Date of Award

Spring 5-16-2015

Degree Type

Thesis

Degree Name

MS Microbiology

Department

Biology

Advisor

Tin-Chun Chu, Ph.D

Committee Member

Jane L. Ko, Ph.D

Committee Member

Daniel B. Nichols, Ph.D

Committee Member

Allan D. Blake, Ph.D

Keywords

HSV-1 anti-viral chinese knotweed

Abstract

HSV-1 is an opportunistic virus responsible for infecting the majority of the global population. The ability of this virus to strategically alternate between lytic and latent states has made the development of effective treatments challenging. Polygonum multiflorum (Chinese Knotweed, CK) is a perennial plant local to the south central regions of China. The thunb (root) constituents of this compound have been used in traditional Chinese medicines to treat a broad range of illnesses for centuries. In the present study, CK root extracts were evaluated for potential anti-HSV-1 activity. Cytopathic effect (CPE) monitoring studies indicate that Vero cells infected with HSV-1 pre-treated with 0.1% CK showed no signs of CPE after seven days. The potential cytotoxic effect of the compound was also assessed via cytotoxicity and cell proliferation assays. These results indicate that a 1% CK concentration displayed no signs of cytotoxicity. In order to quantify viral inhibition and determine the compounds potential mechanism of action (MOA), plaque reduction and binding assays were performed. These assays suggest that 0.1% CK concentrations are able to reduce viral infectivity by >99% by inhibiting viral binding. Similarly, viral tracking assays using flow cytometry and fluorescent microscopy suggest that as the CK concentrations are increased, HSV-1 infection is decreased. Furthermore, molecular analysis from real-time quantitative PCR (qPCR) assays suggest that 0.1% CKs concentrations are able to inhibit >98% of HSV-1 viral entry when compared to untreated HSV-1 positive controls. Overall, CKs ability to prevent in vitro HSV-1 infection may provide clinicians a potential natural alternative and/or synergistic agent to current HSV-1 therapies.

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