Ethnomedicinal Plant Stephania hernandifolia and its Active Fraction 2-
Chloroethyl Linoleate Inhibits HSV-2 Infection by Blocking Viral Immediate
Early and Early Transcription

Joy      Mondal; Chiranjit      Patra; Ananya   Das   Mahapatra; Keshab   Chandra   Mandal; Debprasad      Chattopadhyay

doi:10.2174/1574885517666220512165130

Abstract

Aims: The aim of this study is validation of antiviral activity of Stephania hernandifolia against HSV-2.

Background: Ethnomedicinal plant Stephania hernandifolia, traditionally used for the management of skin, digestive and nerve ailments demonstrated significant anti-HSV-1 activity; similar to Stephania cepharantha having neuroinflammatory and anti-HSV activities.

Objectives: Thus, the present study aimed to validate the potential of the most active fraction-2 (F-2) of S. hernandifolia against HSV-2 in vitro, along with the underlying mode or mechanism of action.

Methods: The standardized F-2 was characterized by GC-MS, ¹H-NMR, Mass and FTIR spectroscopy. Cytotoxicity (CC₅₀) and antiviral activity (EC₅₀) were evaluated by MTT and Plaque reduction assay. To determine the mode of action, we have used time-of-addition, virus inactivation, and entry (attachment and penetration) assays, followed by semiquantitative PCR. Furthermore, the protein expression levels of immediate early (IE) and early (E) gene products of drug-treated virions were measured by Western blot.

Results: The results showed that HSV-2G and ICMR/VU-2012/20, the clinical isolate of HSV-2, were inhibited by F-2 at EC₅₀ of 20.0 and 20.43 μg/ml respectively, with Selectivity Index (SI) of 12. Timeof- addition assay showed that F-2 significantly inhibited HSV-2 infection in Vero cells at 4-8 h posttreatment. The infectivity of the virion was lost within 1h of exposure to F-2 (EC₅₀ and EC₉₉). Furthermore, semi-Q-PCR and Western blot studies demonstrated significant downregulation of IE and E gene products. The characterization revealed that 2-chloroethyl linoleate is the lead compound in the F-2 fraction.

Conclusion: Thus, our results showed that the bioactive fraction F-2 inhibits both IE and E transcription of HSV-2.

Keywords: Ethnomedicine, Stephania hernandifolia, herpes simplex virus type 2, antiviral activity, GC-MS, mass spectra, NMR, FTIR.

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DOI https://dx.doi.org/10.2174/1574885517666220512165130	Print ISSN 1574-8855
Publisher Name Bentham Science Publisher	Online ISSN 2212-3903

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Ethnomedicinal Plant Stephania hernandifolia and its Active Fraction 2- Chloroethyl Linoleate Inhibits HSV-2 Infection by Blocking Viral Immediate Early and Early Transcription

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