阿德福韦酯固体脂质纳米粒体外抗病毒药效研究

Antiviral Effect of Adefovir Dipivoxil Loaded Solid Lipid Nanoparticles in Vitro

目的研究阿德福韦酯(adefovir dipivoxil,ADV)经固体脂质纳米粒(solid lipid nanoparticle,SLN)转运后的体外抗乙肝病毒药效。方法采用溶剂扩散法制备包载阿德福韦酯的固体脂质纳米粒(ADV-SLN),以紫外分光光度法测定药物包封率和载药量;以硬脂胺-异硫氰基荧光素标记SLN,通过荧光倒置显微镜观察HBVDNA转染人肝癌细胞HepG2.2.15对SLN的经时摄取情况;采用酶免疫测定试剂盒测定HbsAg,HBeAg指标,采用实时定量PCR试剂盒测定HBVDNA指标。结果ADV-SLN药物包封率为(16.72±2.81)%,载药量为(3.86±0.42)%。SLN可被HepG2.2.15有效摄取并具一定的时间依赖性。与游离药物相比,ADV经SLN转运后,对HepG2.2.15表达的HbsAg,HbeAg,HBVDNA的抑制作用均有明显增强。结论阿德福韦酯经SLN转运后,药物疗效增强,在抗乙肝病毒靶向治疗中体现出良好的应用前景。

OBJECTIVE To study antiviral effect of adefovir dipivoxil （ADV） delivered by solid lipid nanoparticles （SLN） in vitro. METHODS ADV loaded SLN （ADV-SLN） were prepared by solvent diffusion method in aqueous system, and the drug entrapment efficiency （EE） and drug loading （DL） were determined by ultraviolet spectrophotometric method at the wavelength of 260 nm. The cellular uptakes of SLN by HepG2.2.15 were investigated by ODA-FITC incorporated SLN and fluorescence microscopy. HBsAg and HBeAg were determined by commercial enzyme immunoassay kits, while HBV DNA was quantified by a commercial real-time polymerase chain reaction （PCR） kit. RESULTS EE and DL were （16.72±2.81）% and （3.86±42）% for ADV loaded SLN. HepG2.2.15 increased uptake of SLN effectively and the cellular uptake of SLN was time-dependent. Comparing with free ADV, the inhibitory effects of ADV loaded in SLN on hepatitis B surface antigen （HBsAg）, hepatitis Be antigen （HBeAg） and hepatitis B virus （HBV） DNA levels in vitro were significantly enhanced. CONCLUSION ADV delivered by NLC could significantly enhance in vitro antiviral effect, which revealed a potential application for the targeting antiviral therapy.