Year of Publication

2013

Degree Name

Doctor of Philosophy (PhD)

Document Type

Doctoral Dissertation

College

Agriculture, Food and Environment

Department

Animal and Food Sciences

First Advisor

Dr. Melissa C. Newman

Abstract

This study investigated the antiviral, chemoprotective and proliferative effects of strawberry anthocyanins on herpes simplex virus type-1, cancerous cell lines HT-29 and AGS, and normal cell lines Hs 738.St/Int and CCD-18Co. Antiviral properties were measured by infecting vero cells from adult grivet (Cercopithecus aethiops) with herpes simplex virus type-1 (HSV-1) and treating with a concentration of 1.25-20 µg/mL of strawberry anthocyanins. Infectivity and replication were quantified for herpes simplex virus type-1 using the direct plaque assay and reporting PFU/mL. Strawberry anthocyanins (>20 µg/mL) inhibited the herpes simplex virus infectivity in vero cells by 100% (p<0.05). Strawberry anthocyanins at concentrations of 5, 10 and 20 μg/mL were reduced to 75.36, 57.98, and 31.46 percent of the control (100%) (p<0.05).

Chemoprotective and proliferative effects of strawberry anthocyanins were analyzed for the human cell lines AGS, Hs 738.St/Int, HT-29, and CCD-18Co at a concentration of 25-200 µg/mL and quantified using the sulforhodamine-B assay. Growth inhibition occurred at a level of ≥87% for treatment concentrations 100 and 200 µg/mL for the cancerous AGS and HT-29 cell lines (p<0.0001). Proliferation rates for the normal Hs 738.St/Int and CCD-18Co cell lines increased at all treatment concentrations of 25-200 μg/mL (p<0.0001); suggestingthat the observed proliferative activity may be associated with anthocyanin treatment.Strawberry anthocyanin treatment concentration worked in a dose dependent manner for the HSV-1 and the cancerous AGS and HT-29 cells. The caspase-3 assay was performed to demonstrate potential mechanism of action and confirmed thatanthocyanin treatments play a role in apoptosisby the up regulation of caspase-3.Significantdifferences were seen between the growth characteristics of cancerous cell linescompared to their equivalent normal cell lines (p<0.0001).

In summary, the antiviral findings suggest that strawberry anthocyanin extracts could be an effective topical treatment and/or prophylactic agent for oral herpetic infections (HSV-1). Also, the in vitro chemoprotective effect of strawberry anthocyanins found may be relevant to in vivo work in the future because when anthocyanins are consumed in the diet they come in direct contact with the gastrointestinal tract and may provide chemoprotection upon contact with the stomach and gastrointestinal tract’s epithelial cell layer.

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