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目的观察硫代反义寡核苷酸 (S- ASODN)体外对 HDV的抑制作用。方法在 HDV/ HBV感染人胎肝细胞中加入不同浓度的针对 HDV Stem 区 6 84～ 6 98位核苷酸的 15聚 S- ASODN,分别采用 EL ISA和斑点杂交法检测上清液中 HDAg和细胞中 HDV RNA。结果 HBs Ag、HDAg在感染后第 2 d至第 16 d均可测出 ,以第 4d至第 12 d达高峰。加入 S- ASODN(2、4、6 μmol/ L)后 2 d,肝细胞中 HDV复制受到抑制 ,培养上清中 HDAg分泌量也减少 ,其抑制率呈剂量依赖关系 ;在同等剂量时(4μmol/ L) ,S- ASODN作用 2 d、4d、6 d,对 HDV抑制率大至相同。结论 HDV在原代培养人胎肝细胞中能稳定复制和表达至少达 12 d;S- ASODN能有效抑制 HDV/ HBV感染人胎肝细胞中 HDV复制与表达。本研究为进一步在动物体内研究 S- ASODN抗HDV作用和向临床过渡提供了有益的实验依据。
Objective To observe the inhibitory effect of S-antisense oligonucleotide (S-ASODN) on HDV in vitro. Methods Different concentrations of 15-poly (S-ASODN) against HDV / HBV infected human fetal hepatocytes were isolated from 6,848 to 698 nucleotides of HDV Stem region. The levels of HDAg and HDV RNA in cells. Results HBsAg and HDAg were detected at the 2nd day to the 16th day after infection, reaching the peak on the 4th day to the 12th day. The HDV replication in hepatocytes was inhibited 2 d after addition of S-ASODN (2,4,6 μmol / L), and the HDAg secretion in the culture supernatant was also reduced. The inhibition rate was in a dose-dependent manner. At the same dose (4 μmol / L), S-ASODN effect 2d, 4d, 6d, HDV inhibition rate as high as the same. Conclusion HDV can stably replicate and express in primary cultured human fetal hepatocytes for at least 12 days. S-ASODN can effectively inhibit the replication and expression of HDV in HDV / HBV infected human fetal hepatocytes. This study provides a useful experimental evidence for the further study of the anti-HDV effect of S-ASODN in animals and its clinical transition.