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율무근의 식물화학적 성분 연구 및 Tyrosinase 저해 활성
최윤혁 ( Yun-hyeok Choi ),최춘환 ( Chun Whan Choi ),이재연 ( Jae Yeon Lee ),안은경 ( Eun-kyung Ahn ),오좌섭 ( Joa Sub Oh ),홍성수 ( Seong Su Hong ) 한국응용생명화학회 2017 Journal of Applied Biological Chemistry (J. Appl. Vol.60 No.1
본 연구는 천연 미백소재 탐색을 위하여 율무(C. lachryma-jobivar. ma-yuen) 부산물 추출물의 tyrosinase 저해활성을 검증하였으며, 그 중에서 뿌리 추출물은 159.3 μg/mL의 IC50 값을 나타내었다. 율무 뿌리 추출물의 순차적 용매 분획에 대한 활성 검증 후 가장 활성이 우수한 EtOAc 분획물에 대하여 DiaionHP-20 column chromatography, MPLC및 preparative HPLC를 수행해 율무근으로부터 여섯 개의 화합물을 정제하였고, 이 물질들의 구조 분석은 LC-MS와 NMR 데이터 해석을 바탕으로 진행하여 각각 (+)-icariol A<sub>2</sub> (1), zhepiresionol (2), 4-hydroxybenzaldehyde(3), trans-ρ-coumaric acid (4), N-(2-hydroxy-4-methoxyphenyl)-2-hydroxyacetamide (5), coixol (6)로 구조를 규명하였다. 화합물 1은 이 식물에서 처음으로 분리되었으며, 화합물 5는 자연계로부터 처음으로 분리된 화합물로 확인하였다. 이들 화합물 중 4-hydroxybenzaldehyde (3), trans-ρ-coumaric acid (4)와 coixol (6)은 순서대로 707.4, 6.5, 62.4 μM의 IC<sub>50</sub>값을 나타내어 대조군으로 사용한 arbutin (IC<sub>50</sub> =618.7 μM)과 비교시 유사하거나 우수한 활성을 확인하였다. 이로 미루어 보아, 율무근 추출물과 활성물질의 tyrosinase 저해활성을 확인할 수 있었으며, 율무근 추출물의 미백소재로서의 적용 가능성을 확인할 수 있었다. In the course of screening tyrosinase inhibitory activity, EtOAc-soluble fraction of Coix lachryma-jobi var. mayuen Stapf. (Gramineae) roots showed significant inhibition. Further fractionation of the EtOAc-soluble fraction resulted in six compounds, which were identified as (+)-icariol A<sub>2</sub> (1), zhepiresionol (2), 4-hydroxybenzaldehyde (3), trans-ρ-coumaric acid (4), N-(2- hydroxy-4-methoxyphenyl)-2-hydroxyacetamide (5), and coixol (6). The chemical structures of these compounds were identified on the basis of spectroscopic methods (MS, 1D and 2D NMR) and comparison with literature values. Compound 1 was first reported from this plant. Also, this is the first time that the isolation of compound 5 has been reported from nature source. Among the isolated compounds, compounds 4 and 6 showed enzyme inhibitory activity, with IC<sub>50</sub> values of 6.5 and 62.4 μM, respectively, in comparison with these of positive control, arbutin.
호두 추출물의 항산화 활성과 신피질에서 세포 손상과 지질과산화 방지효과
배계선,황을철,권채화,김순희,최춘환,Bae Kae Sun,Hwang Eul Chul,Kwon Chae Hwa,Kim Soon Hee,Choi Chun Whan 한국생명과학회 2005 생명과학회지 Vol.15 No.1
본 연구에서는 호두박 분획층의 항산화 활성을 검색하기 위해 MeOH, $CH_2Cl_2$, EtOAc, BuOH 그리고 $H_2O$ 분획으로 추출하여 수행하였고, 각 분획층의 산화적 세포손상과 지질 과산화의 방지 효과를 in vitro에서 확인하였다. 그 결과 호두박의 free radical (DPP보 radical) 소거 활성은 EtOAc층에서 가장높게 나타났고, 활성산소종 억제율과 peroxynitrite $(ONOO^-)$ 소거 활성은 $CH_2Cl_2$층을 제외한 모든 분획층에서 높게 나타났다. 신피질 절편에 t-BHP의 처리 시 LDH의 방출과 지질과 산화를 증가시켰고 이러한 변화는 호두의 MeOH, EtOAc, n-BuOH 분획에 의해서 완전하게 방지되었다. 이러한 결과는 호두 추출물이 t-BHP에 의한 신장 세포 손상에 효과적이고 이러한 효과는 항산화력에 의한 것으로 추측된다. 또한 이러한 결과는 호두 추출물에서 많은 질병의 원인이 되고 있는 산화적 스트레스를 방지할 수 있는 효과적인 약물 개발의 가능성을 시사한다 To investigate the antioxidant activity of extract from the raw walnut, Juglans sinensis Dode, we prepared five fractions (methanol (MeOH), dichloromethane $(CH_2Cl_2)$, ethyl acetate (EtOAc), n-buthanol (n-BuOH) and dehydrogen monooxide $(H_2O)$ fractions) and examined. The effect of walnut extract on the oxidative stress was investigated in vitro. The DPPH (2,2-Di (4-tert-octylphenyl)-1-picrylhydrazyl) free radical scavenging activity of extract from raw walnut was shown in the following order: $EtOAc\;fraction<n-BuOH fraction< MeOH fraction<CH_2Cl_2\;fraction<H_2O$ layer. The result showed that the highest activity $(0.56{\mu}g/ml,\;IC_{50}.)$ was observed in EtOAc fraction, whereas n-BuOH fraction, MeOH fraction, $CH_2O_2$ fraction and $H_2O$ layer of $IC_{50}$ were $2.34{\mu}g//ml,\;3.88{\mu}g/ml,\;8.06{\mu}g/ml,\;and\;8.19{\mu}g/ml$, respectively. The radical scavenging activity assay of each fraction showed that the antioxidative activity was observed in the following order: EtOAc fraction $(74.27\pm1.56\%)>MeOH\;fraction\;(60.76\pm3.4\%)>n-BuOH\;fraction\;(59.32\pm0.88\%)>H_2O\;layer\;(41.69\pm2.06\%)$. These results revealed that all fractions, except for $CH_2Cl_2$ fraction, showed high antioxidative activity. Furthermore, the peroxynitrite $(ONOO^-)$ scavenging activity was assayed in each fraction. The result showed that the $ONOO^-$ scavenging activity of EtOAc fraction, MeOH fraction and n-BuOH fraction from raw walnut was $95.14\pm0.36\%,\; 90.02\pm1.19\%\;and\;89.41\pm0.81\%$, respectively. The tert-butylhydroperoxide (t-BHP) treatment in vitro increased lactate dehydrogenase release and lipid peroxidation in renal cortical slices. Such changes were completely prevented by addition of MeOH fraction, EtOAc fraction and n-BuOH fraction of walnut. These results indicate that the walnut extract exerts the benedicial effect against t-BHP-induced cell injury and its effect may be due to antioxidant action. In addition, it is suggested that walnut extract might be developed as the effective scavenger for the prevention of oxidative stress.
과산화수소로 유도된 SH-SY5Y 신경세포 사멸에 대한 오미자·칠해목 추출혼합물의 보호효과
박은국,한경훈,이승희,김남기,배문형,서영하,용윤중,정선용,최춘환,Park, Eun-kuk,Han, Kyung-Hoon,Lee, Seung-Hee,Kim, Nam-Ki,Bae, Mun-Hyoung,Seo, Young-Ha,Yong, Yoon-joong,Jeong, Seon-Yong,Choi, Chun-Whan 한국식품영양학회 2018 韓國食品營養學會誌 Vol.31 No.6
In neuronal cell deaths, oxidative stress is normally implicated with a most of these deaths occurring in neurodegenerative disorders such as the Alzheimer's and Parkinson's diseases. In this study, the neuroprotective effects of Schisandra chinensis (SC) and Ribes fasciculatum (RF) extracts on hydrogen peroxide ($H_2O_2$)-induced oxidative stress in neuroblastic cell line were investigated. For an hour, hydrogen peroxide of $100{\mu}M$ concentration, was induced on neuroblastic cells, causing apoptic cell death. For the neuroprotection, a sample of neuroblastic cells had been pre-treated with SC and RF extracts for 24 hours before application of the hydrogen peroxide. No neurotoxic effects were observed in the cells that had been treated by SC and RF. This prove that the treatment of SC and RF extract prevented apoptotic cell death of neuroblastic cell line exposed to oxidative injury. In addition, applying both SC and RF extracts at a 7:3 ratio increased the neuronal cell survival rate, compared to individual treatments of SC and RF extract. This study suggests that SC and RF extracts may be potential therapeutic agents for the prevention of neuronal cell death.
오미자칠해목 추출물의 과산화수소로 유발된 PC12뇌세포 사멸과 스코폴라민으로 유발된 렛드 동물모델에 대한 개선 효과
박은국,한경훈,허재혁,김남기,배문형,서영하,용윤중,정선용,최춘환,Park, Eun-kuk,Han, Kyung-Hoon,Heo, Jae-Hyeok,Kim, Nam-Ki,Bae, Mun-Hyoung,Seo, Young-Ha,Yong, Yoon-joong,Jeong, Seon-Yong,Choi, Chun-Whan 한국식품영양학회 2020 韓國食品營養學會誌 Vol.33 No.3
Cognitive impairment is considered to be key research topics in the field of neurodegenerative diseases and in understanding of learning and memory. In the present study, we investigated neuroprotective effects of Schisandra chinensis (SC) and Ribes fasciculatum (RF) extracts in hydrogen peroxide-induced neuronal cell death in vitro and scopolamine-induced cognitive impairment in Sprague Dawley<sup>®</sup> (SD) rat in vivo. Apoptotic cell death in neuroblastic PC12 cell line was induced by hydrogen peroxide for 1 hour at 100 μM. However, mixture of SC and RF treatment prevented peroxide induced PC12 cell death with no neurotoxic effects. For in vivo experiment, the effect of SC and RF extracts on scopolamine-induced cognitive impairment in SD rat was evaluated by spontaneous alternation behavior in Y-Maze test. After 30 min scopolamine injection, the scopolamine-induced rats presented significantly decreased % spontaneous alteration and acetylcholine level, compared to non-induced group. However, treatment of SC+RF extracts rescued the reduced % spontaneous alteration with acetylcholine concentration from hippocampus in scopolamine-induced rats. These results suggested that mixture of SC and RF extract may be a potential natural therapeutic agent for the prevention of cognitive impairment.