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RAW 264.7 대식세포에 미치는 병꽃나무 꽃 추출물의 항산화 및 항염증 효과
유영춘(Yung Choon Yoo),이계원(Gye Won Lee),조영호(Young Ho Cho) 한국생명과학회 2016 생명과학회지 Vol.26 No.3
본 연구에서는 병꽃나무 꽃 추출물의 항산화 효과 및 LPS로 자극된 RAW 264.7 대식세포에서 항염증 효과를 조사하였다. 병꽃나무 꽃 추출물의 총 폴리페놀과 플라보노이드 함량을 측정한 결과 총 폴리페놀 함량과 플라보노이 함량은 각각 719.19±0.04 ㎍/ml, 644.87±0.02 ㎍/ml로 나타났다. 항산화 활성을 확인하기 위하여 DPPH 라디칼과 superoxide 음이온 라디칼 소거 활성을 측정한 결과 투여 농도 의존적으로 항산화 활성이 증가하는 것으로 나타났다. 또한 항염증 활성을 확인하기 위하여 염증 매개물질인 NO와 inflammatory cytokine인 IL-6와 TNF-α의 생성량을 측정하였다. 그 결과 NO와 IL-6의 생성을 투여농도 의존적으로 저해하였지만, TNF-α의 경우 병꽃나무 꽃 추출물의 농도가 20 ㎍/ml 이하에서는 TNF-α의 생성을 억제하지 않았고 100 ㎍/ml 이상에서는 오히려 분비를 유도하는 것으로 나타났다. 또한, 병꽃나무 꽃 추출물의 염증반응과 관련된 iNOS 발현과 MAPK 및 NF-κB의 활성에 미치는 영향을 측정한 결과 iNOS는 농도 유의적으로 그 발현이 감소되었으며, MAPK의 발현 및 인산화에는 별다른 영향을 미치지 못하는 것으로 나타났다. 반면에 IκB의 인산화를 효과적으로 감소시켜 NF-κB의 활성을 억제하여 항염증 활성을 나타내는 것을 확인할 수 있었다. 이상의 결과로부터 병꽃나무 꽃 추출물은 항산화 및 항염증 활성이 우수한 기능성 소재로 활용이 가능할 것으로 사료된다. This study investigated the antioxidant and anti-inflammatory activity of ethanol extract from the flowers of Weigela subsessilis (WS-E) in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. The total polyphenol and flavonoid content was 719.19±0.04 ㎍ tannic acid equivalents/ml and 644.87±0.02 ㎍ quercetin equivalents/ml, respectively. The antioxidant activities of WS-E were measured by 1,1-diphenyl-2-picrylhydrazyl (DPPH) and superoxide anion radical scavenging activity. The antioxidant activities of WS-E increased markedly, in a dose-dependent manner. To screen for anti-inflammatory agents, the inhibitory effects of WS-E on the production of proinflammatory cytokines in the LPS-stimulated RAW 264.7 macrophages was examined. WS-E had no effect on cell viability at a concentration of 100 μg/ml. Nitric oxide (NO) and interleukin (IL)-6 production were inhibited in a dose-dependent manner (p<0.05). WS-E had no effect on the production of tumor necrosis factor (TNF)-α at a concentration of 0.16–20 μg/ml but induced TNF-α at a concentration of 100 ㎍/ml. Inducible nitric oxide synthase (iNOS) expression was also inhibited at lower concentrations (p<0.05). In addition, WS-E reduced the activation of nuclear factor (NF)-κB by inhibition of inhibitoy (I) κB phosphorylation in RAW 264.7 macrophages upon stimulation with LPS (100 ng/ml) for 24 h but not that of mitogen-activated protein kinase (MAPK). These results suggest that WS-E may be a useful antioxidant and anti-inflammatory agent in functional cosmetics.
마우스 Macrophage의 IL-1 및 TNF-${\alpha}$의 분비유도에 있어서 한국산 겨우살이 추출물이 미치는 영향
윤택준,유영춘,홍은경,조영호,이석원,유보림,김종배,Yoon, Taek-Joon,Yoo, Yung-Choon,Hong, Eun-Kyung,Cho, Young-Ho,Lee, Suk-Won,Azuma, I.,Yoo, Bo-Im,Kim, Jong-Bae 한국생약학회 1994 생약학회지 Vol.25 No.2
To investigate the effect of Korean mistletoe on stimulation of macrophage, the activity to induce interleukine-1(IL-1) and tumor necrosis factors-${\alpha}(TNF-{\alpha})$ from murine peritoneal macrophage by its extracts originated from oak was examined. From in vitro analysis of the cytokines using the culture supernatants of macrophages stimulated with its extracts for 1hr, it was found that Korean mistletoe induces IL-1 and $TNF-{\alpha}$ from murine macrophage. Furthermore, both extracts of Korean mistletoes that were extracted with distilled water and 2% acetic acid exhibited a significant activity to induce two cytokines. In the stimulation for 30 min, Korean mistletoe at concentration of $1{\sim}100\;\mu/ml$ showed a significant induction of IL-1 from macrophage until 24 hrs after stimulation, showing maximal activity on $5{\sim}10\;hrs\;at\;10{\sim}100\;\mu/ml$. On the other hand, $TNF-{\alpha}$ was induced on the early period, 2 hrs, after stimulation at a wide range of concentration, $1{\sim}500\;\mu/ml$. In addition, the fraction of Korean mistletoe from 80% saturated ammonium sulphate precipitation showed a significant activity to induce both cytokines from macrophage. The present study demonstrates that Korean mistletoe contains immunoregulatory factors responsible for stimulating murine macrophage to secrete IL-1 and $TNF-{\alpha}$ which play an important role in immune responses, and suggests that the activity of Korean mistletoe to induce two cytokines is functioned by a possible independent stimulation manner.
한국산 겨우살이 추출물(KME)의 2형 당뇨 억제 및 근육세포 미토콘드리아 생성 증가 효과
정회윤(Hoe-Yune Jung),유영춘(Yung Choon Yoo),김인보(Inbo Kim),성낙윤(Nak Yun Sung),최옥병(Ok-Byung Choi),최보화(Bo-Hwa Choi),김종배(Jong-Bae Kim) 한국식품영양과학회 2015 한국식품영양과학회지 Vol.44 No.3
본 연구에서는 C57BL/6J ob/ob 마우스를 이용하여 한국산 겨우살이 냉수 추출물(KME)의 항당뇨 활성을 조사하였다. 50 혹은 100 mg/kg의 KME를 1일 1회씩 경구투여 한 결과 KME 투여 개시 5일 후부터 ob/ob 마우스의 혈당이 유의하게 억제되었으며, 10일 후부터 안정된 억제 효과를 나타내고 대조군에 비해 20% 이상의 혈당강하 효과를 나타내었다. 경구 당부하 실험(OGTT)에서는 KME 경구투여 마우스에서 유효한 당부하 억제 활성이 관찰되었다. 또한 KME 경구투여는 ob/ob 당뇨 마우스의 혈액 내 총 콜레스테롤과 중성지질의 농도를 억제하는 것으로 나타났다. 한편 C2C12 근육세포를 이용한 in vitro 실험에서 KME를 처리함으로써 glucose uptake가 현저히 증가하였다. 한편 매우 흥미롭게도 KME를 처리한 C2C12 근육세포에 있어서 미토콘드리아 생성과 산화대사 조절물질인 peroxisome proliferatoractivated receptor gamma coactivator 1-α(PGC-1α)를 비롯하여 glucose transporter type 4(GLUT4), estrogen-related receptor-α(ERR-α), nuclear respiratory factor-1(NRF-1) 그리고 mitochondrial transcription factor A(TmfA)와 같은 PGC-1α 관련 유전자들의 발현이 증가하는 것으로 확인되었다. 이 결과는 KME가 2형 당뇨에 대한 치료물질로서의 작용을 지니며 이러한 KME의 항당뇨 활성은 미토콘드리아 생성의 조절과 관련 있는 것으로 추정된다. In this study, the anti-diabetic activity of a cold water extract of Korean mistletoe (KME) was investigated in C57BL/6J Lep ob (ob/ob) mice. Oral administration of KME (50 or 100 mg/kg/d) significantly inhibited the level of blood glucose of ob/ob mice after 5 days from the beginning of KME treatment. And the anti-diabetic effect of KME was stabilized 10 days after oral administration, showing a substantial reduction of blood glucose levels by more than 20% as compared with control mice. The results of oral glucose tolerance test (OGTT) revealed that oral administration of KME gave rise to a remarkable improvement in overall glucose response. Oral administration of KME in ob/ob diabetic mice also significantly reduced blood total cholesterol (TCHO) and triglyceride (TG) levels compared with the diabetic control mice. Moreover, in an in vitro experiment using C2C12 myotubes, treatment of KME prominently increased glucose uptake. Interestingly, KME significantly increased the expression of peroxisome proliferator-activated receptor gamma coactivator 1-α (PGC-1α), a head regulator of mitochondrial biogenesis and oxidative metabolism, and PGC-1α-associated genes such as glucose transporter type 4 (GLUT4), estrogen-related receptor-α (ERR-α), nuclear respiratory factor-1 (NRF-1), and mitochondrial transcription factor A (TmfA) in C2C12 cells. These results suggest that KME has potential as a novel therapeutic agent for diabetes, and its anti-diabetic activity may be related to the regulation of mitochondrial biogenesis.
배종환(Jong Hwan Bae),유영춘(Yung Choon Yoo),홍장희(Jang Hee Hong),문은호(Eunho L. Moon),송경식(Kyung Sik Song),이경복(Kyung Bok Lee) 한국실험동물학회 2004 Laboratory Animal Research Vol.20 No.1
A modification of a colorimetric assay was used to determine the concentration of total and individual sulfated-glycosaminoglycan (GAG) in equine synovial fluid and serum with joint disease. For the identification of enzymatic active products of the equine synovial fluid, strong anion exchange-high performance liquid chromatography (SAX-HPLC) was performed. By the action of chondroitin ABC lyase, three unsaturated disaccharides, 2-acetamide-2-deoxy-3-O-(β-D-gluco-4-enepyranosyluronic acid)-D-galactose (△Di-COS), 2-acetamide-2- deoxy-3-O-(β-D-gluco-4-enepyranosyluronic acid)-6-O-sulfo-D-galactose (△Di-C6S), and 2-acetamide-2-deoxy-3-O-(β-D-gluco-4-enepyranosyluronic acid)-4-O-sulfo-D-galactose (△Di-C4S), were produced from the equine synovial fluid. Total GAG concentration of the equine synovial fluid was 0.82± 0.24 ㎎/㎖ (mean±SD) in normal horses, but it was decreased in horses with joint disease (0.17±0.12 ㎎/㎖). The concentration of keratan sulfate (KS), chondroitin sulfate (CS), and hyaluronic acid (HA) were decreased in horses with joint disease. The concentration of serum HA of normal horses was 77.00±66.14 ㎍/㎖, but it was markedly increased in joint diseases (168.50±147.50 ㎍/㎖), There appears to be some correlation between joint inflammation and serum HA levels as determined by experimental studies of animals. Irrespective of the explanation, it is clear that measurement of the level of serum HA may provide a useful marker for monitoring the onset and progression of a number of important diseases and disorders in equine.