• Journal of Periodontal and Implant Science
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• 제34권3호
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• pp.607-622
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• 2004

Recently epidemiologic studies have indicated that the patients with periodontitis may have increased risk of ischemic cardiovascular events, and have suggested the important roles of blood cytokines and acute reactant proteins in the systemic infection and inflammatory response. Periodontitis and coronary heart disease (CHD) may share the common risk factors and the genetic mechanism associated with interleukin(IL)-1A, B and RA genotype may be involved in the production of IL-1. This study was aimed to investigate the relationship between angiographically defined CHD and periodontitis as chronic Gram-negative bacterial infection and to determine whether the IL-1 gene polymorphism is associated in both diseases. Patients under the age of 60 who had undergone diagnostic coronary angiography were enrolled in this study. Subjects were classified as positive CHD (+CHD, n=37) with coronary artery stenosis more than 50% in at least one of major epicardial arteries, and negative CHD (-CHD, n=30) without significant stenosis. After recording the number of missing teeth, periodontal disease severity was measured by means of plaque index (PI), gingival index (GI), bleeding on probing (BOP), probing depth (PD), clinical attachment level (CAL), and radiographic bone loss around all remaining teeth. Gingival crevicular fluid (GCF) was collected from the 4 deepest periodontal pockets and assessed for cytokine ($IL-1{\beta}$, IL-6, IL-1ra, tumor necrosis $factor-{\alpha}$, and prostaglandin $E_2$). Additionally, blood CHD markers, lipid profile, and blood cytokines were analyzed. IL-1 gene cluster genotyping was performed by polymerase chain reaction and enzyme restriction using genomic DNA from buccal swab, and allele 2 frequencies of IL-1A(+4845), IL-1B(+3954), IL-B(-511), and IL-1RA(intron 2) were compared between groups. Even though there was no significant difference in the periodontal parameters between 2 groups, GCF level of $PGE_2$ was significantly higher in the +CHD group(p<0.05). Correlation analysis showed the positive relationship among PD, CAL and coronary artery stenosis(%) and blood $PGE_2$. There was also significant positive relationship between the periodontal parameters (PI, PD, CAL) and the blood CHD markers (leukocyte count, C-reactive protein, and lactic dehyrogenase). IL-1 gene genotyping showed that IL-1A(+3954) allele 2 frequency was significantly higher in the +CHD group compared with the -CHD group (15% vs. 3.3%, OR 5.118,p=0.043). These results suggested that periodontal inflammation is related to systemic blood cytokine and CHD markers, and contributes to cardiovascular disease via systemic inflammatory reaction. IL-1 gene polymorphism might have an influence on periodontal and coronary heart diseases in Korean patients.

• 한국임상수의학회지
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• 제28권2호
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• pp.204-210
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• 2011

국내 독일 셰퍼드로부터 에를리히아증과 라임병에 대한 항체가 조사를 위하여 효소면역측정법을 시용하였으며, 중합효소연쇄반응을 이용하여 진드기매개성 병원체를 조사하였다. 2005년 10월부터 2006년 9월까지 국내 5개 지방으로부터 291마리 독일 셰퍼드를 선정하였다. 개 에를리히아 항체 조사를 위한 ELISA (Snap$^{(R)}$ 3Dx$^{(R)}$ test kit, IDEXX) 검사결과 전국적으로 7.56%(22마리)의 에를리히아 양성 항체 보균개와 1.72%(5마리)의 보렐리아 양성 항체 보균개로 조사되었다. 이 중 2마리에서는 두 가지 병원체 모두에 대한 양성 항체가 검출되었다. 에를리히아에 대한 양성항체를 지방별로 보면 경기도는 1.28%(78마리 중 1마리), 강원도는 12.64%(87마리 중 11마리), 충청도는 9.76%(41마리 중 4마리), 경상도는 8.93%(56마리 중 5마리), 전라도는 3.45%(56마리 중 5마리)로 조사되었다. Ehrlichia chaffeensis 유전자를 검출한 결과 전국 평균이 3.09%(291 마리 중 9마리), 강원도는 2.41%(291 마리 중 7마리) 그리고 충청도는 0.69%(291마리중 2마리)로 조사되었다. 중합효소연쇄반응으로 증폭된 유전자(SNU-EC3과 SNU-EC5)에 대한 유전자염기서열분석결과 한국(Haemaphysalis longicornis 진드기)과 중국 그리고 Arkansas 분리주인 E. chaffeensis와 매우 유사하였다. 이 연구의 결과 국내에서 사육되는 독일셰퍼드에서 에를리히아증의 병원체와 라임병에 대한 항체가 검출된 것으로 볼 때 임상 진단에 있어서 진드기매개전염병에 대한 중요성을 인식해야 할 것으로 판단된다. 따라서 혈소판감소증이나 백혈구감소증이 있는 경우 그리고 봄부터 가을까지 야외활동을 한 경력이 있는 환자에 대한 감별진단 항목에 반드시 포함시켜야 할 것을 제안한다.

• 한국미생물·생명공학회지
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• 제33권4호
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• pp.281-287
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• 2005

Paenibacillus polymyxa 유래의 endoinulinase 유전자(inu, 2.733 kb, E.C 3.2.1.7)를 Saccharomyces cerevisiae에 발현시키기 위해 대장균과 효모의 shuttle vector (GALl promoter함유)에 subcloning하여 구축된 pYGENIU27 (8.6 kb) plasmid를 S. cerevisiae SEY2102에 형질전환하였다. Uracil이 결핍된 SD배지와 inulin이 포함된 배지에서 효모 형질전환체를 선별하였다. 효모 형질전환체의 배양에서 endoinulinase는 periplasmic space에 다량 존재함이 확인되었다. YPDG배지에서 재조합 endoinulinase는 총 활성 1.81unit/ml으로 생산되었다. Inulooligosaccharides (IOSs) 생산을 위한 효소의 최적 반응 조건으로 pH 8.0,반응온도 $45^{\circ}C$, 기질은 Jerusalem artichoke로 결정되었다. 효소활성은 pH 10.0에서도 안정적으로 유지되었으며, 최적 반응 조건 (ginulin당 36 unit효소 이용)에서 반응 10분 후부터 IOSs가 생산되기 시작하였다. 생성된 IOSs의 구성분으로 inulobiose (F2), inulotriose (F3)와 inulotetraose (F4)가 생성되었고, 이중에서 F3가 주성분이었다. 이상의 결과는 효모에서 생산된 재조합 endoinulinase를 이용하여 inulin으로부터 기능성 감미소재인 IOSs의 산업적 생산을 위한 기초결과로 활용될 것이다.

• 대한한의학회지
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• 제27권1호
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• pp.184-195
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• 2006

Objectives : Betula Platyphylla(BP) is a traditional analgesic, anti-fungal, anti-inflammatory herb used in Chinese 1medicine. However, no information is available to explain its action. In this study. we investigated the anti-inflammatory 1effects of BP to elutidate the molecular pharmacological activity in the ethanol extract of BP(BPE). Methods : We performed WTS assay in lipopolysaccharide (LPS) stimulated RAW264.7 macrophages with BPE. Nitrite was measured by Griess assay, prostaglandin E2 (PGE2) by enzyme-linked immunosorbent assay (ELISA) in LPS induced RAW264.7 macrophages with BPE. Inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX-2) were determined by Western blot. Activation of nuclear factor-kappaB (NF-kB) was measured by electrophoretic mobility shift assay (EMSA). Results : BPE significantly suppressed production of nitric oxide (NO) and PGE2 in LPS-induced RAW264.7 macrophages in a dose-dependent manner. The maximal inhibition rate of NO and PGE2 production by BPE was ca. 88.8% and 93% at the concentration of $100{\mu}g/ml$ (non-cytotoxic concentration), respectively. BPE also decreased iNOS protein and COX-2 protein in LPS-induced RAW264.7 macrophages. EMSA demonstrated that BPE inhibited the DNA binding activity of the NF-kB. Conclusions : These results suggest that BPE inhibits NF-${\kappa}B$-mediated gene expression and downregulates inflammatory mediator production in RAW264.7 macrophages.

• 한국미생물·생명공학회지
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• 제42권3호
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• pp.275-284
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• 2014

본 연구에서는 식물자원으로부터 생리활성을 보유한 새로운 기능성 소재를 찾고자 하였다. 이를 위해 수종의 중국 자생 식물을 분석하여 강한 항산화능을 보유한 3종(Malus hupehensis, Ophiorrhiza cantonensis, 그리고 Psychotria rubra)을 선별하고, 각 추출물의 항산화능과 항염증 생리활성을 분석하였다. 먼저 각 추출물의 항산화능을 DPPH radical 소거능을 통해 분석한 결과 모두 양성 대조군으로 사용한 ascorbic acid와 유사한 정도의 높은 활성을 보여 각 소재가 매우 강한 항산화능을 보유함을 확인하였다. 또한 RAW 264.7 세포주에서 $H_2O_2$ 및 LPS에 의해 유도된 ROS에 대한 각 추출물의 소거능을 분석한 결과에서도 농도의존적인 강한 소거능을 보였다. 뿐만 아니라 대표적인 항산화효소 중 하나로 항산화능 보유 천연물에 의해 발현이 유도되는 HO-1 및 그 전사 인자인 Nrf-2의 단백질 발현이 각 추출물의 처리에 의해 증가됨을 보였다. 한편 각 소재가 LPS에 의해 유도된 NO 생성에 미치는 영향을 분석한 결과 모두 농도의존적인 NO 생성 저해능을 보였으며 이는 NO 생성 단백질인 iNOS의 발현 저해에서 기인함을 확인하였다. 이와 같은 각 소재의 NO 생성 및 iNOS 발현 억제 효과는 염증 상위신호 전달계인 NF-${\kappa}B$ 및 AP-1의 조절을 통해 일어날 가능성을 보였다. 이러한 결과를 통해 중국 자생 식물 3종의 높은 항산화능과 항염증 활성을 처음으로 확인하였으며 향후 기능성 소재로서 유용하게 활용될 수 있을 것으로 판단된다.

• 치위생과학회지
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• 제19권2호
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• pp.133-140
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• 2019

Background: The light-emitting diode (LED) curing light used is presumed to be safe. However, the scientific basis for this is unclear, and the safety of LED curing light is still controversial. The purpose of this study was to investigate the effect of LED curing light irradiation according to the conditions applied for the polymerization of composite resins in dental clinic on the cell viability and inflammatory response in Raw264.7 macrophages and to confirm the stability of LED curing light. Methods: Cell viability and cell morphology of Raw264.7 macrophages treated with 100 ng/ml of lipopolysaccharide (LPS) or/and LED curing light with a wavelength of 440~490 nm for 20 seconds were confirmed by methylthiazolydiphenyl-tetrazolium bromide assay and microscopic observation. The production of nitric oxide (NO) and prostaglandin $E_2$ ($PGE_2$) was confirmed by NO assay and $PGE_2$ enzyme-linked immunosorbent assay kit. Expression of interleukin $(IL)-1{\beta}$ and tumor necrosis factor $(TNF)-{\alpha}$ in total RNA and protein was confirmed by reverse transcription polymerase chain reaction and Western blot analysis. Results: The LED curing light did not affect the viability and morphology of normal Raw264.7 cells but affected the cell viability and induced cytotoxicity in the inflammation-induced Raw264.7 cells by LPS. The irradiation of the LED curing light did not progress to the inflammatory state in the inflammation-induced Raw264.7 macrophage. However, LED curing light irradiation in normal Raw264.7 cells induced an increase in NO and $PGE_2$ production and mRNA and protein expression of $(IL)-1{\beta}$ and $(TNF)-{\alpha}$, indicating that it is possible to induce the inflammatory state. Conclusion: The irradiation of LED curing light in RAW264.7 macrophage may induce an excessive inflammatory reaction and damage oral tissues. Therefore, it is necessary to limit the long-term irradiation which is inappropriate when applying LED curing light in a dental clinic.

• 대한약침학회지
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• 제25권3호
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• pp.242-249
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• 2022

Objectives: The aim of this work is to evaluate the in vitro antioxidant, hypoglycemic, and antiobesity effects of Euphorbia resinifera extracts and investigate the phenolic constituents and the toxicity of these extracts. Methods: Phytochemical screening was performed to detect polyphenols and flavonoids. Antioxidant activity was evaluated by four methods (DPPH, ABTS, H₂O₂, and xanthine oxidase inhibition). The hypoglycemic effect was determined by the inhibition of α-amylase and α-glucosidase enzymes in vitro and via a starch tolerance study in normal rats. The antiobesity effect was estimated by in vitro inhibition of lipase. Results: Phytochemical screening revealed that the ethanolic extract was rich in polyphenols (99 ± 0.56 mg GEA/g extract) and tannins (55.22 ± 0.17 mg RE/g extract). Moreover, this extract showed higher antioxidant activity in different tests: the DPPH assay (IC₅₀ = 53.81 ± 1.83 ㎍/mL), ABTS assay (111.4 ± 2.64 mg TE/g extract), H₂O₂ (IC₅₀ = 98.15 ± 0.68 ㎍/mL), and xanthine oxidase (IC₅₀ = 10.26 ± 0.6 ㎍/mL). With respect to hypoglycemic effect, the aqueous and ethanolic extracts showed IC₅₀ values of 119.7 ± 2.15 ㎍/mL and 102 ± 3.63 ㎍/mL for α-amylase and 121.4 ± 1.88 and 56.6 ± 1.12 ㎍/mL for α-glucosidase, respectively, and the extracts lowered blood glucose levels in normal starch-loaded rats. Additionally, lipase inhibition was observed with aqueous (IC₅₀ = 25.3 ± 1.53 ㎍/mL) and ethanolic (IC₅₀ = 13.7 ± 3.03 ㎍/mL) extracts. Conclusion: These findings show the antioxidant, hypoglycemic, and hyperlipidemic effects of E. resinifera extracts, which should be investigated further to validate their medicinal uses and their pharmaceutical applications.

• 대한본초학회지
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• 제33권6호
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• pp.35-42
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• 2018

Objectives : The aim of this study was to investigate whether the extract of Coptidis Rhizoma inhibits inflammation in chronic cold stress (CCS)-exposed mice or not. Methods : Coptidis Rhizoma extract (CRE) was made by reflux with distilled water. Male ICR mice (7 weeks old) were divided randomly into 5 groups: (1) control, (2) CCS, (3) CCS+CRE 100 mg/kg, (4) CCS+CRE 300 mg/kg, (5) CCS+CRE 1,000 mg/kg groups. Mice were orally administered once a day for 14 days starting from 1 day before CCS. Group (2)-(5) were exposed to CCS conditions that maintained at $4^{\circ}C$ for 2 h once a day for 14 days. The levels of serum cortisol and hypothalamic prostaglandin E1 (PGE1) and PGE2 were measured by enzyme-linked immunosorbent assay kit. The expression levels of several pro-inflammatory factors like heat shock protein 70 (HSP70), c-fos, and nuclear factor kappa-light-chain-enhancer of activated B cells (NF-kB) were measured by western blot analysis in mouse hypothalamus. Results : Oral administration of CRE 1,000 mg/kg significantly suppressed the increase of serum cortisol levels in mice exposed to CCS. CCS-exposed mice had significantly increased the expression of HSP70, c-fos, and NF-kB in hypothalamus, while CRE treatment significantly attenuated the elevation of these pro-inflammatory factors. The ratio of PGE2/PGE1 was also higher in CCS-exposed mice than control group. CRE treatment significantly reduced the increase of PGE2/PGE1 ratio induced by CCS. Conclusion : These findings suggest that Coptidis Rhizoma may work as a potential agent to modulate inflammatory responses under the condition of cold adaptation formed by CCS.

• 미생물학회지
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• 제52권2호
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• pp.192-201
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• 2016

최근 북극해의 축치해(Chuckchi Sea)로부터 저온지질분해 효소활성을 보이는 Psychrobacter sp. ArcL13 균주가 분리되었다. 그러나 낮은 발현 양 때문에, 이 효소의 다양한 분야에서의 활용에 제약을 받아왔다. 따라서 유전자 재조합 기술을 이용하여, 이 효소를 대량생산하는 기술개발이 필요하였다. 재조합 지질분해효소를 만들기 위해서는 우선 해당 유전자의 동정이 필요하였기 때문에, Psychrobacter sp. ArcL13 균주로부터 PCR을 이용한 gene prospecting 방법으로 새로운 지질분해효소 유전자인 ArcL13-Lip을 분리하고 전체 염기 서열을 규명하였다. 염기 서열 분석결과 ArcL13-Lip은 일부 Psychrobacter 속 박테리아 유래의 지질분해효소들과 염기 서열의 유사성은 낮지만, 84-90%의 아미노산 서열 유사성을 보였다. ArcL13-Lip 전체 유전자를 대장균에서 발현시키고 전기영동으로 분석한 결과, 재조합 ArcL13-Lip은 약 35 kDa의 분자량을 보였으며 단백질 봉입체 형태로 발현되었다. Unfolding된 ArcL13-Lip을 다양한 첨가물이 포함된 완충용액에서 refolding 시킨 결과, glucose에 의해서 refolding 효율이 가장 크게 증가하였다. Refolding된 재조합 ArcL13-Lip은 다양한 p-nitrophenyl ester 중 p-nitrophenyl caprylate과 p-nitrophenyl decanoate에 대해 가장 높은 효소활성을 보였다. 온도에 따른 효소활성을 조사한 결과 ArcL13-Lip은 $40^{\circ}C$에서 최고의 활성을 나타내었고, $10^{\circ}C$와 $20^{\circ}C$에서 각각 최고 활성 대비 약 40%와 73%의 효소활성을 나타내었다. 이와 같이 ArcL13-Lip은 전형적인 저온활성 효소의 특징을 보여주었다.

• Journal of Periodontal and Implant Science
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• 제26권2호
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• pp.448-468
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• 1996

To maintain its functional integrity, bone is continuously remodelled by a process involving resorption by osteoeclasts and formation by osteoblasts, In order to respond to changes in the physical environment or to trauma with the relevant action, this process is strictly regulated by locally synthesized or systemic fators, Prostaglandin $E_2(PGE_2$) is perhaps one of the best studied factors, having been known to affect bone cell function for several decades.$PGE_2$ has both anabolic and catabolic activities. Excess of $PGE_2$ has been implicated in a number of pathological states associated with bone loss in a number of chronic inflammatory conditions such as periodontal disease and rheumatoid arthritis. $PGE_2$ and other arachidonic acid metabolites have been shown to be potent stimulators of osteoclastic bone resorption in organ culture. The anabolic effects of $PGE_2$ were first noticed when an increase in periosteal woven bone formation was seen after the infusion of $PGE_2$ into infants in order to prevent closure of the ductus arteriosus. The cellular basis for the catabolic actions of $PGE_2$ has been well characterized. $PGE_2$increases osteoclast recruitment in bone marrow cell cultures. Also $PGE_2$ has a direct action on osteoclast serving to inhibit activity and can also indirectly activate osteoclast via other cells in the vicinity, presumably osteoblast. The cellular mechanisms for the anabolic actions of $PGE_2$ are not nearly so well understood. The purpose of this paper was to study the effects of $PGE_2$ and dibutyl(DB)cAMP on osteoblastic clone MC3T3El cells and on the generation of osteoclasts from their precursor cells. The effect of $PGE_2$ and DBcAMP on the induction of alkaline phoaphatase(AlP) was investigated in osteoblastic clone MC3T3El cells cultured in medium containing 0.4% fetal bovine serum. $PGE_2$ and DBcAMP stimulated ALP activity and MTT assay in the cells in a dose-dependent manner at concentrations of lO-SOOng/ml. Cycloheximide, protein synthesis inhibitor, inhibited the stimulative effect of $PGE_2$ and DBcAMP on ALP activity in the cells. $PGE_2$also increased the intracellular cAMP content in a dose-dependent fashion with a maximal effect at 500ng/ml. The effect of $PGE_2$ on the generation of osteoclasts was investigated in a coculture system of mouse bone marrow cells with primary osteoblastic cells cultured in media containing 10% fetal bovine serum.After cultures, staining for tartrate-resistant acid phosphatase(TRAP)-marker enzyme of osteoclast was performed. The TRAP(+) multinucleated cells(MNCs), which have 3 or more nuclei, were counted. More TRAP(+) MNCs were formed in coculture system than in control group. $PGE_2(10^{-5}10^{-6}M)$ stimulated the formation of osteoclast cells from mouse bone marrow cells in culture. $PGE_2(10^{-6}M)$ stimulated the formation of osteoclast cells from mouse bone marrow cells in coculture of osteoblastic clone MC3T3E1 cells This results suggest that $PGE_2$ stimulates the differentiation of osteoblasts and generation of osteoclast, and are involved in bone formation, as well as in bone resorption.