紫杉醇纳米粒的制备及其对肝癌HepG2和Huh-7细胞的影响

Preparation of paclitaxel-loaded nanoparticles and their effect on hepatocellular carcinoma HepG2 and Huh-7 cells

目的探讨具有生物活性并可控制释放的紫杉醇纳米粒的制备及其对肝癌HepG2和Huh-7细胞的作用。方法应用单乳溶媒挥发法制备聚乳酸-聚羟基乙酸共聚物装载的紫杉醇纳米粒。高效液相色谱分析纳米粒的包封率、载药率和模拟体外药物释放率,扫描电子显微镜观察纳米粒的形态,粒度分析仪检测纳米粒粒径。肝癌HepG2和Huh-7细胞与紫杉醇纳米粒共培养,通过倒置显微镜观察细胞形态变化,采用四甲基偶氮唑盐(MTT)法检测细胞存活率。结果紫杉醇纳米粒呈光滑球形,平均粒径216 nm,包封率87.4%,载药率1.79%,模拟体外释药曲线示早期呈爆发释放,随后为缓慢持续释放。紫杉醇纳米粒作用后肝癌细胞出现固缩、崩解、坏死等损伤性改变。紫杉醇纳米粒组给药后1、2、3、5 d,HepG2细胞存活率分别为(76.6±5.1)%、(57.3±4.9)%、(29.7±4.4)%、(7.2±2.0)%,空白纳米粒组分别为(94.3±6.8)%、(91.0±6.6)%、(90.7±5.6)%、(87.4±5.7)%;Huh-7细胞存活率分别为(82.1±8.0)%、(63.5±5.6)%、(40.2±3.9)%、(11.4±3.6)%,空白纳米粒组分别为(93.5±7.7)%、(92.5±6.8)%、(89.0±6.2)%、(86.1±6.5)%;紫杉醇纳米粒作用的细胞存活率较相应空白纳米粒组降低(均P<0.05),且紫杉醇纳米粒作用时间越长细胞存活率降低越显著(均P<0.05)。结论紫杉醇纳米粒可在体外以可控的方式对肝癌HepG2和Huh-7细胞发挥抗肿瘤作用,为抗肝癌药物的临床应用提供了新的剂型。

Objective To prepare active and controlled paclitaxel-loaded nanoparticles, and to determine their effect on hepatocellular carcinoma HepG2 and Huh-7 cells. Methods The poly (D, L-lactide-co-glycolide) nanoparticles containing paclitaxel were prepared by o/w emulsification-solvent evaporation method. The drug encapsulation efficiency, loading efficiency and release profile rate in vitro were measured by high-performance liquid chromatography. The morphology and size of nanoparticles were observed by scanning electron microscopy and particle size analyzer, respectively. Hepatocellular carcinoma HepG2 and Huh-7 cells were co-cultured with paclitaxel nanoparticles, and the changes of cell morphology were observed by inverted microscope, and the cell viability was examined by the methyl thiazolyl tetrazolium assay. Results The nanoparticles were spherical shape with particle size 216 nm, and the drug encapsulation efficiency and loading efficiency was 87.4% and 1.79%, respectively. The drug release pattern showed an early burst release followed by a slow sustained release. Released drug from paclitaxel-loaded nanoparticles resulted in injury of HepG2 and Huh-7 cells. At 1, 2, 3 and 5 days after administration, the survival rates of HepG2 cells co-cultured with paclitaxel nanoparticles were (76.6±5.1)%,(57.3±4.9)%,(29.7±4.4)%, and (7.2±2.0)%, respectively, and the survival rates in the blank group were (94.3±6.8)%,(91.0±6.6)%,(90.7±5.6)%, and (87.4±5.7)%;the survival rates of Huh-7 cells were (82.1±8.0)%,(63.5±5.6)%,(40.2±3.9)%, and (11.4±3.6)%, respectively, and the survival rates in the blank group were (93.5±7.7)%,(92.5±6.8)%,(89.0±6.2)%, and (86.1±6.5)%. The survival rate of cells co-cultured with paclitaxel nanoparticles was lower than that of the blank group (all P < 0.05), and the cell survival rate showed a decrease change with prolonged co-cultured time (both P < 0.05). Conclusion Paclitaxel nanoparticles could play an anti-tumor effect on hepatocellular carcinoma HepG2 and Huh-7 cells in vitro in a controlled manner, providing a new dosage form for the clinical application of anti-hepatoma drugs.