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Antimicrobial Agents and Chemotherapy, September 2002, p. 2854-2864, Vol. 46, No. 9
0066-4804/02/$04.00+0     DOI: 10.1128/AAC.46.9.2854-2864.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Samarangenin B from Limonium sinense Suppresses Herpes Simplex Virus Type 1 Replication in Vero Cells by Regulation of Viral Macromolecular Synthesis

Yuh-Chi Kuo,1,2* Lie-Chwen Lin,1 Wei-Jern Tsai,1 Cheng-Jen Chou,1 Szu-Hao Kung,3 and Yen-Hui Ho1

National Research Institute of Chinese Medicine,1 Institute of Life Science, Fu-Jen University,2 Department of Medical Technology and Institute of Biotechnology in Medicine, National Yang-Ming University, Taipei, Taiwan3

Received 30 November 2001/ Returned for modification 12 March 2002/ Accepted 30 May 2002

Inhibitory effects of ethanolic extracts from 10 Chinese herbs on herpes simplex virus type 1 (HSV-1) replication were investigated. By a bioassay-guided fractionation procedure, samarangenin B (Sam B) was isolated from Limonium sinense; Sam B significantly suppressed HSV-1 multiplication in Vero cells without apparent cytotoxicity. Time-of-addition experiments suggested that the inhibitory action of Sam B on HSV-1 replication was not due to the blocking of virus adsorption. In an attempt to further localize the point in the HSV-1 replication cycle where arrest occurred, a set of key regulatory events leading to viral multiplication was examined, including viral immediate-early ({alpha}), early (ß), and late ({gamma}) gene expression and DNA replication. Results indicated that levels of glycoprotein B (gB), gC, gD, gG, and infected-cell protein 5 (ICP5) expression and gB mRNA expression in Vero cells were impeded by Sam B. Data from PCR showed that replication of HSV-1 DNA in Vero cells was arrested by Sam B. Furthermore, Sam B decreased DNA polymerase, ICP0, and ICP4 gene expression in Vero cells. Results of an electrophoretic mobility shift assay demonstrated that Sam B interrupted the formation of an {alpha}-trans-induction factor/C1/Oct-1/GARAT multiprotein complex. The mechanisms of antiviral action of Sam B seem to be mediated, at least in part, by inhibiting HSV-1 {alpha} gene expression, including expression of the ICP0 and ICP4 genes, by blocking ß transcripts such as DNA polymerase mRNA, and by arresting HSV-1 DNA synthesis and structural protein expression in Vero cells. These results show that Sam B is an antiviral agent against HSV-1 replication.

* Corresponding author. Mailing address: Rm. 912, Laboratory of Immunology, National Research Institute of Chinese Medicine, No. 155-1, Sec. 2, Li-Nung St., Shih-Pai 112, Taipei, Taiwan. Phone: (02)28201999, ext. 9111. Fax: (02)28264276. E-mail: kuo9111{at}cma23.nricm.edu.tw.

Antimicrobial Agents and Chemotherapy, September 2002, p. 2854-2864, Vol. 46, No. 9
0066-4804/02/$04.00+0     DOI: 10.1128/AAC.46.9.2854-2864.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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