Beijing Institute of Transfusion Medicine, Beijing, China.
PLoS One. 2010 Nov 18;5(11):e14043. doi: 10.1371/journal.pone.0014043.
The development of small molecule inhibitors of hepatitis C virus (HCV) core protein as antiviral agents has been intensively pursued as a viable strategy to eradicate HCV infection. However, lack of a robust and convenient small animal model has hampered the assessment of in vivo efficacy of any antiviral compound.
METHODOLOGY/PRINCIPAL FINDINGS: The objective of this work was to develop a novel method to screen anti-core protein siRNA in the mouse liver by bioluminescence imaging. The inhibitory effect of two shRNAs targeting the highly conserved core region of the HCV genome, shRNA452 and shRNA523, was examined using this method. In the transient mouse model, the effect of shRNA-523 was detectable at as early as 24 h and became even more pronounced at later time points. The effect of shRNA-452 was not detectable until 48 h post-transduction. In a stable mouse model, shRNA523 reduced luciferase levels by up to 76.4±26.0% and 91.8±8.0% at 6 h and 12 h after injection respectively, and the inhibitory effect persisted for 1 day after a single injection while shRNA-Scramble did not seem to have an effect on the luciferase activity in vivo.
CONCLUSIONS/SIGNIFICANCE: Thus, we developed a simple and quantitative assay for real-time monitoring of HCV core protein inhibitors in mice.
开发丙型肝炎病毒（HCV）核心蛋白小分子抑制剂作为抗病毒药物已成为根除 HCV 感染的可行策略。然而，缺乏强大而方便的小动物模型阻碍了对任何抗病毒化合物体内疗效的评估。
方法/主要发现：本工作的目的是开发一种通过生物发光成像筛选小鼠肝脏中抗核心蛋白 siRNA 的新方法。使用该方法检测了针对 HCV 基因组高度保守核心区的两种 shRNA（shRNA452 和 shRNA523）的抑制作用。在瞬时小鼠模型中，shRNA-523 的作用在转染后 24 小时即可检测到，并且在稍后的时间点更为明显。shRNA-452 的作用直到转染后 48 小时才检测到。在稳定的小鼠模型中，shRNA523 在注射后 6 小时和 12 小时分别将荧光素酶水平降低了 76.4±26.0%和 91.8±8.0%，单次注射后抑制作用持续 1 天，而 shRNA-Scramble 似乎对体内荧光素酶活性没有影响。
结论/意义：因此，我们开发了一种简单、定量的检测方法，用于实时监测小鼠体内 HCV 核心蛋白抑制剂。
