cAMP 증가 유도 약물의 대식세포- 및 T 세포-매개성 면역반응 조절작용

이만휘,조재열,Rhee, Man-Hee,Cho, Jae-Youl 대한약학회 2007 약학회지 Vol.51 No.1

To investigate the immunomodulatory roles of cyclic AMP (CAMP) on macrophage- and T lymphocyte-mediated immune responses, CAMP elevating agents were employed and carefully re-examined under the activation conditions of the cells. Various inhibitors tested dose-dependently blocked tumor necrosis factor (TNF)-${\alpha}$ production with IC$_{50}$ values ranged from 0.04 to 300 ${\mu}$M. Of the inhibitors, cAMP-elevating agents showed lower cytotoxicity assessed by lactate dehydrogenase (LDH) release, suggesting less toxic and more selective. In particular co-treatment of dbcAMP with a protein kinase C inhibitor staurosporine displayed the synergistic inhibition of TNF-${\alpha}$ production. The modulatory effect of dbcAMP on TNF-${\alpha}$ and nitric oxide (NO) was significantly affected by treatment time of dbcAMP. Thus, post-treatment of dbcAMP (three hours before LPS) abrogated dbcAMP's inhibitory activity and rather enhanced TNF-${\alpha}$ level up to 60%. In contrast, additional NO production was shown at the co-treatment of dbcAMP with LPS. Unlike simultaneous treatment of phorbol 12-myristate 13-acetate (PMA) and interferon (IFN)-${\gamma}$co-treatment, the combination of dbcAMP with other NO-inducing stimuli did not show drastic overproduction of NO. cAMP elevating agents also diminished splenocyte proliferation stimulated by concanavalin (Con) A, phytohemaglutinin A (PHA) and lipopolysaccharide (LPS). In addition, dbcAMP but not rolipram strongly suppressed CD8$^+$ T cells (CTLL-2). Finally, cAMP elevating agents were differentially involved in regulating CD98-mediated cell-cell adhesion. Thus, dbcAMP and rolipram significantly enhanced the cell-cell adhesion, whereas forskolin blocked. Therefore, our results suggest that CAMP elevating agents participate in various immune responses mediated by macrophages and T cells with a different fashion depending on cellular environments and activation signals.

Anti-Platelet Function of the Non-Saponin Fraction from Panax ginseng C.A. Meyer (Korean red ginseng)

박화진,이만휘,박경미,남기열,박기현,Park, Hwa-Jin,Rhee, Man-Hee,Park, Kyeong-Mee,Nam, Ki-Yeul,Park, Ki-Hyun 생화학분자생물학회 1993 한국생화학회지 Vol.26 No.8

고려홍삼의 비사포닌분획(NSF)은 높은 농도의 트롬빈(0.2~$2{\mu}/ml$)에 의해 유도된 사람혈소판 응집반응을 강하게 억제했다. 그 응집정도는 $Ca^{2+}$ 길항제인 verapamil의 응집억제 정도보다 강했다. Verapamil은 혈소판 응집반응을 저해시키는 혈소판내 cAMP의 생성을 증가시켰지만, NSF는 cAMP의 생성증가에는 현저한 영향이 없었다. NSF는 $TXA_2$의 생성과 serotonin의 방출을 억제시키고 있었다. NSF는 혈소판막 인지질로부터 arachidonic acid의 방출을 억제하지 않고, arachidonic acid를 $TXA_2$로 전환시키는 어떤 경로를 차단하므로 혈전중의 치료 및 예방에 중요한 역할을 할 것으로 추측된다. The non-saponin fraction(NSF: lipophilic fraction) from Korean red ginseng showed an inhibitory activity in the aggregation of human platelets induced by a high dose of thrombin (0.2 to 2 u/ml), the extent of which was stronger than that of verapamil, a $Ca^{2+}$ antagonist. NSF did not affect the elevation of the intracellular cAMP concentration which inhibits platelet aggregation, as shown in the case of verapamil, but inhibited the production of $TXA_2$ and the release of serotonin. NSF does not inhibit the release of arachidonic acid from membrane phospholipids, but inhibits a certain pathway transforming arachidonic acid to $TXA_2$. NSF may play a critical role in preventing and curing thrombotic diseases.

Panax ginseng C. A . Meyer ( 고려홍삼 ) 의 비사포닌의 분획의 항혈소판작용

박화진,이만휘,박경미,남기열,박기현 ( Hwa Jin Park,Man Hee Rhee,Kyeong Mee Park,Ki Yeul Nam,Ki Hyun Park ) 생화학분자생물학회 1993 BMB Reports Vol.26 No.8

The non-saponin fraction(NSF: lipophilic fraction) from Korean red ginseng showed an inhibitory activity in the aggregation of human platelets induced by a high dose of thrombin (0.2 to 2 u/㎖), the extent of which was stronger than that of verapamil, a Ca^(2+) antagonist. NSF did not affect the elevation of the intracellular cAMP concentration which inhibits platelet aggregation, as shown in the case of verapamil, but inhibited the production of TXA₂ and the release of serotonin. NSF does not inhibit the release of arachidonic acid from membrane phospholipids, but inhibits a certain pathway transforming arachidonic acid to TXA₂. NSF may play a critical role in preventing and curing thrombotic diseases.

인삼의 혈압조절 작용에 대한 최근 연구동향

이휘민,이만휘,Lee, Hui-Min,Rhee, Man-Hee 고려인삼학회 2007 고려인삼연구와 산업 Vol.1 No.1

기와층버섯 추출물의 항비만활성, 항암활성 및 단회경구독성시험

강은희(Eun-Hee Kang),이인경(In-Kyoung Lee),황미현(Mi-Hyun Hwang),최재영(Jae-Young Choi),창즐치앙(Zhi-Qiang Chang),이만휘(Man-Hee Rhee),윤봉식(Bong-Sik Yun),강성철(Cheng-Zhe Jiang),김길수(Kil-Soo Kim),박승춘(Seung-Chun Park) 한국독성학회 2007 Toxicological Research Vol.23 No.3

In this study, we investigated the in vitro anti-obesity, anti-cancer activity and single oral dose toxicity of Inonotus xeranticus extracted by methanol (INXM) or ethyl acetate (INXE). In order to investigate anti-obestity effect of Inonotus xeranticus extracts, the 3T3-L1 cells were treated with these extracts at various concentrations(1, 10, 100 and 300 ㎍/㎖). It was observed that 3T3-L1 cells treated with 100 ㎍/㎖ of Inonotus obliquus ethyl acetate extract (INOE), INXM and INXE, in the absence of differentiation cocktail (0.5 mM isobutylmethylxanthine (IBMX) 1 μM dexamethasone, 1 ㎍/㎖ insulin), differentiated at a rate of 78.5, 80.9, and 76.4% respectively. Differentiation rates of 86.6% and 83.4% were observed in 3T3-L1 cells which were treated with differentiation cocktail at 100 ㎍/㎖ of INXM and INXE, respectively. The anti-cancer effect of Inonotus xeranticus extracts was investigated using a method of sulforhodamine B in sarcoma 180 cell line. The cells were treated with these extracts (1, 10, 100 and 300 ㎍/㎖) for 48 hours. The growth of cells which were treated with 300 ㎍/㎖ of INXM was inhibited by 80.1%. The growth of sarcoma 180 cells which were treated with 100 and 300 ㎍/㎖ of INXE was inhibited by 74.7% and 64.5%, respectively. In single oral dose toxicity study, no differences were observed between control and treated groups in clinical signs, body weight gains, and feed and water consumptions. The results indicated that Inonotus xeranticus extracts did not show any toxic effects at 2,000 ㎎/㎏ in mice, and the LD?? of these extracts was found to be higher than 2,000 ㎎/㎏ in this experiment. From the above results, Inonotus xeranticus methanol and ethyl acetate extracts might have useful clinical applications in the management of cancer and obesity and may also be useful as a medicinal food.

Phellinus linteus, Phellinus baumii 및 Phellinus gilvus의 형태, 일반성분, 원소분석 및 무기성분 함량의 비교

배재성,장광호,이만휘,정규식,조우식,최성국,김영환,박승춘,Bae, Jae-sung,Jang, Kwang-ho,Rhee, Man-hee,Jeong, Kyu-shik,Jo, Woo-sik,Choi, Sung-guk,Kim, Young-hoan,Park, Seung-chun 대한수의학회 2003 大韓獸醫學會誌 Vol.43 No.3

진흙버섯 품종별 P. linteus, P. baumii 그리고 P. gilvus에 대한 형태학적 차이, 일반성분 및 무기성분을 조사하였다. 전자현미경 사진에서 P. gilvus와 P. linteus는 비슷한 형태를 보여주고 있다. 원소 함량의 구성비에서는 세 종류의 버섯이 모두 유사함을 보여주었다. 일반성분을 분석 비교시 수분의 함량은 P. baumii가 가장 높았으며 P. gilvus의 수분 함량은 10% 이내로 낮은 경향을 보여주었다. 식이섬유는 약 30% 이상을 함유하고 있으나 vitamin C 및 단백질의 함량은 높지 않았다. 원소분석에서 Ca의 경우 세 종류의 진흙버섯에서 P. baumi가 가장 높은 Ca 함량을 보여주었으나 통계적유의성은 없었다. 그러나 원소조성 및 일반성분 비교에서는 P. linteus, P. baumii 그리고 P. gilvus의 큰 차이점이 나타나지 않았다. The purpose of this study is to compare the morphology, general composition, elemental composition and mineral contents of Phellinus linteus, Phellinus baumii and Phellinus gilvus. In the scanning electron microscopy, P. gilvus and P. linteus had a similar shape. In the elemental composition, all of the three Phellinus spp. showed similar percent for the analyzed components. In the general composition, the water content of P. linteus. P. baumii and P. gilvus were 13%, 15% and 10%, respectively. The dietary fiber of three Phellius spp. showed more than 30%. The range of vitamin C and protein content was 1.5-2.1 g% and 3.63-3.73 g%. respectively. In mineral analysis, P. baumii of the three Phellinus spp. has the highest calcium concentration (1,135 ppm). From the above results, P. linteus. P. baumii and P. gilvus did not show any differences in the general composition and elemental composition.

pH 및 고온 열처리가 홍삼물추출물의 주종 사포닌 성분변화에 미치는 영향

최금희(Keum-Hee Choi),곽이성(Yi-Seong Kwak),이만휘(Man-Hee Rhee),황미선(Mi-Sun Hwang),김석창(Seok-Chang Kim),박채규(Chae-Kyu Park),한경호(Gyeong-Ho Han),송경빈(Kyung-Bin Song) 고려인삼학회 2008 Journal of Ginseng Research Vol.32 No.2

홍삼 물 추출물의 사포닌 품질안정성을 조사할 목적으로 100℃이상의 고온으로 열처리한 후 추출물의 사포닌 성분변화를 조사하였다. 홍삼 물 추출물의 pH을 산성(pH 3.0), 중성 (pH 7.0), 알카리성(pH 10.0)으로 조정 하여 100, 110, 120℃에서 30분간 열처리한 후 사포닌성분을 조사한 결과 대조군에 비해 모든 처리군에서 감소하는 경향을 나타내었다. 특히 pH 3.0의 산성조건에서는 사포닌 분해 현상이 TLC에서 관찰되었고 HPLC로 함량을 분석한 결과도 total ginsenoside 함량이 대조군의 1.99에 비해 100℃에서는 1.22, 110℃에서는 1.05, 120℃에서는 0.97로 현저히 감소하는 경향을 나타내었다. 또한 각 pH 별로 조정한 후 100, 110, 120℃로 열처리한 엑기스의 PD/PT 사포닌 비율도 1.33~1.47 수준으로 대조군의 1.89에 비해 크게 감소하는 경향을 나타내었다. 그러나 이러한 열처리한 물 추출물에서의 PD/PT 감소비율은 pH 및 온도 별로 큰 차이가 나타나지 않는 경향이었다. 대조군에 비해 열처리한 추출물의 PD/PT 사포닌 비율감소로 볼 때 ginsenoside-Re, -Rgl 등의 PT계 사포닌 보다는 ginsenoside-Rb₁, -Rb₂, -Rc, -Rd등의 PD계 사포닌이 100℃ 이상의 고온에서 열에 더욱 불안정한 것으로 나타났다. This study was carried out to investigate the changes of ginsenosides composition in Korean red ginseng water extract (RGWE) after heated with high temperatures above 100℃. RGWEs were adjusted with pH 3.0, pH 7.0 and pH 10.0, respectively, and then heated at 100, 110 and 120℃ for 30 minutes by using autoclave. Total ginsenosides of RGWE treated with heating showed decreasing tendency when compared with control. By TLC analysis, decreasing effect of ginsenosides in RGWE were significantly observed in the acidic condition of pH 3.0, particulary. By HPLC analysis, total ginsenoside of control showed 1.89%, while those of RGWE treated with 100, 110 and 120℃ showed 1.22, 1.05 and 0.97%, respectively. The ratio of protopanaxadiol (PD) to protopanaxatriol (PI) saponins in control was 1.89, while that of PD/PT in treated RGWEs were level of 1.33 to 1.47. By the result of decreased ratio of PD/PT in RGWE, it was considered that PD type saponin such as ginsenoside-Rb₁, -Rb₂, -Rc and -Rd was more unstable than PT type saponin such as ginsenoside-Re and Rg against high temperature heating above 100℃.

고온 열처리가 홍삼물추출물의 이화학적 특성에 미치는 영향

곽이성(Yi-Seong Kwak),최금희(Keum-Hee Choi),경종수(Jong-Soo Kyung),원준연(Jun-Yeon Won),이만휘(Man-Hee Rhee),이재곤(Jae-Gon Lee),황미선(Mi-Sun Hwang),김석창(Seok-Chang Kim),박채규(Chae-Kyu Park),송경빈(Kyung-Bin Song),한경호(Gyeong-H 고려인삼학회 2008 Journal of Ginseng Research Vol.32 No.2

홍삼추출물의 품질안정성을 조사할 목적으로 100℃이상의 고온으로 장시간 열처리한 후 추출물의 이화학적 특성변화를 조사하였다. 홍삼물추출물의 pH를 조정하지 않고 100, 110, 120℃에서 열처리 시간을 2시간까지 연장하여 학대조건으로 처리한 후 사포닌, 유리당, 색도 및 무기물 등의 이화학적성분 변화를 조사하였다. 사포닌의 total ginsenoside 함량은 대조군 1.99에 비해 100, 110, 120℃에서 각각 1.65, 1.45, 1.29%로 온도가 증가함에 따라 온도 의존적으로 감소하는 경향이었다. 고온으로 열처리한 물추출물의 사포닌 안정성을 좀더 자세히 조사하기 위해 열에 안정한 홍삼특유 사포닌인 ginsenoside-Rh₂ 및 -Rg₃를 TLC로 조사한 결과 다른 ginsenoside가 감소되는 것과는 반대로 대조군에 비해 110, 120℃로 처리한 엑기스에서 비교적 강한 spot이 관찰되었다. 유리당 함량은 고온 열처리한 엑기스에서 frutose, glucose, maltose의 함량이 증가하는 경향을 나타내었으며 무기물함량은 주요한 무기성분 Fe, P, Ca, K, Mg중 Fe, Ca, Mg이온 이 대조군에 비해 큰 폭으로 감소하는 경향을 나타내었다. 고온 열처리된 홍삼물추출물의 색도를 조사한 결과 명도 L값은 대조군과 거의 대등한 수준이었으나 적색도 a 값은 대조군의 -0.86에서 100, 110, 120℃ 열처리시 +0.04, +0.05, +1.14로 증가하는 경향이었고 황색도 b값도 대조군의 27.68에서 각각 33.61, 33.61, 37.42로 증가하는 경향을 나타내었다. 총 색상인 색차(△E) 값은 대조군의 0.23에서 각각 6.08, 6.04, 11.74로 증가하는 경향을 나타내었고 100℃ 및 110℃보다는 120℃에서 증가폭이 큼을 알 수 었었다. 따라서 고온으로 열 처리시 a, b 값이 증가되어 전체적인 총 색상(△E)값이 증가됨으로 100이상의 고온으로 물추출물을 열처리시 갈색화 반응이 많이 진행됨을 알 수 있었다. This study was carried out to investigate the some physicochemical properties of Korean red ginseng (Panax ginseng C.A. Meyer) water extract (RGWE) after heated with high temperatures above 100℃ for 2 hours. RGWEs were heated at 100, 110 and 120℃ for 2 hours by using autoclave. After RGWEs were heated at high temperature for 2 hours without not adjustment of pH, the changes of saponin, free sugars, mineral and color in the RGWEs were investigated. Total ginsenoside content in control was 1.99%, while those of RGWE were 1.65, 1.49 and 1.29% when treated at 100, 110 and 120℃, respectively. The contents of total ginsenoside showed decreased tendency as heating temperatures were increased. The ginsenoside-Rh₂ and -Rg₃, which have been reported as very stable red ginseng ginsenosides, showed relatively strong spots on TLC when RGWEs were heated at 110 and 120℃. In case of free sugars in RGWEs, fructose, glucose and maltose showed high contents when compared with control, while Fe, Ca and Mg ions showed very low contents. Value of L in RGWE treated with high temperature was almost the same with control, while values of a and b were increased. Values of a were increased from -0.86 of control to +0.04, +0.05 and +1.14 when treated with 100, 110 and 120℃, respectively. Values of b also were increased from 27.68 of control to 33.61, 33.61 and 37.42 when treated with 100, 110 and 120℃, respectively. Values of total color in RGWEs treated with high temperatures, E, were finally increased by values of a and b.

쌀겨발효추출물의 항혈전효과

전보라,지현동,김수정,이천휘,김태완,이만휘,Jeon, Bo-Ra,Ji, Hyun Dong,Kim, Su Jung,Lee, Chun-Hee,Kim, Tae-Wan,Rhee, Man-Hee 대한수의학회 2015 大韓獸醫學會誌 Vol.55 No.4

Although the effects of the rice bran have recently been investigated, there is no information regarding platelet physiology available. However, it is well known that fermented natural plants have a beneficial effect on cardiovascular diseases. Therefore, this study was conducted to investigate whether fermented rice bran extract (FRBE) with several plants (Artemisia princeps, Angelica Gigantis Radix, Cnidium officinale, and Camellia sinensis) affected agonist-induced platelet aggregation, and if so, what the underlying mechanism of its activity was. We performed several experiments, including in vitro platelet aggregation, intracellular calcium concentration and adenosine triphosphate release. In addition, the activation of integrin ${\alpha}_{II}b{\beta}3$ was determined using fibrinogen binding. Thrombus formation was also evaluated in vivo using an arterio-venous shunt model. The FRBE inhibited collagen-induced platelet aggregation in a concentration-dependent manner. FRBE significantly and dose dependently attenuated thrombus formation using rat arterio-venous shunt. FRBE suppressed the intracellular calcium mobilization in collagen-stimulated platelets. We also found that FRBE inhibited extracellular stimuli-responsive kinase 1/2, p38-mitogen-activated protein kinases and c-Jun N-terminal kinase phosphorylation. These results suggested that FRBE inhibited collagen-induced platelet aggregation, which was mediated by modulation of downstream signaling molecules. In conclusion, FRBE could be developed as a functional food against aberrant platelet activation-related cardiovascular diseases.

마우스 회장 운동에 대한 나문재 (Suaeda asparagoides M_IQ) 추출물의 효과

송재찬,박창희,김현탁,메하리 엔델,이만휘,박승춘,김길수,김태완,Song, Jae-Chan,Park, Chang-Hee,Kim, Hyun-Tak,Endale, Mehari,Rhee, Man-Hee,Park, Seung-Chun,Kim, Kil-Soo,Kim, Tae-Wan 대한수의학회 2006 大韓獸醫學會誌 Vol.46 No.4

Suaeda (S.) asparagoides $M_{IQ}$, one of the halophyte groups, has been used as a folk remedy for digestive disturbances in Korea. However, its pharmacological activity on gastrointestinal motility has not been reported yet. In this study, the effects of this halophyte extracts with various solvent fractions (ethanol, hexane, chloroform, ethyl acetate, butanol, and water) on mice ileal spontaneous motility was examined. All solvent fractions at the concentration of $100{\mu}g/ml$ showed inhibitory actions on spontaneous motility of ileum with the potency order of water > 70% ethanol > hexane ${\gg}$ chloroform ${\geq}$ butanol ${\geq}$ ethyl acetate, respectively. In addition, the water fraction of extracts from S. asparagoides $M_{IQ}$ (WFSA) dose-dependently ($1-100{\mu}g/ml$) inhibited the amplitude of spontaneous phasic contraction and area under the contractile curve (AUC). The inhibitory effect of water fraction at the concentration of $10{\mu}g/ml$ was not affected by tetrodotoxin (TTX), $Na^+$ channel blocker ($1{\mu}M$), and $N^w$-nitro-L-arginine Methyl Ester (L-NAME), nitric oxide synthase inhibitor ($100{\mu}M$). However, cyclopiazonic acid (CPA, $10{\mu}M$), inhibitor of sarcoplasmic reticulum $Ca^{2+}$-ATPase, almost blocked the inhibitory effects of WFSA ($10{\mu}g/ml$) on the spontaneous phasic contraction of mouse ileum. But, CPA did not inhibit the lowering basal tone effects of WFSA. The result of this study showed that various extracts of S. asparagoides $M_{IQ}$ induce inhibitory effects on spontaneous contraction of mice ileal segments. More over, the polar solvent fractions were shown to be more potent than non-polar solvent fractions. The effects of S. asparagoides $M_{IQ}$ extracts are not mediated by nerve or nitric oxide. The inhibitory effects of WFSA at least partially mediated by sarcoplasmic reticulum $Ca^{2+}$-ATPase. However, further study is required to determine the exact pharmacological mechanisms of this halophyte on its gastrointestinal motility inhibitory effects.