIR)irradiation.After repeating four on/off cycles,there was no change in the temperature increase range.After three cycles,the release rate of NO reached 86%.The cell viability(%)was determined by the MTT method.When the concentration was 60 mg/L,the cell survival rate in the SNO@PDA+NIR group was(37.2±2.2)%,which was significantly lower than that in the SNO+NIR group[(60.1±7.2)%,t=6.500,P<0.01];significantly lower than in the blank material PDA group[(73.9±0.5)%,t=5.900,P<0.01];significantly lower than in the PDA+NIR group[(57.9±7.1)%,t=6.400,P<0.01].Conclusion The SNO@PDA can exert the synergic chemo-photothermal therapy(CT-PTT)effects,and effectively inhibit the proliferation of Huh7 cells.