反义寡脱氧核苷酸在鸭体内抑制鸭乙型肝炎病毒复制与表达的研究  

Antisense oligodeoxynucleotide targeted at the pres region of DHBV could inhibit virus replication in vivo

作　　者：何丽芳[1,2,3] 吴祥甫[1,2,3] 陈常庆 陈波[1,2,3] 姚忻 袁正宏[1,2,3] 

机构地区：[1]上海医科大学医学分子病毒学卫生部重点实验室 [2]中科院上海生物化学研究所 [3]中科院上海生物工程研究中心

出　　处：《中华实验和临床病毒学杂志》1998年第1期1-4,共4页Chinese Journal of Experimental and Clinical Virology

基　　金：国家863高科技生物技术领域资助

摘　　要：目的研究反义核酸的抗病毒作用。方法设计合成了针对鸭乙型肝炎病毒（ＤＨＢＶ）前Ｓ（ＰｒｅＳ）基因区第９５１９６８位核苷酸的硫代反义寡脱氧核苷酸（ＡＳＯＤＮ），以２０μｇ／ｇ体重／日剂量对３只腹腔感染ＤＨＢＶ５２毒株后，血清ＤＨＢｓＡｇ及ＤＨＢＶＤＮＡ阳性鸭连续静脉注射１０天，同时以等体积生理盐水注射另３只感染鸭作为对照。结果对照鸭注射生理盐水后，血清ＤＨＢｓＡｇ及ＤＨＢＶＤＮＡ阳性未见明显改变，肝组织ＤＮＡＳｏｕｔｈｅｒｎ杂交在３０与２３ｋｂ左右杂交信号明显。注射ＡＳＯＤＮ鸭在１０天后，２／３鸭血清ＤＨＢｓＡｇ及ＤＨＢＶＤＮＡ量显著降低，３／３鸭肝组织中３０ｋｂ左右的杂交信号显著减弱，２３ｋｂ左右未见杂交信号。结论说明该段ＡＳＯＤＮ在鸭体内能部分抑制ＤＨＢＶ的复制与抗原表达。To study the antiviral effect of antisense oligodoxynucleotide (ASODN), ASODN phosphotioates targeted to DHBV PreS region (951968nt) (5'TATCTCCTCCATTGTTTG3') was synthesized. Two dayold ducklings were infected i.p. with 150μl of DHBV (3×108 copies/ml DHBV). After 12 dyas, 6 DHBsAg and DHBV DNA positive ducks were divided into two groups. In treated group, each duck was injected i.v. AS-ODN 20μg/gram weight/day for 10 dyas successively, while in the control group, each duck received the same volume of normal saline for 10 days. Sera from ducks prior to treatment and treated after 6, 10 days were collected, and after 10 days ducks were sacrificed and livers were examined. DHBsAg was significantly decreased in all three treated ducks after 10 days of AS-ODN treatment. One of these ducks showed decreased DHBsAg even 6 days after treatment. Two out of three treated ducks showed decreased serum DHBV DNA, while one remained unchanged. In contrast, none of the control group showed decreased DHBsAg and DHBV DNA did not change. By Southern blot, DHBV DNA in the liver significantly decreased in 2/3 of the treated ducks and decreased in the other duck. In all control ducks, DHBV DNA remained strongly positive. Data indicated that ASODN targeting at PreS region could partly inhibit the replication of DHBV.

关 键 词：乙型肝炎病毒 抗病毒治疗 复制 表达 AS-ODN 

分 类 号：R373.21[医药卫生—病原生物学]