硫代反义寡脱氧核苷酸在树体内对HDV复制与表达的抑制作用

Inhibitory effects of antisense phosphorothioate oligodeoxynucleotide on replication and expression of HDV in Tupaiae

目的：在树（Ｔｕｐａｉａ）体内观察硫代反义寡脱氧核苷酸（Ｓ－ＡＳＯＤＮ）对ＨＤＶ复制、表达的抑制作用。方法：树同时接种ＨＢＶ阳性血清０．１ｍｌ、ＨＤＶ阳性血清０．３ｍｌ后，将１６只ＨＤＶ／ＨＢＶ感染成功的树随机分为两组，给药组８只树按每只每次经尾静脉注射Ｓ－ＡＳＯＤＮ３ｍｇ，隔日１次，共７次。对照组中２只注射等量生理盐水，另６只不作处理。注射后５、１０、１５、２５ｄ取血及肝组织采用免疫组化、斑点杂交及原位杂交法检测ＨＤＶＡｇ及ＨＤＶＲＮＡ。结果：于注射结束时（１５ｄ）给药组有７只树肝内ＨＤＶＡｇ及ＨＤＶＲ－ＮＡ转为阴性，对照组８只树中仅１只转为阴性。停止使用Ｓ－ＡＳＯＤＮ１０ｄ后，给药组２只阴转树肝内又可检出ＨＤＶＡｇ和ＨＤＶＲＮＡ，对照组仍有７只可检出。结论：Ｓ－ＡＳＯＤＮ在树体内能有效抑制ＨＤＶ复制及表达，但作用并不完全，可能与用药剂量不足。

Objective: To investigate the inhibitory effects of antisense phosphorothioate oligodeoxynucleotide (S ASODN) on replication and expression of HDV in Tupaiae. Methods: Tupaiae were co inoculated with 0.1 ml HBV positive serum and 0.3 ml HDV positive serum. Sixteen Tupaiae infected with HDV/HBV were then randomized into two groups with equal number for each: the treated group (TG) and the control group (CG). The animals in TG were treated with injection of 3 mg S ASODN via the caudal vein once every two days for 7 times. In CG, 2 animals were treated with normal saline in the same way and the left 6 were not given any treatment. Blood samples and liver specimens were collected on the 5th, 10th, 15th and 25th day after treatment. HDVAg and HDV RNA in the liver were detected with immunohistochemistry and in situ hybridization. Results: On the 15th day after treatment, HDVAg and HDV RNA in the liver became negative in 7 out of 8 in TG while only 1 out of 8 in CG. On the 25th day after treatment, they were positive in 2 out of the 7 negative ones in TG and in 7 out of 8 in CG. Conclusion: S ASODN can effectively inhibit the replication and expression of HDV in Tupaiae. However, the inhibitory effects are not strong enough. The reason might be related to the inadequate dosage or treatment duration and indirect modification to the target.