청도반시 추출물의 murine norovirus에 대한 항바이러스 효과
Antiviral effect of persimmon (Diospyros kaki Thunb. cv. Cheongdo-Bansi) extracts on murine norovirus
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청도반시의 항노로바이러스 효과를 검증하기 위해 메탄올, 에탄올, 아세톤, 물 추출물 및 착즙물을 제조하여 각각의 수율과 탄닌 함량을 분석한 결과, 수율은 메탄올 추출물 > 물 추출물 > 에탄올 추출물 > 아세톤 추출물 > 착즙물 순인 반면, 탄닌 함량은 착즙물 > 아세톤 추출물 > 물 추출물 > 에탄올 추출물 > 메탄올 추출물의 순으로 상반되는 결과를 보여주었다. MTT assay를 통한 세포 생존율 결과를 토대로 메탄올과 에탄올은 각각 800 μg/mL, 아세톤은 600 μg/mL, 물 추출물 및 착즙물은 200 μg/mL와 400 μg/mL로 항바이러스 효과 검증을 위한 최종 농도로 결정하였다. MNV에 대한 항노로바이러스 효과를 time-of-addition assay로 분석한 결과, 바이러스 접종 전 추출물 및 착즙물을 세포에 처리한 pre-treatment에서 물 추출물 처리구가 항노로바이러스 효과가 있음을 확인할 수 있었다. 또한, 추출물 및 착즙물과 바이러스를 동시에 접종한 co-treatment에서 아세톤, 물 추출물 및 착즙물의 처리구의 플라크 형성율이 각각 71.50±4.24%, 77.62±2.92%, 59.76±2.36%로 나타났으며, 2.14±0.20 log PFU/mL, 1.69± 0.16 log PFU/mL, 2.96±0.16 log PFU/mL로 유의적인 노로바이러스 농도 감소 효과를 보여 주었다. 그러나 바이러스 접종 이후 추출물 및 착즙물을 처리한 post-treatment에서는 5가지 추출물 및 착즙물의 노로바이러스 플라크 형성율 및 감소된 농도가 95.15-99.32%와 0.04- 0.37 log PFU/mL로 노로바이러스 농도의 감소는 보였으나, 대조구 대비 유의적인 차이를 나타내지는 않았다. 감 추출물 및 착즙물이 직접적으로 MNV에 미치는 영향을 검토한 co-treatment 결과에서는 탄닌 함량에 따라 높은 항노로바이러스 효과를 보여주었다. 결론적으로 감 착즙물의 경우, 추출 수율은 낮았으나, 탄닌 함량과 항노로바이러스 효과는 가장 높게 나타남을 확인할 수 있었다. 따라서, 향후 추가적인 연구를 통해 청도반시 착즙물의 항노로바이러스 대응 기능성 소재로서의 활용 가능성을 부각시킬 수 있을 것으로 판단된다.
Abstract
Norovirus infections are one of the most significant causes of foodborne outbreaks worldwide. This study aimed to investigate the antiviral effects of persimmon (Diospyros kaki Thunb. cv. Cheongdo-Bansi) extracts on murine norovirus (MNV). Persimmon extracts obtained by solvent extraction and squeezing, prior to freeze-drying were evaluated by determining the yield and tannin contents. The cytotoxicity and antiviral effect of persimmon extracts were determined using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and a time- of-addition assay, respectively. The yield and tannin content were in following order: methanol > water > ethanol > acetone > juice extracts and juice > acetone > water > ethanol > methanol extracts, respectively. The concentrations of methanol, ethanol, acetone, water, and juice extracts were determined to be 800, 800, 600, 200, and 400 μg/mL, respectively. The MNV titer was significantly reduced by 1.65 log plaque forming unit (PFU)/mL with 18% reduction in plaque formation in group pre-treated with water extract. Furthermore, the significant reductions in MNV titer by 2.14, 1.69, and 2.96 log PFU/mL were observed in groups co-treated with acetone, water, and juice extracts, respectively with plaque formation inhibition of 22-40%. However, there were no significant antiviral effects in the post-treated groups. This study suggests the potential use of persimmon extracts as anti-noroviral agents.
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