• Journal of Periodontal and Implant Science
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• 제23권2호
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• pp.243-259
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• 1993

The bone resorbing activity of $PGE_2$ and elevated level of prostaglandins(PGs) and thromboxanes (TXs) in inflamed gingiva which are cyclooxygenase(C) metabolites have been well documented. Nonsteroidal anti-inflammatory drugs(NSAIDs) have been known to suppress gingival inflammation and bone resorption through the specific inhibitory action on the C pathway thereby decrease of various C metabolites. Recent studies provide unequivocal results that gingival tissue metabolizes arachidonic acid(AA) mainly through lipoxygenase(L) pathway. And the results of our previous experiments suggest that indomethacin may have inhibitory action on L as well as C. Thus we started this study to show the influences of several C inhibitors on the L activity at therapeutic and toxic dosage. Periodontal tissue samples were obtained from patients with advanced periodontitis and incubated with $^{14}C-AA(0.2{\mu}Ci)$ and various enzyme inhibitors. The tissue lipid extracts were separated by means of thin layer chromatography(TLC) and analyzed by means of autoradiography and TLC analyzer. Our results showed that aspirin inhibited C more selectively than L, however at higher concentration it also decreased HETEs production significantly. Indomethacin showed dose-dependent inhibition of L as well as C and all of the L metabolites were decreased to the same degree by high concentration of indomethacin. AA-861, which is an experimental tool of selective L inhibitor, showed inhibition of HETEs production but no effect on the production of $TXB_2$, PGs and $LTB_4$. Various propionic acid derivatives NSAIDs(ibuprofen, flurbiprofen, naproxen) showed the same patterns of effect on AA metabolism each other that was profound inhibition of PGs production, to the less degree HETEs and $TXB_2$ production, and of no effect on the $LTB_4$ production.

• Journal of Applied Biological Chemistry
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• 제60권3호
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• pp.191-198
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• 2017

잣나무 잎 추출물에 대해 염증인자인 hyaluronidase 저해효과를 측정한 결과 ethanol 추출물에서 물추출물에 비해 상대적으로 더 높은 HAase 저해효과를 보여주었으며, $50-200{\mu}g/mL$ phenolic 농도에서 13.2-29.5%의 저해효과를 나타내었다. 잣나무잎 추출물을 Raw 264.7 cell에 농도별로 처리하여 생성되는 NO량을 측정한 결과 LPS처리군은 LPS 무처리군에 비하여 3배에 가까운 NO발현을 나타내었으며, 농도 의존적으로 NO발현을 억제하였다. LPS에 의해서 생성된 $PGE_2$의 생성억제 효과를 측정한 결과 $25{\mu}g/mL$의 처리농도에서 ethanol 추출물이 26.2%의 저해율을 나타내었다. NO 생성 억제기작에 관한 iNOS 단백질의 발현량을 측정한 결과 $25{\mu}g/mL$의 농도에서 40%의 높은 억제효과를 나타내었으며, 농도 의존적으로 감소하는 경향을 관찰 할 수 있었다. LPS 처리시 또 다른 염증인자인 COX-2의 단백질 발현을 측정한 결과 $25{\mu}g/mL$의 처리 농도에서 64%의 저해효과를 나타내었다. 잣나무잎 추출물이 Raw 264.7 세포에서 LPS에 의해서 생성되는 $TNF-{\cdot}$, IL-6의 생성억제 효과를 측정한 결과 $25{\mu}g/mL$의 처리농도에서 각각 61.7, 62%의 저해 효과를 나타내었다. 따라서 잣나무잎 추출물은 염증억제를 위한 천연 기능성 소재로 활용이 가능할 것으로 판단되었다.

• 생명과학회지
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• 제26권11호
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• pp.1245-1252
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• 2016

진달래꽃으로부터 Sephadex LH-20 및 MCI gel CHP-20 column chromatography로 정제한 결과 항염증 활성을 가지는 myricetin을 분리, 동정하였다. Myricetin은 농도 의존적으로 NO 발현을 억제하였고, $50{\mu}M$ 농도에서 약 40%의 억제효과를 나타내었다. Myricetin의 iNOS와 COX-2의 발현억제력은, $25{\mu}M$ 농도에서 각각 20% 및 80%의 protein 발현 억제효과를 나타내었다. 또한 myricetin의 염증반응의 cytokine을 측정하여 $TNF-{\alpha}$, $IL-1{\beta}$, IL-6 및 $PGE_2$의 억제력을 살펴본 결과, 농도 의존적으로 발현억제 효과를 나타내었으며, $50{\mu}M$ 농도에서 각각 70%, 80%, 80% 및 95%의 발현 억제효과를 나타내었다. 따라서 진달래 꽃잎에서 분리한 myricetin은 LPS로 유도되어진 대식세포주인 Raw 264.7 세포에서 염증반응의 억제효과를 기대할 수 있었다.

• 생약학회지
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• 제38권4호
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• pp.339-348
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• 2007

This study were designed to evaluate the anti-inflammatory effects of $genistein-4'-O-{\alpha}-L-rhamnopyranosyl-(1-2)-{\beta}-D-glucopyranoside$ (GRG) isolated from Sophora japonica (Leguminosae) on the lipopolysaccharide (LPS)-induced nitric oxide (NO) and prostaglandin ($PGE_2$) production by RAW 264.7 cell line. GRG significantly inhibited the LPS-induced NO and $PGE_2$ production. Consistent with these observations, GRG reduced the LPS-induced expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) at the protein and mRNA levels in a concentration-dependent manner. In addition, the release and the mRNA expression levels of tumor necrosis $factor-{\alpha}\;(TNF-{\alpha})$ and interleukin-6 (IL-6) were also reduced by GRG. Moreover, GRG attenuated the LPS-induced activation of nuclear factor-kappa B ($NF-{\kappa}B$), a transcription factor necessary for pro-inflammatory mediators, iNOS, COX-2, $TNF-{\alpha}$ and IL-6 expression. These results suggest that the down regulation of iNOS, COX-2, $TNF-{\alpha}$, and IL-6 expression by GRG are achieved by the downregulation of $NF-{\kappa}B$ activity, and that is also responsible for its anti-inflammatory effects.

• Korean Journal of Acupuncture
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• 제21권2호
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• pp.125-137
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• 2004

Objectives : Recently, Herbal-acupuncture therapeutics has been used for the treatment of inflammatory diseases such as rheumatoid arthritis. Especially, we have been interested in chemical mediators concerned with inflammation such as prostaglandin, cytokine, nitrous oxide. The purpose of this study is investigated that the effect of Chelidonii Herbal-acupuncture solution in lipopolysaccharide-stimulated RAW 264.7 macrophages, performed several expeimental items : those are prostaglandin $E_2$, nitric oxide and cyclooxygenase-2. Methods : The cytotoxicity of Chelidonii Herbal-acupuncture solution in RAW 264.7 macrophages were measured by MTT-based cytotoxicity assay. In order to observe cyclooxygenase-2 mRNA expression in lipopolysaccharide-stimulated RAW 264.7 macrophages, RT-PCR was used. Prostaglandin $E_2$ formation and nitric oxide production was measured by competitive enzyme immunoassay and Griess assay. Results : 1.The MTT assay demonstrated that cytotoxic effect of Chelidonii Herbal-acupuncture solution in RAW 264.7 macrophages were not appeared before concentration of 1mg/ml. 2.Chelidonii Herbal-acupuncture solution inhibited cyclooxygenase-2 mRNA expression in lipopolysaccharide-stimulated RAW 264.7 macrophages. 3. Chelidonii Herbal-acupuncture solution inhibited nitric oxide production in lipopolysaccharide-stimulated RAW 264.7 macrophages. 4. Chelidonii Herbal-acupuncture solution inhibited prostaglandin $E_2$ formation in lipopolysaccharide-stimulated RAW 264.7 macrophages. Conclusions : On the basis of these results, It was shown that Chelidonii Herbal-acupuncture solution is significantly able to inhibit the production of $PGE_2$ and NO, as well as COX-2 mRNA expression. Our results may provide new mechanism by which Chelidonii Herbal-acupuncture solution accounts for its beneficial effect on accelerating wound healing and anti-inflammation.

• 대한한방부인과학회지
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• 제34권3호
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• pp.29-48
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• 2021

Objectives: Banchong-san (BCS) is a herbal formula composed of 13 korean medicinal herbs and is traditionally used to treat inflammatory diseases and pain. The object of this study was to research the antioxidant, anti-inflammatory, antipruritic and antimicrobial effects of the BCS in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. Methods: In this experiment, effects of BCS on the following four were measured as follows: (1) Anti-oxidative effects were evaluated by 1,1-diphenyl-2-picryl-hydrazyl (DPPH) Radical scavenging activity, 2,2-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) Radical scavenging activity. (2) Anti-inflammatory effects were evaluated by the production amount of Reactive oxygen species (ROS), Nitric oxide (NO), Interleukin-1β (IL-1β), Interleukin 6 (IL-6), tumor necrosis factor-α (TNF-α), Prostaglandin E2 (PGE₂), inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2)(the previous two are "mRNA"), extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), p38 mitogen-activated protein kinases (p38), inhibitor of nuclear factor kappa B (IκBα), nuclear factor kappa B (NF-κB) (the previous five are "Protein") in LPS-Stimulated RAW 264.7 cells. (3)Antipruritic effects were evaluated by the production amount of histamine, Leukotriene B4 (LTB4), LeukotrieneC4 (LTC4) Levels in phorbol 12-myristate 13-acetate(PMA)/ionomycin-stimulated MC/9 mast cell. (4) Anti-microbial effects were evaluated by the growth suppression of Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa and Aspergillus niger. Results: The following results were obtained through each measurement: (1) DPPH Radical Scavenging Activity, ABTS Radical Scavenging Activity evoked a significant concentration-dependent increase. (2) ROS, NO, IL-1β, IL-6, TNF-α, PGE₂ production amount, iNOS, COX-2 mRNA expression were significantly reduced in the BCS extraction group compared with the control group and significantly decreased the amount of ERK, JNK, p38, NF-κB Protein expression. The amount of IκB-α Protein Expression have increased significantly. (3) The amounts of histamine, LTB4, LTC4 were significantly decreased. (4) The antibacterial efficacy, BCS inhibited the growth of Escherichia coli, Pseudomonas aeruginosa at concentrations of 5 ㎍/ml, but did not suppress the growth of staphylococcus aureus and aspergillus niger. Conclusions: The experimental results show that BCS has anti-oxidant, anti-inflammatory, antipruritic and antimicrobial properties.

• Restorative Dentistry and Endodontics
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• 제17권1호
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• pp.83-94
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• 1992

This study was executed to measure the biosynthesis of arachidonic acid metabolic products in chronic periapical lesions, to compare the products among periapical granuloma, periapical cyst and chronic periapical abscess, and to understand the pathogensis of chronic periapical lesions. Tissues from 33 chronic periapical lesions of human teeth were enucleated during endodontic surgery. large part of each tissue was contained in liquid nitrogen immediately and the other was examined histologically. In histologically diagnosed 8 cases of periapical granuloma, 9 cases of periapical cyst and 8 cases of chronic periapical abscess. the tissues were homogenatecl and incubated with $_{14}C$-arachidonic acid. Lipid solvent extracts were separated by thin layer chromatography to be analyzed by autoradiography and TLC analyzer. 1. $TXB_2$, 6-keto-$PGF_1{\alpha}$ and $PGE_2$, $LTB_4$, HETEs, and unidentified product which are metabolic products of arachidonic acid were measured in the tissues of chronic peripaical lesions. 2. In all of periapical granuloma, cyst and abscess, the conversion rate of HETEs among all products was the highest(P<0.05), and the percentage of HETEs in total converted products was also the highest(P<0.05). 3. The concentration of each arachidonic acid product was higher in chronic periapical absecss than in periapical granuloma and cyst(P<0.05). The concentration of $TXB_2$ and HETEs in periapical cyst were hight than in periapical granuloma. 4. The relative amounts of total products from lipoxygenase pathway to those from cyclo-oxygenase pathway were about 7 fold in chronic periapical lesions. There was no difference among periapical granuloma, cyst and abscess(P<0.05). The total amount of products from each pathway were higher in chronic periapical abscess than in periapical cyst and granuloma.

• Journal of Periodontal and Implant Science
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• 제37권1호
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• pp.91-102
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• 2007

The long term success of periodontal treatment is dependent upon the effectiveness of the main-tenance care program after active treatment. The purpose of this study was to evaluate whether nutraceutical containing PRF-K2 as natural product from plant and seaweed has beneficial effects on clinical parameters, gingival crevicular fluid (GCF) volume and GCF cytokine levels during main- tenance phase after periodontal treatment. Among the generally healthy and non-smoking. moderate to severe chronic periodontitis patients during maintenance phase in Department of Periodontics, Chonnam National University Hospital, twenty eight patients took nutraceutical containing PRF-K2 (Oscotec Inc. Cheonan, Korea) for 3 months as experimental group and sixteen patients received only maintenance care as control group. Clinical examination and GCF collection were performed at baseline, 1, 2 and 3 months of experiment. Total amounts and concentrations of GCF IL-1{\beta}, IL-1ra and $PGE_2$ were evaluated using ELISA kit. In probing pocket depth, experimental group showed the tendency of more reduction than control group after 3 months of experiment. Sulcus bleeding index (SBI) and GCF volume were significantly decreased in experimental group(p<0.05), whereas they were increased in control group. GCF IL-1{\beta} level tended to decrease in both experimental and control group and IL-1ra concentration tended to increase in experimental group and to decrease in control group. IL-1ra/IL-1{\beta} ratio tended to increase in experimental group and to decrease in control group during experimental period. GCF $PGE_2$ amount did not show any change in experimental group and tended to increase in control group. These results suggest that nutraceutical supplement which contain PRF-K2 could improve perio-dontal condition during maintenance phase after periodontal therapy.

• Journal of Dairy Science and Biotechnology
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• 제38권2호
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• pp.112-120
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• 2020

본 연구는 Enterococcus faecalis EF-2001 사균체를 발효유에 첨가하였을 시 나타나는 생리활성을 검증하고자 실시하였다. 발효유는 같은 양의 starter만 넣은 일반발효유 NFM, E. faecalis EF-2001 사균체를 100 ㎍/mL의 농도로 첨가한 EFM1, E. faecalis EF-2001 사균체를 500 ㎍/mL의 농도로 넣은 EFM2를 제조하였다. E. faecalis EF-2001 사균체를 발효유 첨가하였을 때 나타나는 항염증 효과를 검증하기 위해 mouse 유래 대식세포인 RAW 264.7 cell을 이용하였다. RAW 264.7 cell을 이용한 세포독성 실험의 결과, 사균의 농도가 높아질수록 세포 생존율이 유의적으로 감소하였으며(p<0.05), 80% 이상의 세포 생존율을 가지는 농도인 5, 10, 15 ㎍/mL에서 실험을 진행하였다. Nitric oxide의 생성 억제능 측정결과는 LPS 처리군에 비해 발효유에서 NO 생성을 저해하는 경향을 나타냈다. 또한, 일반발효유보다 사균을 첨가한 발효유에서 NO 생성을 더욱 저해하는 결과를 나타내었다. Prostaglandin E₂의 억제율 측정결과도 15 ㎍/mL의 농도에서 PGE₂ 분비량을 감소시킴을 확인하였으며, 일반 발효유보다 사균이 첨가된 발효유에서 더욱 억제됨을 확인하였다. 따라서, 본 연구를 통해 E. faecalis EF-2001 사균체가 발효유에 첨가되어도 염증 매개물질인 NO 및 PGE₂의 생성을 감소시킴으로써 추후 사균체가 첨가된 발효유 제조 및 연구에 대한 기초 연구 결과로 활용될 수 있을 것으로 사료된다.

• 생명과학회지
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• 제32권7호
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• pp.491-500
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• 2022

HK표고버섯균사체(HK shiitake mushroom mycelium, HKSMM)는 간 건강 개별 인정 건강기능식품이다. LPS로 활성화된 RAW 264.7 세포에서 HKSMM50 (HKSMM의 50% ethanol 수용액 추출물)의 항염증효과를 연구하였다. AHCC는 positive control로 사용하였다. LPS로 활성화된 RAW 264.7 세포에 HKSMM50 및 AHCC를 처리(0, 20, 100, 500 ㎍/ml)하고 24시간 배양하여 배양물의 염증 관련 인자는 ELISA kits로, 세포에 함유된 iNOS와 COX-2 protein 발현은 Western blotting으로 측정하였다. HKSMM50는 LPS 처리에 비해 농도 의존적으로 NF-κB 함량을 낮추었고, iNOS와 COX-2 protein 발현을 억제하여 NO와 PGE₂ 함량을 낮추었다. 더불어 HKSMM50는 LPS 처리에 비해 IL-1β, TNF-α, IL-4 및 IL-6의 함량을 낮추었으나 SOD와 CAT의 활성은 증가시켰다. AHCC도 HKSSM50 처리와 비슷한 효과를 나타내었다. 이 결과는 HKSMM50이 LPS로 활성화된 RAW 264.7 세포에서 NF-κB 신호전달을 억제하여 항염증효과를 나타내었으며, HKSMM은 면역기능증진에 도움을 줄 수 있는 건강기능식품원료로 사용할 수 있을 것이다.