삼백초 수용성 추출물의 Salmonella typhimurium 균에 대한 항균 및 숙주세포 내 사멸 효과 규명
Antibacterial and Intracellular Clearance Effect of Saururus chinensis Baill Water Extract against Salmonella typhimurium
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초록
살모넬라증은 대표적인 인수공통전염병의 하나로서 세포내 기생하며 질병을 유발하며 장염과 식중독 등을 유발하여 공중보건학적으로 심각한 문제를 야기하고 있다. 본 연구는 삼백초의 수용성 추출물을 (SCWE) 이용하여 숙주세포에 대한 안전성, S. typhimurium 균에 대한 항균효과 및 대식세포 내 균 증식억제 기능을 규명하였다. 본 실험을 통하여 SCWE 1, 10 및 100 μg/ml 농도로 첨가한 배지에서 RAW 264.7 체포와 24 시간 반응 후 평가해본 결과 세포독성이 인정되지 않았으며, S. typhimurium 균에 대하여 시간 경과에 따라 항균효과가 증가되는 것을 확인하였고, SCWE 처리에 의해 대식세포의 형태적 변화가 증가되는 것으로 나타났으며 (p<0.05), 살모넬라균의 탐식능력 및 대식세포 내 균 증식 억제 능력이 비 처리군에 비해 현저히 증가되는 것이 확인되었다. 또한 SCWE 처리 한 대식세포에 살모넬라균 감염을 수행하였을 때 대식세포의 nitric oxide (NO) 산생능력이 비 처리 군에 비해 저하되는 것으로 나타나, 살모넬라균에 의한 대식세포의 세포독성을 억제하는 것으로 나타났다. 종합적으로 SCWE의 숙주세포에 대한 안전성, 살모넬라균에 대한 항균효과 및 대식세포 내 균 증식 억제 효과가 있는 것으로 나타나 SCWE를 이용한 세포내 기생세균의 치료제 개발이 가능할 것으로 판단된다.
Abstract
Salmonella spp. are one of major pathogens for zoonosis in worldwide, and can replicate within host cells and generally cause enterocolitis and foodborne poisoning which represents a considerable public health burden. The present study was designated to investigate the safty for host cells, antibacterial effects of Saururus chinensis Baill water extract (SCWE) on pure culture and infection with Salmonella enterica serovar Typhimurium (S. typhimurium) in murine derived macrophage RAW 264.7 cells. The different treatment of SCWE concentration (1, 10 or 100 μg/ml) did not show any cytotoxic effect to RAW 264.7 cells for 24 h incubation. In determination of antibacterial activity of SCWE against S. typhimurium, bacterial viability was markedly decreased compared to SCWE-untreated control. In RAW 264.7 cells, SCWE significantly induced morphological change (p<0.05). In infection assay of S. typhimurium in RAW 264.7 cells pretreated with 100 μg/ml of SCWE, which are non-cytotoxic concentration, bacterial uptake ability of macrophage was increased corresponding with morphological change, whereas bacterial survival rates within macrophage was markedly reduced comparing to that of SCWE-untreated control. Furthermore, nitric oxide (NO) production in SCWE-treated cells was slightly decreased until 24 h post infection. Taken together, these findings demonstrated that SCWE have the antibacterial activity for S. typhimurium and the protective effects against S. typhimurium infection through activating murine macrophage independent on NO, suggesting that SCWE were beneficial on the disease caused by intracellularly replicating pathogens as a safe alternatives of conventional chemotherapies.
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