阳离子脂质体对反义寡核苷酸抗乙型肝炎病毒作用的促进

目的 :考察阳离子脂质材料 3β N (N′,N′ 二甲基胺乙基 )胺基甲酰基 ]胆固醇 (3β N (N′,N′ dimethylaminoethane) carbamoyl]cholesterol,DC chol)与反义寡核苷酸 (antisenseoligodeoxynucleotide ,asODN)的结合能力及asODN阳离子脂质体的细胞毒性和抗病毒活性。方法 :以DC chol从水相中萃取asODN的萃取率评价不同条件下DC chol与asODN的作用力 ;通过四唑盐比色法检测含DC chol的阳离子脂质体对HepG2 2 .2 .1 5的细胞毒性 ;以酶联免疫法 (ELISA)检测细胞上清培养液中HBsAg和HBeAg的含量 ,考察不同asODN阳离子脂质体的抗病毒活性。结果 :当正负电荷比 >0 .6时 ,DC Chol能有效的从水相中萃取asODN (萃取率 80 %～ 1 0 0 % ) ;碱性条件和强离子 (Na+ /Cl-)的存在不利于DC Chol与asODN的作用 ;DC chol阳离子脂质体具有一定的细胞毒性 ,在低质量浓度时 (0 .84～ 2 6 .94mg·L-1 )脂质体的细胞毒性与DC chol浓度成正比 ;当asODN在 1 .2 5～ 5 .0 0 μmol·L-1时 ,其抗病毒活性与剂量相关 ;脂质体可提高asODN对乙肝病毒的抗病毒活性 ,其中阳离子脂质体对asODN抗病毒活性的促进作用较明显 (从 6 0 %到 95 % )。结论 :表面修饰DC Chol阳离子脂质体能有效地促进asODN的抗乙肝病毒活性。

Cationic liposomes enhance the inhibitory effects of antisense oligodeoxynucleotide on hepatitis B virus

OBJECTIVE: To study the interaction between 3beta-N-(N', N'-dimethylaminoethane)-carbamoyl] cholesterol (DC-chol), a cationic lipid, and antisense oligodeoxynucleotide (asODN), and the cytotoxicity to HepG(2) 2.2.15 cells and the inhibitory effects of asODN cationic liposomes on hepatitis B virus. METHODS: Extract efficiency was used to estimate the interaction between DC-chol and asODN under different conditions; MTT was applied to examine the infection of DC-chol concentration on the cytotoxicity of treated cells; By ELISA technology, we examined the contents of HBsAg, and HBeAg in the culture medium, and evaluated the accelerate efficiency of neutral liposome and cationic liposomes to the inhibitory effects of asODN on hepatitis B virus, and the infection of charge ratio on the inhibitory effects of asODN cationic liposomes. RESULTS: When the charge ratio >0.6, DC-chol could efficiently extract the asODN from the aqueous phase; The alkaline condition and the presence of Na(+), Cl(-) ions prevented the efficient interaction between DC-chol and the asODN; DC-chol cationic liposomes had some toxicity to HepG(2) 2.2.15 cells, at low concentration (0.84-26.94 mg x L(-1)), and the toxicity of DC/chol was linear with the concentration of DC/chol; Within 1.25-5.00 micromol x L(-1), the inhibition efficacy of asODN was correlated to its concentration; liposomes could enhance the inhibition efficacy of asODN on hepatitis B virus, and cationic liposomes could enhance much more (from 60% to 95%). CONCLUSION: Surface-modified DC-chol cationic liposomes can efficiently enhance the anti-hepatitis B virus efficiency of asODN in vitro.