In vitro activity of cepharanthine hydrochloride against clinical wild-type and lamivudine-resistant hepatitis B virus isolates.
In vitro activity of cepharanthine hydrochloride against clinical wild-type and lamivudine-resistant hepatitis B virus isolates.
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盐酸千金藤碱对临床野生型和拉米夫定耐药乙型肝炎病毒分离株的体外活性
DOI：
10.1016/j.ejphar.2012.02.030
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发表时间：
2012-05-15
影响因子：
5
通讯作者：
Wang QD
中科院分区：
文献类型：
--
作者：
Zhou YB;Wang YF;Zhang Y;Zheng LY;Yang XA;Wang N;Jiang JH;Ma F;Yin DT;Sun CY;Wang QD
乙型肝炎病毒（HBV）感染在世界范围内造成重大公共卫生问题。目前治疗HBV的抗病毒药物（如拉米夫定）的临床局限性是在长期抗病毒治疗期间出现耐药病毒株。盐酸头孢酞菁（CH）是一种天然生物碱衍生化合物，据报道对多种病毒具有强效活性。本研究旨在评估CH在瞬时转染细胞中对临床野生型和拉米夫定耐药HBV分离株的体外活性。从拉米夫定治疗前和病毒突破时收集的血清样本中提取HBV DNA，并通过聚合酶链反应（PCR）扩增。这些扩增子被克隆到一个新的表达载体pHY106中，该载体在细胞转染后可以启动细胞内HBV复制周期。将克隆的扩增子转染HepG2细胞后，进行药敏试验。采用酶联免疫吸附试验（ELISA）检测病毒抗原HBeAg水平。采用实时荧光定量PCR （Quantitative real-time PCR, Q-PCR）检测细胞内HBV DNA含量。通过逆转录PCR （RT-PCR）分析了HBV复制所需的宿主蛋白热应激同源蛋白70 (Hsc70)，探讨了CH可能的抗病毒机制。结果表明，CH对野生型和拉米夫定耐药HBV临床分离株的复制和HBeAg产生均呈剂量依赖性。Hsc70 mRNA也显著下调。综上所述，CH对野生型和拉米夫定耐药HBV临床分离株均有活性，其活性可能与其抑制宿主Hsc70有关。
Hepatitis B virus (HBV) infection causes major public health problems worldwide. The clinical limitation of current antiviral drugs for HBV, such as lamivudine, is the emergence of drug-resistant viral strains during prolonged antiviral therapy. Cepharanthine hydrochloride (CH), a natural alkaloid-derived compound, has been reported to possess potent activity against various viruses. The present study was performed to evaluate the in vitro activity of CH against clinical wild-type and lamivudine-resistant HBV isolates in transiently transfected cells. HBV DNA was extracted from serum samples collected both before lamivudine therapy and at the time of viral breakthrough and was amplified by polymerase chain reaction (PCR). The amplicons were cloned into a novel expression vector, pHY106, which can initiate the intracellular HBV replication cycle after cell transfection. Following transfection of the cloned amplicon into HepG2 cells, a drug susceptibility assay was performed. The level of viral antigen, HBeAg, was determined by enzyme-linked immunosorbent assay (ELISA). Quantitative real-time PCR (Q-PCR) was used for determining the amount of intracellular HBV DNA. Heat stress cognate 70 (Hsc70), a host protein required for HBV replication, was also analyzed by reverse transcription PCR (RT-PCR) to explore the possible antiviral mechanism of CH. The results showed that CH inhibited replication and HBeAg production by either wild-type or lamivudine-resistant HBV clinical isolates in a dose-dependent manner. The Hsc70 mRNA was also downregulated significantly. In conclusion, CH is active against both wild-type and lamivudine-resistant HBV clinical isolates, and its activity may be associated with its inhibition of host Hsc70.
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影响因子：
13.5
作者：
Hongthanakorn, Chanunta;Chotiyaputta, Watcharasak;Lok, Anna S. F.
通讯作者：
Lok, Anna S. F.
影响因子：
--
作者：
Baba, M;Okamoto, M;Ono, M
通讯作者：
Ono, M
影响因子：
4.9
作者：
Brunelle, M. N.;Lucifora, J.;Zoulim, F.
通讯作者：
Zoulim, F.
影响因子：
12.7
作者：
Suzuki, F;Akuta, N;Kumada, H
通讯作者：
Kumada, H
影响因子：
2.5
作者：
Westland, CE;Yang, H;Xiong, S
通讯作者：
Xiong, S