염증은 외부 자극으로부터 보호하는 중요한 면역반응이다(Jeong et al., 2012). 그러나 NO 및 inflammatory cytokine과 같은 염증 매개 인자의 과도한 생성은 비정상적인 염증 반응을 초래할 수 있다(Paradise et al., 2010). 염증 매개 인자의 생성과 염증 신호 전달을 조절하는 중요한 역할을 하는 대식세포는 LPS 자극에 의해 NF-κB가 활성화 되어 inflammatory cytokine 등의 염증 매개 인자들이 분비가 증가되며 염증 반응을 심화시킨다(Lee et al., 2012). 본 연구에서는 영지버섯균 발효 꾸지뽕나무 가지 톱밥 추출물을 이용하여 LPS로 염증을 유도한 Raw264.7세포에서 염증 관련 인자들의 생성 및 발현에 미치는 영향을 분석하여 항염증 효과를 확인하였다. NO는 염증에서 중요한 역할을 하며 대식세포의 염증 반응 조절 평가시 대표적인 지표로 사용되며 iNOS에 의해 생성이 유도되며 PGE2는 염증 매개 인자로 inflammatory cytokine 생성에 관여하며 COX-2에 의해 생성이 유도된다 (Paradise et al., 2010; Posadas et al., 2000). 발효추출물 이 RAW264.7세포에서 세포독성 없이 NO의 생성과 PGE2의 생성을 감소시켰으며 이 결과는 발효 추출물 처리에 의해 iNOS와 COX-2의 발현이 감소한 것과 일치하 였다. 또한 염증 반응을 조절하는 대표적인 inflammatory cytokine으로 알려진 IL-1β, TNF-α의 생성이 감소되었다. 따라서 발효추출물은 NO, PGE2, inflammatory cytokines 생성을 감소시켜 항염증 효과를 나타낸다고 볼 수 있다. NF-κB는 iNOS, COX-2 및 inflammatory cytokine의 발현을 조절하는 전사인자로 IκB와의 결합에 의해 NF-κB의 핵 내 이동이 억제되며 불활성화상태로 존재한다. LPS 자극에 의해 IκB가 인산화 및 분해되면 NF-κB가 활성화 되어 핵 내로 이동하여 염증성 매개인자들의 발현을 유도 하고, 염증성 질환 뿐만 아니라 다양한 질환을 유발시키는 것으로 알려져 있다 (Kawai and Akira, 2006; Ghosh and Ksarin, 2002). 본 연구에서는 LPS에 의해 자극된 RAW264.7 세포에서 IκB의 분해 및 NF-κB의 전이가 발 효추출물에 의해 감소됨을 확인하였으며 발효추출물에 의한 iNOS, COX-2 및 inflammatory cytokine 감소는 NF- κB 활성화 감소에 의해 조절되는 것으로 사료된다. 이러한 결과들을 통해서 발효추출물이 NF-kB 활성화 억제를 통해 염증 매개 인자들의 생성 및 발현을 감소시킴으로서 염증 반응 억제 효과를 나타냄을 확인하여, 염증 관련 질환에 대해 예방 및 개선을 위한 항염증 기능성 소재로 사용될 수 있음을 시사한다.
Home-made soy sauces with or without Hovenia dulcis Thunb (Hutgae) originated from different parts such as fruits, stems, and twigs were prepared according to the Korean traditional procedure. Soy sauces supplemented with Hutgae were evaluated for their activities of 2,2-diphenyl-1-picrylhydrazyl radical scavenging (DPPH) and alcohol dehydrogenase (ADH), free amino acid profiles, and sensory quality. All soy sauce types containing Hutgae had a strong DPPH activity as compared to the general type of soy sauce without Hutgae (GSC). Among Hutgae groups, DPPH activities of soy sauce supplemented with Hutgae stems was higher than that of soy sauces with either Hutgae fruits or twigs. ADH activities of soy sauces with Hutgae ranged from 14% to 55%, thus indicating that the functional activity of Hutgae was not altered during soy sauce preparations. Total free amino acid content of GSC was 295.5 ㎎%, and that of soy sauce with Hutgae fruits (346.8 ㎎%) was the highest when compared to Hutgae stems (272.3 ㎎%) and Hutgae twigs (225.6 ㎎%). In amino acid profiles, aspartate, arginine, histidine, and lysine levels were higher in soy sauces with Hutgae compared to GSC, whereas isoleucine, leucine, and phenylalanine levels were lower. Particularly, high levels of aspartate, glutamate, threonine, and lysine were presented in Hutgae twigs, whereas for Hutgae fruits and Hutgae stems, the levels of serine, glycine and arginine, and proline and methionine were high, respectively. According to sensory evaluations, Hutgae stems were preferred than GSC, due to the lower offensive smell and higher umami tastes. These findings demonstrate that soy sauce with Hutgae stems has potential protective effects against hangovers, improves the taste, and implies a possible functional ingredient.
In this study, the antioxidative effects and inhibitory effects on tyrosinase and elastase of Sorbus commixta (S. commixta) twig extracts were investigated. The aglycone fraction of S. commixta twig extract showed the prominent free radical (1,1-diphenyl-2-picrylhydrazyl, DPPH) scavenging activity(FSC50, 13μg/mL). Reactive oxygen species (ROS) scavenging activities of S. commixta twig extracts on ROS generated in Fe3+-EDTA/H2O2 system were investigated by the luminol-dependent chemiluminescence assay. The 50 % ethanol extract among extracts showed the most prominent ROS scavenging activity (OSC50, 0.189μg/mL). The cellular protective effects of extract/fractions of S. commixta twig on the rose-bengal sensitized photohemolysis of human erythrocytes were investigated. The 50 % ethanol extract and ethyl acetate fraction showed the cellular protective effects against ROS in a concentration dependent manner (5~50μg/mL). The inhibitory effect of S. commixta twig extract on tyrosinase was investigated to assess the whitening efficacy. The ethyl acetate (IC50, 113.2μg/mL) and aglycone fraction(IC50, 105.3μg/mL) on tyrosinase showed more remarkable inhibitory effect than arbutin(IC50, 226.88μg/mL), known as the whitening agent. The inhibitory effect of aglycone fraction (IC50, 6.9μg/mL) on elastase was simillar to quercetin(IC50, 6.1μg/mL), flavonoid known as reference compound. These results indicate that extract/fractions of S. commixta twig can function as antioxidants in biological systems, particularly skin exposed to UV radiation by scavenging 1O2 and other ROS, and protect cellular membranes against ROS. S. commixta twig extracts can be applicable to new functional cosmetics for anti-aging products.
Pine wood nematode (PWN), Bursaphelenchus xylophilus is associated with the pine wilt disease and transmitted by pine sawyer, Monochamus alternatus. Because pine sawyer has one-year life cycle, one natural infection of PWN is occurred a year. Therefore, artificial propagation method of PWN is needed to improve experiment associated with PWN. In this study, effect of diameter, paraffin sealing of twig and dosage on pine wood nematode reproduction in Japanese black pine, Pinus thunbergii. PWN reproduction was compared in twigs of P. thunbergii and P. densiflora. Numbers of reproduced PWN were higher with decreasing diameter of twig. Distance (5 and 10 cm) from inoculation site of PWN did not influence reproduction of PWN. Reproduced numbers of PWN were higher in the paraffin-sealing twig than non-sealing twig. Dosage of PWN influenced reproduction of PWN. Reproduction rate was the highest at the rate of 10 IJs (13.7 and 61.1-fold increasing in P. densiflora and P. thunbergii, respectively 30 days later) whereas lowest at the rate of 1000 Ijs (1.1 and 0.7-fold increasing in P. densiflora and P. thunbergii, respectively 30 days later). Numbers of reproduced PWN were more in P. thunbergii than P. densiflora.
Background : Phenolic compounds were isolated from the twig of Broussoneita Kazinoki. Their structures were established on the basis of extensive spectroscopic (MS, 1D , and 2D NMR) data analysis and by comparison with the spectroscopic data reported in the literature. Methods and Results : The twig of B. Kazinoki were extracted 60% aqueous ethanol for 3 days at room temperature. The extract was filtered and concentrated by vacuum evaporator. And then, extract was partitioned using hexane, methylene chloride (MC), ethyl acetate (EtOAc), n-butyl alcohol (BuOH) and H2O, successively. The extraction was separated by using prep-HPLC, and the structure was analyzed by Mass spectrometry (MS) and 1H-, 13C-, 1H-1H COSY, HSQC, HMBC NMR data. Conclusion : These compounds were identified as chlorogenic acid (1), ferulic acid (2), p-coumaric acid (3), taxifolin (4), marmesin (5), 5-methoxy marmesin (6), pinoresinol (7), syringaresinol (8), quercetin (9), broussonin A (10), broussonin B (11), broussoflavonol A (12), broussoflavonol B (13), kazinol A (14), and 5,7,3',4'-tetrahydroxy-3-methoxy-8,5'-diprenylflavone (15).
As a result of cytotoxic compounds against cancer cell lines from natural sources, senven compounds were isolated from the leaf and twig of Acer okamotoanum Nakai. The compounds (1-7) were identified as ethyl gallate (1), methyl gallate (2), gallic acid (3), trans resveratrol-3-O-β-D-glucopyranoside (4), acertannin (5), nikoenoside (6), and fraxin (7) by physicochemical and spectroscopic data (including mp, UV, IR, MS, 1H-NMR, 13C-NMR, DEPT, and HMBC) in comparison with those of published papers. All the compounds were tested for their cytotoxic activity against L1210, HL-60, K562, and B16F10 cancer cell lines in vitro by MTT assay method. Compounds 1-3 and 5 showed cytotoxic activity against all tested cell lines with IC50 values ranged from 12.5 to 72.2 μM. Of the compounds, methyl gallate (2) exhibited the most potent cytotoxic activity against L1210, HL-60, K562, and B16F10 tumor cells with IC50 values of 12.5, 48.3, 22.8, and 22.8 μM, respectively. Other compounds did not show any cytotoxic activity against four cancer cell lines.