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目的制备载siRNA阳离子脂质体并对其进行制剂学评价。方法利用siRNA与溴化乙锭结合能产生荧光的原理,采用预染溴化乙锭(Ethidium Bromide,EB)琼脂糖凝胶,氨基丁三醇-硼酸-乙二胺四乙酸(Tris-Boric acid-EDTA,TBE)电泳缓冲液将游离siRNA分离并通过凝胶成像分析系统转换数据进行定性和定量,计算载siRNA阳离子脂质体包封率,并考察其阴离子的抵御能力和血清稳定性;利用Malver粒径和zeta电位测定仪对不同载siRNA阳离子脂质体进行评价。结果琼脂糖凝胶电泳法的线性为0.665~13.30 mg·L-1(R2=0.988 3),回收率为97%~103%,精密度RSD<2%,阳离子脂质体对siRNA包封率较高,且具有良好的抵御阴离子能力和较强的血清稳定性;载siRNA阳离子脂质体的粒径和zeta电位变化与文献报道一致。结论所制备的载siRNA阳离子脂质体体外性质均良好。
Objective To prepare siRNA cationic liposomes and evaluate their formulation. Methods The principle of using siRNA combined with ethidium bromide to produce fluorescence was studied by using ethidium bromide (EB) agarose gel, triethanolamine-boric acid-ethylenediaminetetraacetic acid (Tris-Boric acid -EDTA, TBE) electrophoresis buffer was used to separate the free siRNA and qualitatively and quantitatively through the gel imaging analysis system conversion data to calculate the entrapment efficiency of siRNA cationic liposomes and investigate the anion’s resistance and serum stability; The Malver particle size and zeta potential meter were used to evaluate the different siRNA-loaded cationic liposomes. Results The linearity of agarose gel electrophoresis was 0.665-13.30 mg · L-1 (R2 = 0.9883), the recoveries were 97% -103%, RSD <2% High, and has good ability to resist the anion and strong serum stability; contained siRNA cationic liposomes and zeta potential changes consistent with the literature. Conclusion The prepared siRNA cationic liposomes have good in vitro properties.