An in vitro system combined with an in-house quantitation assay for screening hepatitis C virus inhibitors.
Hepatitis C virus (HCV) infection is a serious global health problem. Interferon-alpha (IFN-alpha) and ribavirin have demonstrated efficacy in the treatment of HCV infection; however, these therapies display many side effects. To screen the anti-HCV compounds from plants, we established an in vitro model for inoculation of HCV by centrifugation-facilitated method. The HCV RNA molecules were then quantitated by nested competitive reverse transcription-polymerase chain reaction (cRT-PCR) using fluorescein-labeled primers, and analyzed by ABI Prism 310. The positive and negative strands of HCV RNA were detectable in Vero cells on Day 7 post-infection, suggesting that the HCV RNA was present in the cell model system. The cell culture system was further used to screen the anti-HCV activities of 4 Chinese herbal formulas and 15 formula components. IFN-alpha showed an antiviral effect. The formulas exhibited no anti-HCV activities, while Arnebia euchroma, Thlaspi arvense, and Poncirus trifoliata displayed anti-HCV activities. Therefore, these results pointed out the possibility by using the cell culture system established in this study to screen the herb extracts for their anti-HCV activities.
Ho
Tin-Yun
TY
Institute of Chinese Medical Science, China Medical College, ROC, Taichung, Taiwan.
Wu
Shih-Lu
SL
Lai
I-Lu
IL
Cheng
Ken-Sheng
KS
Kao
Shung-Te
ST
Hsiang
Chien-Yun
CY
D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology*; D002498:Centrifugation; D002522:Chlorocebus aethiops; D016174:Hepacivirus / Q000187:drug effects*; D006526:Hepatitis C / Q000821:virology; D006801:Humans; D008826:Microbial Sensitivity Tests / Q000379:methods; D010936:Plant Extracts / Q000494:pharmacology*; D010946:Plants, Medicinal / Q000737:chemistry*; D012367:RNA, Viral / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D014709:Vero Cells / Q000821:virology
D000998:Antiviral Agents; D010936:Plant Extracts; D012367:RNA, Viral