국내산 밤 일부 품종의 기능성 성분분석과 항산화 및 대식세포 활성

이현주(Hyeon-Ju Lee),정미자(Mi Ja Chung),조재열(Jae Youl Cho),함승시(Seung-Shi Ham),최면(Myeon Choe) 한국식품영양과학회 2008 한국식품영양과학회지 Vol.37 No.9

국내산 밤 일부 품종(단택, 대보, 석추, 옥광, 병고)의 기능성 성분 분석과 이들 물질들의 항산화력 및 밤 추출물이 대식세포 활성에 미치는 영향력을 알아보았다. 국내산 밤 일부품종의 내피와 과육에서 coumarin, gallic acid 그리고 catechin이 HPLC에 의해 검출되었다. 밤 내피에 가장 풍부한 기능성 성분은 catechin이고 그 다음 gallic acid와 coumarin 순이었으나, 과육에서는 gallic acid만 검출되었다. 따라서 밤의 기능성 성분인 gallic acid와 catechin의 항산화력을 DPPH radical 소거작용과 SOD 유사활성을 통해 알아보았다. Gallic acid과 catechin을 6.0 mg/100 g 처리하였을 때 DPPH radical 소거작용이 각각 69.4%와 38.3%였으며, gallic acid의 DPPH radical 소거작용은 농도 의존적이었다. Gallic acid를 동일한 농도의 catechin과 비교하면 모든 농도구간에서 catechin보다 gallic acid가 높은 DPPH radical 소거활성을 나타내었다. 그러나 SOD 유사활성은 gallic acid보다 catechin이 높았다. 더하여 밤의 과육, 노란과육 및 흰과육 추출물들이 대식세포 활성에 미치는 영향력을 알아보았다. 병고 흰과육 추출물은 현저하게 대식세포 활성을 증가시켰고, 병고, 단택, 대보 및 옥광의 노란과육 추출물 역시 아무것도 처리하지 않은 세포와 비교하여 현저히 대식세포를 활성화시켰다. 노란과육 추출물이 흰과육 추출물보다 더욱 더대식세포 활성을 증가시켰고, 5품종 중 병고가 대식세포 활성을 가장 증가시켰다. Antioxidative and macrophage phagocytic activities and contents of functional component in selected Korean chestnuts (Dantaek, Daebo, Okkwang, Seokchu, Byunggo) were evaluated. Coumarin, gallic acid and catechin in inner skin and whole kernel of selected Korean chestnuts were detected by HPLC. The predominant functional components in inner skin of chestnut were catechin, followed by gallic acid and coumarin. However, the whole kernel had only gallic acid. Thus, the antioxidant properties of gallic acid and catechin were evaluated through DPPH radical-scavenging activity and SOD like activity. Gallic acid and catechin at 6.0 mg/100 g exhibited 69.4% and 38.3% of scavenging activities on DPPH radical, respectively. DPPH radical-scavenging activity of gallic acid increased in a concentration-dependent manner. Gallic acid was found to possess higher DPPH radical-scavenging activity than equivalent amount of catechin at all concentrations, whereas catechin was found to have higher SOD like activity than gallic acid. In addition, pre-incubation of macrophage with white kernel extract from Byunggo resulted in a significant increase of phagocytic activity and yellow kernel extracts from Byunggo, Dantaek, Daebo and Okkwang, leading to an increase in phagocytic activity compared with untreated cells. Yellow kernel extracts was found to have higher phagocytic activity than white kernel extracts. Byunggo had the highest phagocytic activity. The results suggest that the Korean chestnut may provide a natural source of antioxidants and active immunity.

Effects of Atropine, Phentolamine and Propranolol on Calcium uptake, Superoxide generation and Phagocytic activity in activated PMN Leukocytes

이정수,한은숙,이광수,Lee, Chung-Soo,Han, Eun-Sook,Lee, Kwang-Soo The Korean Society of Pharmacology 1988 대한약리학잡지 Vol.24 No.1

세포질 내 칼슘 농도의 증가는 다형핵 백혈구의 산화성 대사를 자극하는 주요 인자로 여겨지고 있다. 활성화된 다형핵 백혈구로부터 lysosomal enzyme의 유리는 세포내 cyclic nucleotide농도에 따라 조절될 수 있지만 신경전달물질과 ${\beta}$-아드레날린 또는 무스카린성 수용체의 결합에 따른 그밖의 반응은 아직도 분명하지 않다. 덧붙여, ${\alpha}$-아드레날린성 수용체의 중개에 의한 다형핵 백혈구의 기능은 알려져 있지 않다. Atropine, phentolamine과 propranolol은 활성화된 다형핵 백혈구의 칼슘흡수, superoxide 생성, NADPH oxidase 활성도 그리고 식작용을 억제하였으며, 이에 반하여 carbachol과 isoproterenol은 활성화된 세포의 반응을 약간 더 자극하였다. Carbachol또는 isoproterenol 의하여 항진된 superoxide 생성은 각각 그들의 길항제인 atropine과 propranolol 의하여 억제되었다. 활성화된 다형핵 백혈구의 반응은 chlorpromazine, verapamil과 dantrolene에 의하여 억제되었으나 lithium에 의 하여 약긴 항진되었다. 한편 chlorpromazine과 dibucaine은 NADPH oxidase 활성도에 영향을 주지 않았다. Atropine, phentolamine과 propranolol은 칼슘에 의존적인 식작용을 억제 하였다. 이상의 결과로부터 atropine, phentolamine과 propranolol은 칼슘 유입을 억제하고 자율 신경계의 수용체와 연관이 있는 NADPH oxidase계에 직접 작용함으로써 활성화된 다형핵 백혈구로부터 superoxide 생성을 억제할 것으로 시사되었다. Although the release of lysosomal enzymes from activated PMN leukocyte can be regulated by intracellular cyclic nucleotide levels, other responses of PMN leukocyte according to the binding of neurotransmitters to either ${\beta}$-adrenergic or muscarinic receptors are still not clarified. In addition, the function of PMN leukocyte mediated by ${\alpha}$-adrenergic receptors is uncertain. Atropine, phentolamine and propranolol inhibited calcium uptake, superoxide generation, NADPH oxidase activity and phagocytic activity in activated PMN leukocyte, whereas carbachol and isoproterenol slightly further stimulated the responses of activated cells. Either carbachol or isoproterenol stimulated superoxide generation was inhibited by their antagonists, atropine and propranolol, respectively. The response of activated PMN leukocyte was inhibited by chlorpromazine, verapamil and dantrolene but slightly stimulated by lithium. On the other hand, chlorpromazine and dibucaine did not affect NADPH oxidase activity. Atropine, phentolamine and propranolol suppressed the calcium dependent phagocytic activity. Thus, the results suggest that atropine, phentolamine and propranolol may inhibit superoxide generation in activated PMN leukocyte by the inhibition of calcium influx and by their direct action on the NADPH oxidase system which is associated with autonomic receptors.

靑皮가 마우스의 免疫細胞에 미치는 影響

殷載淳,吳贊鎬,鄭鉉雨 대한동의병리학회 1998 동의생리병리학회지 Vol.12 No.2

靑皮는 疏肝破氣 및 消積化滯하는 작용이 있는 약물로 대부분 氣滯로 인한 증상들에 사용되고 있지만 임상상 活血之劑와 配合하여 氣滯血瘀나 血瘀脇痛등에 應用되고 있다. 氣滯는 각종의 스트레스등으로 인하여 발생되어 血行을 不暢하게 하고, 심지어는 正氣까지도 손상시킨다. 正氣의 손상은 바로 免疫機能의 低下라고도 말할 수 있는데, 免疫이란 非自己로 인식된 異常細胞를 처리함으로써 個體의 恒常性을 維持하려는 현상을 말한다. 면역에 先天的 免疫과 T림프구 및 B림프구가 관여하는 特異性 免疫이 있고, 免疫作用에 관여하는 細胞로는 特異性 免疫에 관여하는 免疫細胞外에도 Macrophage, NK cell, 수지상 세포 및 랑게르한스 세포등을 들 수 있다. 뇌의 기능은 심장에서 방출되는 혈액의 공급에 의해 이루어지는데, 만약 혈액의 공급이 제대로 이루어지지 않으면 많은 질환들이 발생한다. 그 중 虛血性 腦疾患은 충분한 양의 혈액이 뇌를 공급되지 않거나 공급되고 있는 혈액내의 산소농도가 떨어져 발생하는 것으로써 임상적으로는 運動麻痺·知覺痲痺등 神經機能 消失등의 증상을 보이기 때문에 東醫學的으로는 이를 風病이라 하고, 風病은 대체적으로 陰虛나 血虛등으로도 발생할 수 있어 陰虛動風證 또는 血虛生風證등의 病證들이 있다. 그리하여 각종 氣滯로 인한 氣滯則血滯가 발생되거나 또는 正氣가 손상되어 氣虛則血虛하기 때문에 陰虛나 血虛등으로 인한 運動麻痺나 知覺麻痺가 발생한다. 이러한 이유등으로 本 硏究者들은 혈류장애 질환치료에 사용되는 韓藥材中 靑皮의 작용을 규명하는 硏究의 일환으로 靑皮가 면역기능에 어떠한 영향을 미치는가에 대하여 알아보기 위하여 동결건조된 靑皮를 마우스에 1일 1회씩 7일간 투여한 후 T-lymphcyte의 Apoptosis와 sub-population, 그리고 proliferation 및 Peritoneal macrophage의 nitric oxide 생성과 phagocytic activity를 관찰한 결과 DNA fragmentation과 sub-population에는 별다른 영향을 주지 않았지만 proliferation에 있어서는 증식이 촉진되었고, 복강 macrophage의 nitric oxide 생성과 phagocytic activity에 있어서는 NO는 감소하였으나 phagocytic activity는 현저히 증가하였다. 이는 靑皮가 NO 생성을 억제하여 phagocytic activity를 증가시킨 결과라 思料된다. 이상과 같은 實驗結果들을 통해서 볼 때 靑皮는 臨床的으로 각종 氣滯로 인한 正氣損傷과 血液循環障碍가 동시에 나타날 때에만 正氣를 보충해주는 役割을 할 수 있을 것으로 사료된다. The purpose of this research was to investigate effects of Citri Reticulatae Viride Pericarpium water extract(CRVP) on immunocytes in mice. The administration of CRVP did not affect apoptosis and sub-population of mouse T-lymphocytes, but increased the proliferation of T-lymphocytes. CRVP decreased nitric oxide production from mouse peritoneal macrophage, but increased a phagocytic activity of macrophages. These results suggest that CRVP does not affect T cell mediated immunity, but increases the phagocytic activity of peritoneal macrophages.

$\beta$-glucan을 접종한 한국산 메기(Silurus asotus)의 호중구와 리소자임 활성 증강

박성우,김영길,최동림,Park, Sung-Woo,Kim, Young-Gill,Choi, Dong-Lim 한국어병학회 1996 한국어병학회지 Vol.9 No.1

한국산 메기(Silurus asotus)에 $\beta$-glucan을 복강내에 접종한 후, 경시적으로 말초혈액중의 호중구의 수와 PAS반응의 변화, 호중구의 탐식능 및 혈청중의 리소자임의 활성변화를 지표로 비특이적방어능력의 증강유무를 조사하였다. 글루칸의 투여후 말초혈액중의 호중구수를 증가시켰으며 투여회수에 따라 2회 접종한 경우 호중구의 증가가 지속되는 기간이 1회 접종어보다 길게 나타났다. 그러나 호중구의 PAS반응에는 변화가 없었다. 또 탐식율도 현저하게 증가하였지만 탐식지수는 변화가 없었다. 리소자임의 활성은 2회 접종시만이 현저히 증가하였으나 1회 접종시에는 활성의 증가가 없었다. 그러므로 $\beta$-glucan의 투여는 말초혈액중의 호중구수와 탐식능의 증가 및 리소자임 활성의 증가등 비특이적방어인자의 활성을 증가시켜 어체를 방어하는 능력을 증강시키는 것임을 알 수 있다. To examine the effectiveness of $\beta$-glucan in non-specific defence mecanisms of Korean catfish(Silurus asotus), the activities of peripheral neutrophils and serum lysozyme were evaluated after once or twice $\beta$-glucan injections. The number of peripheral neutrophils increased after the injection. The increased number in neutrophils in twice $\beta$-glucan injections maintained longer than in single glucan injecton. But the neutrophils did not showed any remarkable changes in PAS-positive reaction. Peripheral neutrophils from fish injected glucan had enhanced phagocytic activities but there was no significant difference in phagocytic index for the glucan and control groups. An elevation of lysozome activity was also observed in the fish received twice glucan but neither single glucan or physiological saline injection could not evoked an increase of lysozyme activity. These results suggest that $\beta$-glucan can enhance the resistance of Korean catfish against to bacteria infection through the activation of peripheral neutrophils and lysozyme.

운목향 70% 에탄올 추출물의 항염증 및 면역조절에 대한 효과

김민선,김남석,권진,김하림,이다영,오미진,김홍준,이창현,오찬호 한국약용작물학회 2018 한국약용작물학회지 Vol.26 No.1

Background: This present study was conducted to evaluate the anti-inflammatory and immune regulatory effects of Aucklandia lappa Decne (AL). Methods and Results: We measured cytotoxicity, nitric oxide (NO) content, mRNA expression (iNOS, IL-1α, IL-1β, and TNF-α), protein expression (iNOS, COX-2, and IκB-α) and phagocytic activity in RAW264.7 cells. Male BALB/c mice were fed 100 ㎎/㎏ AL (Aucklandia lappa Decneon 70% ethanol extract) and 250 ㎎/㎏ AL for 4 weeks; thereafter, we observed B/T or CD4+/CD8+ lymphocyte subpopulation change, and expression patterns of CD4+ and CD8+ lymphocytes by immunohistochemical staining in mouse splenocytes and/or thymocytes. To determine the experimental concentration of AL, cell viability was measured by MTT assay and tested at 12.5 ㎍/㎖ or less. AL inhibited the levels of NO, lymphokine production (IL-1β, and TNF-α), and mRNA (iNOS, IL-1α, IL-1β, and TNF-α) and protein (iNOS, and COX-2) expression. Additionally, the levels of IκB-α, phagocytic activity, and splenic and thymic T lymphocytes, especially TH and TC cells were significantly increased in AL administered mice. The immuno-reactive density of CD4+ and CD8+ lymphocytes was stronger in AL groups than in the normal group. AL stimulated NO, iNOS, and COX-2, and regulated IL-1α, IL-1β, TNF-α, and IκB-α in macrophages treated with LPS (lipopolysaccharide). In addition, AL increased the phagocytic activity of macrophages and the immunity of mouse T (TH, and TC) cells. Conclusions: These results suggested that AL might show anti-inflammatory activity via the suppression of various inflammatory markers and immuno-regulatory activity.

청피(靑皮)가 뇌혈류와 복강내 Macrophage에 미치는 영향

김남순 ( Nam Soon Kim ),황경택 ( Kyung Taek Whang ),황일택 ( Il Taek Whang ),강덕수 ( Duk Soo Kang ),강학천 ( Hak Chun Kang ),한종현 ( Jong Hyun Han ) 대한경락경혈학회 2001 Korean Journal of Acupuncture Vol.18 No.2

Citri Reticulatae Viride Pericarpium is being used to regulate the flow of gi. However, the mechanism of its pharmacological actions is not well understood. The purpose of this research was to investigate effects of Citri Reticulatae Viride Pericarpium on peritoneal macrophages in mice and blood pressure, regional cerebral blood flow in rats. Nitric oxide production was tested using a Griess reagents, and a phagocytic activity of mouse peritoneal macrophage was tested using a luminometer. Blood pressure and regional cerebral blood flow was tested using a phygiograph, MacLab and Macintosh computer with data acquisition system. 1. Blood pressure was increased by Citri Reticulatae Viride Pericarpium in a dose-dependent manner. 2. rCBF was increased by Citri Reticulatae Viride Pericarpium in a dose-dependent manner. 3. Pretreatment with propranolol significantly inhibited Citri Reticulatae Viride Pericarpium induced increase in rCBF. 4. Citri Reticulatae Viride Pericarpium inhibited nitric oxide production from peritoneal macrophages in mice. 5. Citri Reticulatae Viride Pericarpium accelerated phagocytic activity of peritoneal macrophage in mice These results suggest that Citri Reticulatae Viride Pericarpium causes a diverse response of blood pressure, regional cerebral blood flow(rCBF), nitric oxide production and phagocytic activity. The increase in rCBF is also mediated by adrenergic βrecepter.

베타-카로틴의 면역생물학적 연구

안영근,구자돈,김정훈,김봉희,조필형,구교임 충남대학교 약학대학 의약품개발연구소 1992 藥學論文集 Vol.8 No.-

Effects of beta-carotene on the immunobiological responses were studied in ICR mice. ICR male mice were divided into 8 groups (10 mice/group), and beta-carotene at doses of 4, 20 and 100 ㎎/㎏ were orally administered to ICR mice once daily for 28 consecutive days. Cyclophosphamide (CY) was injected intraperitoneally (i.p.) to ICR mice with a single dose of 5 ㎎/㎏ body weight at 2 days before secondary immunization. Mice were sensitized and challenged with sheep red blood cells (S-RBC). Immune responses were evaluated by humoral immunity, cellular immunity and non-specific immunity. The results of this study were summarized as follows; (1) Beta-carotene significantly increased the weight ratios of liver, spleen and thymus to body weight depending on dose, and significantly increased the increasing rate of body weight and the number of circulating leukocyte. (2) Beta-carotene dose-dependently increased hemagglutination titer, Arthus reaction and hemolytic plaque forming cell related to humoral immunity. (3)Beta-carotene significantly increased delayed-type hypersensitivity reaction and rosette forming cell relatted to cellular immunity. (4) Beta-carotene dose-dependently increased phagocytic activity, and significantly increased natural killer (NK) cell activity. (5) Beta-carotene dose-dependently inhibited reductions in humoral immunity, cellular immunity, NK cell activity and phagocytic activity by treatment with CY.

Production of Phagocyte Activating Supernatants by Olive Flounder (Paralichthys olivaceus) Leucocytes Stimulated with Genomic DNA of Escherichia coli

Lee Chan Hwei,Kim Dong Soo,Kim Ki Hong The Korean Society of Fisheries and Aquatic Scienc 2002 Fisheries and Aquatic Sciences Vol.5 No.4

Effects of Escherichia coli genomic DNA on the production of phagocyte activating supernatants by the head kidney leucocytes isolated from olive flounder (Paralichthys olivaceus) were investigated. Phagocyte activating activity of the supernatants was estimated by. measuring reactive oxygen species (ROS) production in target head kidney phagocytes. All supernatants from olive flounder head kidney leucocytes-stimulated with E. coli DNA induced significantly (P<0.01) higher ROS production from target pagocytes than the unstimulated control supernatant. Maximum enhancement of chemiluminescent response was observed $5.0-10.0{\mu}g\;mL^{-1}$ of bacterial DNA while the increment ability was decreased significantly (P<0.01) at the concentration of $20.0{\mu}mL^{-1}$. The results demonstrate that olive flounder head-kidney leucocytes stimulated with bacterial DNA release a soluble phagocyte activating cytokines capable of enhancing the respiratory burst activity from target phagocytes.

Euonymus alatus추출물의 실험적 당뇨발생 억제효과

김태중,송희종,소준노,이정호 大韓免疫學會 1996 大韓免疫學會誌 Vol.18 No.4

Insulin-dependent diabetes mellitus (IDDM) is majorly caused by an immune system-mediated destruction of the beta cells in the islets of Langerhans. Patients with IDDM face major changes in lifestyle and the possibility of debilitating and life-threatening complications. In Korean traditional medicine, Euonymus alatus (EA) extract has been widely used for preventing and curing ulcer and inflammation. A previous study has shown that EA itself has an antidotic activities against inflammation, suggesting possibility that EA can exert this beneficial effects to IDDM by an initial protection against diabetes caused by pancreatic inflammation. The present study was undertaken to evaluate the prophylactic effect of EA extract on the diabetogenesis of streptozotocin (STZ) and alloxan (ALX), using animal model, and to explore the possible mechanisms of its antidiabetic activity. Multiple low doses of STZ (45 mg/Kg for 5 days) in ICR mice and single dose of ALX (60 mg/Kg) in SD rats induced high level of hyperglycemia (defined as plasma glucose level > 300 mg/dl) and remarkably diminished body weight. But, animals that received diabetogenic agents in combination withsEA were maintained plasma glucose level as a normal state ( < 150 mg/dl), and appeared in good health and had similar weights to those of normal control. These results strongly indicated that EA prevented and blocked diabetogenesis of chemicals. EA significantly inhibited the nitric oxide- and tumor necrosis factor-production of mouse peritoneal macrophages. However, EA enhanced interleukin 1 production of macrophages at lower concentration but not at higher. EA itself increased phagocytic activity of unstimulated monocytes, but it did completely block that activity of zymosan- or platelet activating factor-stimulated cells. Taken together, this study led to conclusion that EA itself has a significant prophylactic effects against diabetes, and also suggested that its antidiabetic effects might be manifested by the downregulation of inflammatory process in diabetogenesis.

Quercetin이 마우스의 면역반응에 미치는 영향

안영근,박영길,김정훈 원광대학교 식품약품안전성연구소 1991 食品藥品安全性硏究 Vol.4 No.-

Effects of quercetin on the specific and non-specific immune responses were studied in vivo. Quercetin at a dose of 2.5, 5, 10, 20 and 40 ㎎/㎏ were orally administered to ICR male mice once daily for 28 consecutive days. Cuclophosphamide was injected intraperitoneally to ICR mice with a single dose of 5㎎/㎏ 2 days before secondary immunization. Mice were sensitized and challenged with sheep red blood cells (S-RBC). Immune responses were evaluated by humoral and cellular immune reponses and non-specific immune response. The results of this study were summerized as followings: 1. Quercetin significantly decreased the body weight, and introduced the atrophy of liver, spleen and thymus gland dose-dependently, but increased the numbers of white blood cell. 2. Querectin significantly depressed the hemagglutination titer, Arthus reaction and hemolythtic plaque forming cell. 3. Querectin significantly depressed the delayed type hypersensitivity and rosette forming cell. 4. Quercetin at a dose of 2.5, 5 and 40㎎/㎏ significantly depressed phagocytic activity. 5. Quercetin at d dose of 10 and 20 ㎎/㎏ significantly increased natural killer cell activity.