Streptococcus mutans로부터 분리한 Mutan이 흰쥐의 골결손부 치유에 미치는 영향

권현정 ( Hyun Jung Kwon ),김용현 ( Yong Hyun Kim ),한국일 ( Kook Il Han ),정의길 ( Eui Gil Jung ),한만덕 ( Man Deuk Han ) 한국치위생과학회 2015 치위생과학회지 Vol.15 No.2

We investigated the effects of a mutan (water-insoluble a-glucans) isolated from Streptococcus mutans on the healing of bone defect in rat. Sprague-Dawley rats were divided into control (saline-treated), lipopolysaccharide (LPS)-treated, and mutan-treated groups (n=6 per group). Experimental bone defects were surgically created with round fissure bur at the buccal surface of the left mandibular. The control groups was administered with saline solution (0.1 ml/100 g), while the LPS and mutan group was given LPS and mutan (1 mg/kg body weight) three times weekly. After 4 weeks the rats were sacrificed, the healing of bone defect was assessed by bone mineral density (BMD) and micro-computed tomography (mCT) examination. Percent bone volume (bone volume/tissue volume [BV/TV]), trabecular thickness (Tb.Th), and trabecular number (Tb.N) parameters of mCT showed higher values in control group than LPS and mutan group. Bone surface/volume ratio (BS/BV), trabecular bone pattern factor (Tb.Pf), and structure model index parameters of mCT showed higher values in LPS group than mutan group. BMD values of mutan treated-alveolar bones were significantly lower for than that of the LPS group. Therefore, we suggest that mutan, water-insoluble a-glucans from S. mutans may be induce the induction of periodontal diseases.

광귤나무(Citrus aurantium) 열매에서 분리된 쿠마린의 iNOS 억제효과

김범선(Beom sun Kim),정의길(Eui Gil Jung),윤형선(Hyung Sun Youn),황경아(Kyung A Hwang),한만덕(Man Deuk Han),유연호(Yon ho Yu),남궁우(Kung Woo Nam) 대한약학회 2017 약학회지 Vol.61 No.5

Citrus fruits have been used as an edible fruit, and also as a traditional medicine, since ancient times. A crude methanol extract of Citrus aurantium was shown to suppress inducible proinflammatory cytokines, but its constituents were not known. In this study, we isolated four furanocoumarins, bergapten (1), imperatorin (2), isoimperatorin(3), and phellopterin (4), and one 7-geranyloxycoumarin, auraptene (5), from a methanol extract of dried Citrus aurantium fruit. The structures of these compounds were elucidated by extensive application of spectroscopic techniques, including 2D NMR spectroscopy and mass spectrometry. All of the isolated compounds were evaluated for their ability to inhibit the production of proinflammatory cytokine inducible nitric oxide synthase (iNOS) in lipopolysaccharide-activated RAW264.7 cells. Compounds 2−5 significantly suppressed iNOS production but compound 1 had no iNOS inhibitory effects. These results indicate that at least three furanocoumarins and one 7-geranyloxycoumarin are involved in the suppressive effects of citrus fruits against the production of inducible inflammatory cytokines.

Streptococcus dysgalactiae로부터 분리된 히알루론산과 황화된 유도체의 구조와 항염증 활성

홍창일(Chang-Il Hong),정의길(Eui-Gil Jung),한국일(Kook-Il Han),김용현(Yong Hyun Kim),이성희(Sung Hee Lee),이홍섭(Hong Sub Lee),한만덕(Man-Deuk Han) 한국생명과학회 2016 생명과학회지 Vol.26 No.5

히알루론산(HA, Hyaluronic acid)은 β-1, 3-N-acetyl glucosamine과 β-1, 4-glucuronic acid가 반복된 선형 폴리머 고분자로서 생물학적 활성 및 생체친화성 특성 때문에 의약 및 약학분야에서 중요한 분자로 여겨지고 있다. 본 연구는 HA을 S. dysgalactiae으로 얻고, 화학적 방법을 통해 황화된 히알루론산(S-HA, Sulfated hyaluronic acid)유도체를 합성하여 그 구조와 항염증 활성을 비교하였다. HA의 생산은 S. dysgalactiae를 5 l 생물반응기를 이용하여 대량 배양하여 수용성 히알루론산(HA-WS, water soluble hyaluronic acid)과 비수용성 히알루론산(HA-WI, water insoluble hyaluronic acid)을 분리 정제하였다. 특히 HA-WI를 황화시켜 황화된 히알루론산(S-HA) 유도체를 합성하였으며, 그 수율은 90%로 나타났다. 합성된 S-HA의 구조를 FT-IR 및 ¹H/<SUP>13</SUP>C-NMR를 통해 S. dysgalactiae 로부터 생산된 표준 HA, HA-WS 및 HA-WI와 비교 분석한 결과, 황으로 치환된 양상을 확인하였다. 또한, S-HA의 항염증 활성을 RAW 264.7 대식세포를 통해 확인한 결과, S-HA는 천연 형태의 HA (HA, HA-WS)보다 nitric oxide (NO)와 COX-2 및 PGE₂ 유전자 발현이 유의하게 낮게 발현되었다. 염증 매개 cytokine인 TNF-α (<80 pg/ml) 및 IL-6 (<100 pg/ml)의 생성도 S-HA가 천연 HA보다 낮은 수준으로 정량되었다. 이 같은 결과에서 황화된 S-HA은 천연 히알루론산보다 용해성이 우수하고 염증관련 사이토카인의 생성 억제를 통해 항염증 효과를 나타내므로 염증치료제, 성형 및 생체 적용 약물전달 소재로 그 활용이 기대된다. Hyaluronic acid (HA) is an important macromolecule in medical and pharmaceutical fields. HA is a natural and linear polymer composed of repeating disaccharide units of β-1, 3-N-acetyl glucosamine and β-1, 4-glucuronic acid. This work aimed to confirm the structural characteristics and anti-inflammatory activities of HA and its chemically sulfated-HA. HA was produced from a fed-batch fermentation process using Streptococcus dysgalactiae in a 5 l bioreactor. HA was isolated water-soluble form (HA-WS) and water-insoluble form (HA-WI) from culture medium, and was obtained chemically sulfated-derivative (S-HA) that resulted in a 90% yield from HA-WI. The structural features of the sulfated- HA (S-HA) were investigated by FT-IR and ¹H-NMR spectroscopy. The FT-IR and NMR patterns revealed the similarity in both the FTIR spectrum as well as NMR spectrum of both reference standard and purified HA from S. dysgalactiae. The anti-inflammatory activities of HA and S-HA were examined on LPS-induced RAW 264.7 cells. S-HA was significantly inhibited production of pro-inflammatory mediators such as nitric oxide (NO) and PGE₂ and the gene levels of iNOS and COX-2, which are responsible for the production of NO and PGE₂, respectively. Furthermore, S-HA also suppressed the overproduction of pro-inflammatory cytokine TNF-α (<80 pg/ml) and IL-6 (<100 pg/ml) compared to that of HA-WI. The present study clearly demonstrates that HA-S exhibits anti-inflammatory activities in RAW 264.7 macrophage cells.

고지방식이로 고지혈증이 유도된 흰쥐에서 황화된 수용성 β-glucan의 항고지혈증 효과

김용현(Yong Hyun Kim),한국일(Kook-Il Han),전미애(Miae Jeon),황선구(Seon Gu Hwang),정의길(Eui-Gil Jung),권현정(Hyun-Jung Kwon),한만덕(Man-Deuk Han) 한국생명과학회 2014 생명과학회지 Vol.24 No.11

본 연구는 영지버섯 균사체로부터 얻은 수용성 베타글루칸(SBG)이 고지방식이로 유도한 흰쥐의 항고지혈 효과 및 혈중 지질수준에 미치는 영향을 조사하였다. 5주령의 수컷 흰쥐에 고지방식이를 2주간 급여하여 비만을 유도하고, 그 후 2주 동안 다음과 같은 5가지 group의 식이를 급여하였다: 1) 일반사료 식이군(NC), 2) 고지방사료식이군(HC), 3) 고지방사료 및 고농도(200 mg/kg) SBG 식이군(HC-HSBG) 4) 고지방사료 및 저농도(20 mg/kg) SBG 식이군(HC-LSBG), 5) 고지방사료 및 로바스타틴(lovastatin) 식이군(HC-Lov). 실험 결과, 식이 효율(Food efficiency ratio)은 고지방식이를 급여한 군들에서 일반사료 식이군에 비하여 증가하였으나 통계적 유의성은 없었다. 혈액학적 결과에서 고농도 및 저농도의 SBG와 로바스타틴 투여군은 고지방식이만 급여한 군(HC)보다 호중구와 중성구 수치가 유의성 있게 증가하였다. 또한 혈청 내 총콜레스테롤, 중성지질 및 LDL-C는 고농도 및 저농도 SBG 투여군 모두에서 유의성 있게 감소되었다. 이 같은 결과는 화학적으로 황화된 수용성 베타글루칸(SBG)이 흰쥐에서 혈중 콜레스테롤 수치를 낮추어 항고지혈증 효과를 나타내는 것을 의미한다. The aim of this study was to investigate the effect of water-soluble sulfated β-glucan (SBG) obtained from Ganoderma lucidum mycelia on the antihyperlipidemic and serum lipid levels in high-fat diet-induced obese rats. Five-week-old male Sprague-Dawley rats were fed a high-fat diet for two weeks to induce obesity. They were ten divided into five groups-normal control diet group (NC), high-fat control diet group (HC), high-fat diet and 200 mg/kg of SBG group (HC-HSBG), high-fat diet and 20 mg/kg of SBG group (HC-LSBG), and high-fat diet and 20 mg/kg of lovastatin group (HC-Lov)-and fed one of five diets for two more weeks. Although food intake and final body weight after four weeks of SBG consumption were similar in the five experimental groups, food efficiency ratio was higher in the high-fat diet groups(2, 3, 4, and 5) than in the NC group. In evaluating the hematological parameters of the rats, the neutrophil and monocyte ratios were higher in the HC-HSBG, HC-LSBG, and HC-Lov groups than in the HC group. Serum lipid profiles were analyzed after a 12 hr fast at the end of the study. Total cholesterol, triglyceride, and low-density lipoprotein cholesterol (LDL-C) levels were significantly lower in the HC-HSBG and HC-LSBG groups than in the HC group. These results suggest that chemically engineered sulfated mushroom β-glucan (SBG) might contribute to lower cholesterol and lipid levels in blood.

호두과피 추출물의 항산화 및 항균활성

한국일(Kook-Il Han),김미란(Mi ran Kim),조부경(Bu Kyung Jo),김민지(Min Ji Kim),강민주(Min Joo Kang),박기현(Ki-hyoun Park),구예은(Ye eun Koo),김병성(Byeongseong Kim),정의길(Eui-Gil Jung),한만득(Man-Deuk Han) 한국생명과학회 2015 생명과학회지 Vol.25 No.4

호두껍질을 둘러싸고 있는 과피는 식품으로서 효율성이 떨어지지만, 생리활성물질로서 항산화 및 항균효과의 가치를 지니고 있다. 본 연구에서는 호두(Juglans regia L.)의 열매과피를 5종류 용매(methanol, n-hexane, ethylacetate, n-butanol)로 추출하여 6종 추출물(MeOH Ex, Hexane Ex, CHCl3 Ex, EtOAc Ex, BuOH Ex, Water Ex)을 얻어, 총 폴리페놀과 총 플라보노이드함량을 특정하고, 항균 및 항산화 활성을 시험하였다. 각 추출물의 항균 활성은 병원균 8종에 대해 최소억제농도를 한천희석법으로 측정하였고, 항산화 활성은 DPPH (1,1-diphenyl-2-picrylhydrazyl)와 ABTS (2,2"-azino-bis-(3-ethylbenzothiazoline-6-sulfonate)) 라디칼 소거능을 측정하여 EC50 값으로 평가하고, 총 플라보노이드 및 페놀함량과 상관관계를 분석하였다. 호두추출물 가운데 항균활성이 가장 우수한 것은 에틸아세테이트 추출물(EtOAc Ex)로서 Staphylococcus aureus SG511에 대해 3.2 mg/ml의 MIC을 나타내었다. 또한, EtOAc Ex는 DPPH 및 ABTS 라디칼 소거 활성 EC50 값이 13.43 μg/ml과 41.83 μg/ml로 각각 나타내어 가장 우수한 항산화 활성을 보였다. 이러한 EtOAc Ex의 총 플라보노이드 및 페놀 함량은 각각 2.48 mg QE/g 및 223.25 mg GAE/g였다. 특히 총 페놀함량과 항균활성 및 항산화 활성은 양의 상관관계가 있어 추출액 속에 페놀함량이 높으면 생리활성도 높은 것이 확인되었다. 결론적으로 호두의 과피는 우수한 천연 항균 및 항산화 물질이 포함된 소재로서 이용 가능성을 제시한다. Several studies suggest that regular consumption of walnuts may have beneficial effects against oxidative stress-mediated disease such as cancer. The present study reports the total phenolic and flavonoid contents, together with the antioxidant and antibacterial activities of several solvent extracts (methanol, n-hexane, ethyl acetate, n-butanol, and water) obtained from walnut (Juglans regia L.) green husk. MIC (minimal inhibitory concentration) values of the walnut extracts for 8 human pathogenic bacteria strain were determined using agar dilution method. Antioxidant activity of extracts were assessed using DPPH (1,1-diphenyl-2-picrylhydrazyl) and ABTS (2,2"-azino-bis-(3-ethylbenzothiazoline-6-sulphonic acid)) assays, EC50 of DPPH and ABTS scavenging activities, and determination of total phenolic and flavonoid content and its correlation with DPPH and ABTS scavenging capacities. Among the six extracts, ethyl acetate extract (EtOAc Ex) showed the highest antimicrobial activity at 3.2 mg/ml of MICs against Staphylococcus aureus SG511. Total flavonoids and polyphenol contents of EtOAc Ex were 42.48 mg of quercetin equivalents (QE)/g and 223.25 mg of gallic acid equivalents (GAE)/g respectively. The highest antioxidative potential was shown by the sample extracted with EtOAc Ex (EC50=13.43 μg/ml for DPPH and EC50=41.83 μg/ml for ABTS radical scavenging activity assay). These results showed that J. regia green husk extracts can be used as an easily accessible source of natural antibacterial agents and natural antioxidants.