Abstract
In this study, the biological activity of water and ethanol extracts from Chrysanthemum incidicum Linne by ultrafine grinding for functional food source are examined. The content of phenolic compounds from Chrysanthemum incidicum Linne were the highest when extracted for 6 hr with 70% ethanol. The extraction yield of water and ethanol extracts were $7.12{\pm}1.61$ mg/g and $7.51{\pm}2.14$ mg/g, respectively. With ultrafine grinding, water and ethanol extracts were $8.63{\pm}1.15$ mg/g and $9.33{\pm}1.35$ mg/g, respectively. In determining anti-oxidative activity of Chrysanthemum incidicum Linne extracts, DPPH of normal grinding extracts was 83.52% and ultrafine grinding was 92.37%. In ABTS radical cation decolorization, normal grinding, fine grinding, and ultrafine grinding extracts were 90% or higher. In antioxidant protection factor (PF), water and ethanol extracts of ultrafine grinding showed relatively high anti-oxidative activities of each 1.82 PF and 2.16 PF, respectively. The TBARS value of ultrafine grinding extracts were lower than normal grinding and fine grinding extracts. The inhibition activity on xanthin oxidase of Chrysanthemum incidicum Linne extracts was 67.53% in ultrafine grinded water extracts and 83.45% in ultrafine grinded ethanol extracts. Inhibition on xanthin oxidase of ethanol extracts showed a higher inhibition effect than water extracts, and ultrafine grinding was higher than normal grinding. In angiotensin converting enzyme inhibition activity, ultrafine grinding water extract was 24% or higher, and ethanol extract was 34% or higher. The elastase inhibition activity of ultrafine grinding extract was 25.56%, which was higher than 20.34% of fine grinding extracts. Water extracts did not show hyaluronidase inhibition activity but ethanol extracts showed 35% of hyaluronidase inhibition activity. The determining expression inhibition of iNOS and COX-2 protein in macrophage by Chrysanthemum incidicum Linne extracts with a Western blot analysis, iNOS and COX-2 protein expression inhibition by Chrysanthemum incidicum Linne ethanol extracts were 40% and 15%, respectively at 100 ${\mu}g/mL$ concentration. The inhibitory patterns of iNOS and COX-2 protein expression was concentration dependent. The result suggests that Chrysanthemum incidicum Linne extracts by ultrafine grinding may be more useful than normal grinding as potential sources due to anti-oxidation, angiotensin converting enzyme and xanthine oxidase inhibition, anti-inflammation effect.
감국 phenolic compound의 추출을 위한 최적 추출조건은 70% ethanol로 6시간 추출이 최적이었다. 그때 추출수율은 물 추출물이 $7.12{\pm}1.61$ mg/g, ethanol 추출물이 $7.51{\pm}2.14$ mg/g이었고 초미세분쇄 했을 경우 물 추출물이 $8.63{\pm}1.15$ mg/g, ethanol 추출물이 $9.33{\pm}1.35$ mg/g이었다. 감국 추출물의 항산화력 측정에서 DPPH는 일반분쇄 추출물이 83.52%, 초미세분쇄 추출물이 92.37%이었다. ABTS radical cation decolorization의 경우 일반분쇄, 미세분쇄 및 초미세분쇄 추출물 모두 90% 이상이었다. Antixodant protection factor(PF)의 경우 초미세분쇄 한 감국의 물과 에탄올 추출물에서 1.82 PF와 2.16 PF의 높은 항산화력을 나타내었다. TBARS도 일반분쇄 및 미세분쇄에 비해 초미세분쇄 한 감국 추출물에서 더 낮은 TBARS값을 나타내었다. 감국 추출물의 xanthin oxidase 저해효과는 초미세분쇄 물 추출물이 67.53%, ethanol 추출물이 83.45%였다. Xanthin oxidase 저해는 물 추출물보다는 ethanol 추출물이, 초미세분쇄 추출물이 일반 추출물에 비해 효소억제효과가 더 우수한 결과를 나타내었다. Angiotensin converting enzyme 저해효과는 초미세분쇄 추출물의 경우 물 추출물에서 24% 이상의 억제효과를 나타내었고, ethanol 추출물은 34%의 억제효과를 나타내었다. 감국 추출물의 elastase 억제효과는 미세분쇄 추출물의 저해율 20.34%에 비하여 초미세분쇄 추출물의 억제율이 25.56%로 더 우수한 기능성을 나타내었다. Hyaluronidase의 경우 감국 물 추출물에서는 억제효과가 관찰되지 않았으며, ethanol 추출물에서만 35%정도의 염증억제효과를 보여주었다. 감국 추출물에 의해 대식세포에서 iNOS와 COX-2 단백질 발현억제 효과를 Western blot analysis로 측정한 결과 감국 에탄올 추출물에 의해 iNOS와 COX-2 단백질 발현이 100 ${\mu}g/mL$의 농도에서 각각 40%와 15%의 발현억제를 나타내었고, 농도 의존적으로 억제됨을 확인하였다. 본 연구결과 초미세분쇄에 의한 감국 추출물은 일반분쇄 추출물에 비해 고혈압과 통풍억제 효과가 확연히 높아짐을 알 수 있었다.