智能姜黄素纳米颗粒联合热疗对肝癌Huh 7细胞的抑制作用

Inhibitory effect of intelligent curcumin nanoparticles combined with hyperthermia on hepatocellular carcinoma Huh 7 cells

目的制备并表征智能姜黄素(curcumin,Cur)-四氧化三铁(Fe_(3)O_(4))纳米颗粒(Cur-Fe_(3)O_(4)NPs),探讨体外协同热疗后对Huh 7细胞的抑制作用。方法采用溶剂热法合成Fe_(3)O_(4)纳米团簇,再化学结合携带Cur,合成Cur-Fe_(3)O_(4)NPs。用傅里叶红外光谱(FT-IR)和透射电子显微镜(TEM)等表征验证。用紫外分光光度法测定NPs的载药率。使用透析法进行药物释放实验,比较射频(radiofrequency,RF)热疗组与无热疗组的Cur释放率。根据不同处理方式将Huh 7细胞分为单纯热疗组、游离Cur组、Cur-Fe_(3)O_(4)NPs组、Cur+RF组和Cur-Fe_(3)O_(4)NPs+RF组,通过CCK-8法评估不同处理组的细胞毒性。结果FT-IR观察到在570 cm^(-1)处是Fe-O键,在963.75 cm^(-1)处Cur连接成功。透射电镜观察Fe_(3)O_(4)纳米团簇和Cur-Fe_(3)O_(4)NPs的粒径分别为(50.7±10.2)nm和(94.0±12.2)nm。Cur-Fe_(3)O_(4)NPs载药率为69.91%。无热疗组Cur-Fe_(3)O_(4)NPs释放Cur约68%,热疗组NPs释放Cur明显增多到85%。细胞毒性实验表明,单纯热疗组和游离Cur组Huh 7细胞存活率均在90%左右;与单纯热疗组和游离Cur组相比,Cur-Fe_(3)O_(4)NPs组细胞活力降低(P<0.05);与其他三组比较,Cur-Fe_(3)O_(4)NPs+RF组Huh 7细胞的存活率明显下降(P<0.0001)。结论Cur-Fe_(3)O_(4)NPs可有效释放Cur,协同热疗后对肝癌Huh 7细胞有明显抑制作用。

Objective To prepare and characterize curcumin(Cur)-Fe_(3)O_(4)nanoparticles(Cur-Fe_(3)O_(4)NPs),and explore the inhibitory effect of NPs plus hyperthermia on the Huh 7 cells in vitro.Methods The Fe_(3)O_(4)nanoclusters were synthesized by solvothermal method,and then chemically bonded with Cur to prepare the Cur-Fe_(3)O_(4)NPs.NPs were verified by Fourier infrared spectroscopy(FT-IR)and transmission electron microscopy(TEM).The encapsulation efficiency of the NPs was determined by ultraviolet spectrophotometry.The drug release experiment in vitro was explored by dialysis method,and the Cur release rate was compared between radiofrequency hyperthermia(RF)group and non-hyperthermia group.According to different treatment methods,the Huh 7 cells were divided into simple hyperthermia group,free Cur group,Cur-Fe_(3)O_(4)NPs group,Cur+RF group and Cur-Fe_(3)O_(4)NPs+RF group,and then the cytotoxicity was evaluated by CCK-8 assay.Results The Fe-O bond was observed at 570 cm^(-1)and the Cur bond was successfully connected at 963.75 cm^(-1)on FT-IR.The TEM images showed that the sizes of Fe_(3)O_(4)nanoclusters and Cur-Fe_(3)O_(4)NPs were(50.7±10.2)nm and(94.0±12.2)nm,respectively.The encapsulation efficiency of NPs was 69.91%.The Cur-Fe_(3)O_(4)NPs responsively released 68%of Cur in non-hyperthermia group,and the Cur release significantly increased to 85%in hyperthermia group.The survival rate of Huh 7 cells in simple hyperthermia group and free Cur group was about 90%.Compared with simple hyperthermia group and free Cur group,the cell viability was decreased in Cur-Fe_(3)O_(4)NPs group(P<0.05).Compared with the other three groups,the survival rate of Huh 7 cells was significantly reduced in Cur-Fe_(3)O_(4)NPs+RF group(P<0.0001).Conclusion The Cur-Fe_(3)O_(4)NPs can responsively release Cur,and significantly inhibit the Huh 7 cells after cooperated with hyperthermia.