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目的：在细胞水平上观察核酶对ＨＢＶ基因表达的作用。方法：将在体外具有切割活性的核酶ＲＣＰ的基因构建在表达载体ｐＳＶＬ中，获得重组质粒ＲＣＰ／ｐＳＶＬ，运用脂质体介导的基因转染技术将重组核酶基因引入ＨｅｐＧ２２１５细胞，并以空载体ｐＳＶＬ转染组为对照，通过固相放射免疫测定方法检测细胞培养上清的ＨＢｓＡｇ及ＨＢｅＡｇ分泌量，以此观察核酶ＲＣＰ对ＨＢＶ基因表达的影响。结果：在暂时性表达系统中，针对Ｐ基因５＇－端２３６０位点的核酶ＲＣＰ对ＨｅｐＧ２２１５细胞中ＨＢｓＡｇ和ＨＢｅＡｇ分泌量的抑制率分别是２６．７％、２４．８％。结论：锤头状核酶ＲＣＰ在细胞水平上能一定程度抑制ＨＢＶ基因的表达。
Objective: To observe the effects of ribozymes on HBV gene expression at the cellular level. METHODS: The RCP gene with cleavage activity in vitro was constructed in expression vector pSVL to obtain recombinant plasmid RCP / pSVL. The recombinant ribozyme gene was introduced into HepG2215 cells by liposome-mediated gene transfection technique. Empty vector pSVL transfection group as a control, by solid-phase radioimmunoassay detection of cell culture supernatant of HBsAg and HBeAg secretion, in order to observe the ribozyme RCP on HBV gene expression. Results: In the transient expression system, the inhibitory rates of RCP against HepG2215 cells at the 2360 site of P gene were 26.7% and 24.8%, respectively. Conclusion: The hammerhead ribozyme RCP can inhibit the expression of HBV gene to some extent at the cellular level.