• 대한본초학회지
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• 제38권2호
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• pp.17-26
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• 2023

Objectives : A persistent inflammatory response can cause diseases such as fibrosis, cancer, and allergies. This study aimed to investigate the anti-inflammatory activity of Curcumae longae Rhizoma and Cinnamomi Ramulus Mixture (CCM) in lipopolysaccharide (LPS)-stimulated RAW264.7 cells. Methods : The total polyphenol and flavonoid contents of CCM were confirmed through an in vitro experiment. Also, radical scavenging activities of 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), and Hydroxyl were confirmed. Moreover, ferric reducing antioxidant power (FRAP) activity were confirmed. After, CCM (50, 100, and 200 ㎍/mL) were applied to 0.1 ㎍/mL LPS-stimulated RAW264.7 cells. The levels of nitric oxide (NO) and pro-inflammatory cytokines in the supernatant fraction were determined. Also, the expressions of mitogen-activated protein kinase (MAPK) and nuclear factor-κB (NF-κB) pathways were detected using Western blot. Results : As a result of in vitro experiments, there was an excellent antioxidant activity in CCM-treated cells. In addition, in RAW264.7 cells stimulated with LPS, the increased NO level was inhibited in a concentration-dependent manner by the treatment of CCM. In addition, inflammatory cytokines production were significantly inhibited in a concentration-dependent manner in CCM-treated group. CCM treatment significantly decreased the protein expressions of MAPKs. Moreover, the expressions of NF-κBp65 and cyclooxygenase-2 (COX-2) were significantly decreased when 200 mg/kg of CCM was applied, and phospho-inhibitor of nuclear factor kappa B-α (p-IκBα) and inducible nitric oxide synthase (iNOS) were significantly decreased at all concentrations treated with CCM. Conclusion : Our findings show that CCM exhibited excellent antioxidant activity and exhibited superior anti-inflammatory effect through the MAPKs and NF-κB pathways in LPS-stimulated RAW 264.7 macrophages.

• 대한본초학회지
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• 제38권5호
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• pp.49-60
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• 2023

Objectives : In this study, it was investigated the anti-inflammatory, anticancer, and antioxidant effects of Jayangdaebo-tang (JDT) consisting of twelve herbs before and after fermentation. Methods : JDT extract was fermented using the Lactoplantibacillus plantanum (JDT-L), Bacillus subtilis (JDT-B), and L. plantanum plus B. subtilis (JDT-L+B). The effects of each extract were measured in LPS-stimulated RAW264.7 cells, MCF-7 breast cancer and A549 lung cancer cells, and H₂O₂-stimulated HepG2 cells. Results : The extracts of JDT-L, JDT-B and JDT-L+B at 1 ㎎/㎖ decreased significantly the levels of nitric oxide (NO) in LPS-treated RAW264.7 cells and also inhibited the expression of iNOS and COX-2, and the phosphorylation of ERK and NF-κB. The JDT-L+B extract decreased significantly the expression of apoptotic proteins, Bax, cleaved caspase-3, and PARP in MCF-7 and A549 cancer cells. The JDT-L, JDT-B and JDT-L+B extracts increased significantly the cell viability in H₂O₂-stimulated HepG2 cells and the JDT-L+B extract decreased significantly the expression of SOD, catalase, HO-1, and NRF-2. Among fermented JDT extracts, JDT-L+B was the best effective on response of macrophage inflammation, cancer cell apoptosis, and liver cell damage. Conclusions : Our results were suggested that the fermentation can be used as a useful way to enhance the biological activity of JDT.

• 한국식품과학회지
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• 제47권1호
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• pp.119-125
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• 2015

본 연구에서는 들깨 유래 기능성 물질인 rosmarinic acid (RA)와 luteolin이 RAW264.7 세포에서 항염증작용에 대한 상승 효과가 있는지 알아보고자 하였다. 그 결과 RAW264.7 세포에 RA($50{\mu}M$)와 luteolin ($1{\mu}M$)을 동시에 처리하였을 경우 염증 매개인자인 NO, iNOS, $PGE_2$, COX-2의 생성을 RA ($100{\mu}M$)와 luteolin ($2{\mu}M$)을 각각 처리하였을 때 보다 더 뛰어나게 억제하였다. 또한 RA ($50{\mu}M$)와 luteolin ($1{\mu}M$)을 동시에 처리하였을 경우 TNF-${\alpha}$, IL-6, IL-$1{\beta}$ 같은 염증성 사이토카인의 생성량을 RA ($100{\mu}M$)와 luteolin ($2{\mu}M$)을 각각 처리하였을 때 보다 더 뛰어나게 억제하는 것을 확인하였다. 그리고 RA ($50{\mu}M$)와 luteolin ($1{\mu}M$)을 동시에 처리하였을 경우 RA ($100{\mu}M$)와 luteolin ($2{\mu}M$)을 각각 처리하였을 때 보다 NF-${\kappa}B$의 subunit인 p65의 translocation과 $I{\kappa}B$-${\alpha}$의 degradation을 더 뛰어나게 억제하는 것을 볼 수 있어 두 화합물 간의 상승작용이 뚜렷함을 확인 할 수 있었고, RA와 luteolin 두 화합물을 동시에 처리할 경우 염증관련 질환 치료에 유용하게 활용될 수 있을 것으로 판단된다.

• Asian-Australasian Journal of Animal Sciences
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• 제33권7호
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• pp.1167-1179
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• 2020

Objective: This study was conducted to fathom the underlying mechanisms of nutrition intervention and redox sensitive transcription factors regulated by Antrodia cinnamomea fermented product (FAC) dietary supplementation in broiler chickens. Methods: Four hundreds d-old broilers (41±0.5 g/bird) assigned to 5 groups were examined after consuming control diet, or control diet replaced with 5% wheat bran (WB), 10% WB, 5% FAC, and 10% FAC. Liver mRNA expression of antioxidant, inflammatory and lipid metabolism pathways were analyzed. Prostaglandin E2 (PGE₂) concentration in each group were tested in the chicken peripheral blood mononuclear cells (cPBMCs) of 35-d old broilers to represent the stress level of the chickens. Furthermore, these cells were stimulated with 2,2'-Azobis(2-amidinopropane) dihydrochloride (AAPH) and lipopolysaccharide (LPS) to evaluate the cell stress tolerance by measuring cell viability and oxidative species. Results: Heme oxygenase-1, glutathione S-transferase, glutamate-cysteine ligase, catalytic subunit, and superoxide dismutase, and nuclear factor (erythroid-derived 2)-like 2 (Nrf2) that regulates the above antioxidant genes were all up-regulated significantly in FAC groups. Reactive oxygen species modulator protein 1 and NADPH oxygenase 1 were both rather down-regulated in 10% FAC group as comparison with two WB groups. Despite expressing higher level than control group, birds receiving diet containing FAC had significantly lower expression level in nuclear factor-kappa B (NF-κB) and other genes (inducible nitric oxide synthase, tumor necrosis factor-α, interleukin-1β, nucleotide-binding domain, leucine-richcontaining family, pyrin domain-containing-3, and cyclooxygenase 2) involving in inflammatory pathways. Additionally, except for 3-hydroxy-3-methyl-glutaryl-coenzyme A reductase that showed relatively higher in both groups, the WB, lipoprotein lipase, Acetyl-CoA carboxylase, fatty acid synthase, fatty acid binding protein, fatty acid desaturase 2 and peroxisome proliferator-activated receptor alpha genes were expressed at higher levels in 10% FAC group. In support of above results, promoted Nrf2 and inhibited NF-κB nuclear translocation in chicken liver were found in FAC containing groups. H₂O₂ and NO levels induced by LPS and AAPH in cPBMCs were compromised in FAC containing diet. In 35-d-old birds, PGE₂ production in cPBMCs was also suppressed by the FAC diet. Conclusion: FAC may promote Nrf2 antioxidant pathway and positively regulate lipid metabolism, both are potential inhibitor of NF-κB inflammatory pathway.

• 아시안잔디학회지
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• 제25권1호
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• pp.89-93
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• 2011

본 연구는 질산희토비료의 시비에 따른 크리핑 벤트그래스의 생육에 미치는 영향을 평가하기 위해 수행되었다. 처리구는 비료의 시비량에 따라 무처리구(NF), 복합비료 만을 시비한 대조구(CF), 질산희토비료의 처리량에 따라 $0.3{\cdot}g{\cdot}m^{-2}$, 0.5 $g{\cdot}m^{-2}$, 1.0 $g{\cdot}m^{-2}$을 각각 처리한 처리구1(RE-1), 처리구2(RE-2), 처리구3(RE-3)이었고, 질산희토비료만을 처리한 처리구4(RE-4)였고, 각 처리구는 난괴법, 3반복으로 수행하였고, 엽색지수 및 엽록소지수와 같은 잔디품질, 잔디밀도, 건물중 및 잔디양분함유량을 조사하였다. 시험 전후 토양분석결과, 질산희토비료에 따른 토양화학성의 변화는 나타나지 않아 질산희토비료의 시비가 골프코스의 토양화학성의 변화에 영향을 미치지 않음을 확인하였다. 엽색지수와 엽록소지수를 조사한 결과, 질산희토비료의 시비 에 따라 비슷하거나 약간 증가하였고, RE-2의 처리가 가장 좋은 잔디품질을 나타내었다. 잔디밀도 조사 결과, 질산희토비료의 시비에 단일시비에 의해 9%증가하였고, 관행시비에 첨가하여 시비할 경우 RE-2에서 22%증가하였으며, 잔디 건물중은 통계적 유의성을 나타내지 않았다. 잔디 성분 중 질소함량이 질산희토비료를 처리한 잔디에서 높게 나타났다. 이러한 결과들을 통해 질산희토비료는 크리핑벤트그래스에서 질소흡수가 촉진되고, 잔디밀도를 향상되어 잔디품질을 향상시키는 가능이 있는 것으로 평가되었다.

• Journal of Periodontal and Implant Science
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• 제33권2호
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• pp.277-288
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• 2003

It has been suggested that increased number and activity of phagocytes in periodontitis lesion results in a high degree of reactive oxygen species (ROS) such as superoxide anion, hydrogen peroxide, nitric oxide and peroxynitrite. There are few reports on the relationship between ROS and MMPs expressions in gingival fibroblast. We studied to elucidate whether and how ROS, especially nitric oxide affects the MMP expression. Human gingival fibroblasts and HTl080 cells (human fibrosarcoma sell line as reference) were grown in DMEM supplemented with 10 mM HEPES, 50 mg/L gentamicin, and 10% heat inactivated fetal bovine serum with addition of various reactive oxygen species (ROS). Culture media conditioned by cells were examined by gelatin zymography. HT1080 cells expressed proMMP-2 and proMMP-9, but human gingival fibroblasts (HGF) produced only proMMP-2. Hydrogen peroxide upregulated MMP-9 expression in HT1080 cells, whereas in human gingival fibroblast SNP treatment showed marked increase in MMP-2 level compared to other ROS. These results suggest that the effects of ROS on MMPs expressions are cell-type specific. RT-PCR for MMP-2 and TIMP-2 m-RNA were performed using total RNA from cultured cells under the influence various kinase inhibitors. In HT1080 cells, treatment with FPTI III (Ras processing inhibitor) and LY294002 (PI3-kinase inhibitor) resulted in inhibition of MMP-2 and MMP-9 expressions, suggesting that Ras/P13-kinase pathway is important for MMPs expression in HT1080 cells. In gingival fibroblasts, treatment with FPTI III and PDTC (NF-kB inhibitor) showed marked decrease in MMP-2 regardless of the of SNP , suggesting that Ras/NF-kB could be the key pathway for NO-induced MMP-2 expression in gingival fibroblasts. This study showed that ROS, especially nitric oxide, could be the critical mediator of periodontal disease progression through control of MMP-2 expression in gingival fibroblasts possibly via Ras/NF-kB pathway.

• 한국식품과학회지
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• 제54권3호
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• pp.288-296
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• 2022

본 연구에서는 우선 진피 열수 추출물의 in vitro 항산화능을 평가하기 위해 total polyphenol, total flavonoid 함량, DPPH 및 ABTS radical 소거능을 분석한 후, 150 mM/60% ethanol로 유발한 급성 위염 동물 실험을 진행하여 급성 위염 완화 효과를 검증하였다. 약물투여군의 혈청 내 ROS와 MPO 수준, 조직 내 MDA 수준의 유의성 있는 감소를 확인하였으며, western blot을 통해 NOX2와 p22^phox를 포함한 산화적 스트레스 관련 단백질을 억제하였고, PI3K/Akt/NF- κB 신호 전달 경로를 통한 염증성 단백질의 현저한 감소를 확인하였다. 따라서 이러한 결과는 진피 열수 추출물이 급성 위염에 대한 완화 효과를 나타냈으며, 위염 및 천연 치료제의 후보 소재로서 가능성이 있다고 판단된다. 또한, 향후 만성 위염, 위암과 같은 위장 질환에 관한 추가 연구에서 진피 열수 추출물의 활용 가능성을 시사한다.

• Nutrition Research and Practice
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• 제16권6호
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• pp.729-744
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• 2022

BACKGROUND/OBJECTIVES: 4-Hydroxy-2-nonenal (HNE) is a biomarker for oxidative stress to induce inflammation. Methionine is an essential sulfur-containing amino acid with antioxidative activity. On the other hand, the evidence on whether and how methionine can depress HNE-derived inflammation is lacking. In particular, the link between the regulation of the nuclear factor-κB (NF-κB) signaling pathway and methionine intake is unclear. This study examined the link between depression from HNE accumulation and the anti-inflammatory function of ⳑ-methionine in rats. MATERIALS/METHODS: Male Wistar rats (3-week-old, weighing 70-80 g) were administered different levels of ⳑ-methionine orally at 215.0, 268.8, 322.5, and 430.0 mg/kg body weight for two weeks. The control group was fed commercial pellets. The hepatic HNE contents and the protein expression and mRNA levels of the inflammatory mediators were measured. The interleukin-10 (IL-10) and glutathione S-transferase (GST) levels were also estimated. RESULTS: Compared to the control group, hepatic HNE levels were reduced significantly in all groups fed ⳑ-methionine, which were attributed to the stimulation of GST by ⳑ-methionine. With decreasing HNE levels, ⳑ-methionine inhibited the activation of NF-κB by up-regulating inhibitory κBα and depressing phosphoinositide 3 kinase/protein kinase B. The mRNA levels of the inflammatory mediators (cyclooxygenase-2, interleukin-1β, interleukin-6, inducible nitric oxide synthase, tumor necrotic factor alpha) were decreased significantly by ⳑ-methionine. In contrast, the protein expression of these inflammatory mediators was effectively down regulated by ⳑ-methionine. The anti-inflammatory action of ⳑ-methionine was also reflected by the up-regulation of IL-10. CONCLUSIONS: This study revealed a link between the inhibition of HNE accumulation and the depression of inflammation in growing rats, which was attributed to ⳑ-methionine availability. The anti-inflammatory mechanism exerted by ⳑ-methionine was to inhibit NF-κB activation and to up-regulate GST.

• Journal of Nutrition and Health
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• 제56권5호
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• pp.455-468
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• 2023

본 연구에서는 잠재적인 기능성 소재로의 개발 가능성을 검증하기 위하여 미선나무 잎 추출물의 항산화능과 항염증 효능을 확인하였다. 시료의 총 폴리페놀함량, DPPH 및 ABTS 라디칼 소거능, FRAP value를 측정하여 미선나무 잎 추출물의 항산화 활성을 확인하였으며 또한 RAW264.7 세포에 미선나무 잎 추출물 처리 후 HO-1의 발현이 증가하는 것을 통해 미선나무 잎 추출물의 항산화 기전을 확인하였다. 한편, LPS로 유도된 염증 상태의 RAW264.7 세포에서 미선나무 잎 추출물은 NF-κB 활성 억제를 통한 NO 생성 억제, iNOS, COX-2 발현 억제 및 염증성 사이토카인의 발현과 생성을 억제하는 것을 확인하였다. 이러한 본 연구 결과는 향후 미선나무 잎 추출물의 기능성 소재로의 개발을 위한 기초자료 마련에 그 의의가 있다.

• 아시안잔디학회지
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• 제23권1호
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• pp.111-122
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• 2009

본 연구는 완효성 질소 성분이 함유된 복합비료의 시비가 크리핑벤드그래스 생육과 토양 중 변화에 미치는 영향을 시험하고자 2007년 7월부터 12월까지 6개월간 수행하였다. 처리구는 비료를 처리하지 않은 무처리구(NF), 속효성질소복합비료(11-5-7)를 처리한 대조구(CF), MU 질소성분이 함유된 복합비료를 처리한 처리구 1(MU), IBDU 질소성분이 함유 된 복합비료를 처리한 처리구 2(IBDU 1)과 처리구3(IBDU 2) 등 5개 처리를 난괴댑으로 수행하였다. 시비 후 속효성질소성분과 완효성질소성분의 경과시간에 따른 토양 중 변화와 염색지수와 엽록소함량과 같은 잔디품질과 잔디 생육량, 잔디밀도 등 잔디 생육을 조사하였고, 그 결과를 요약하면 아래와 같다. 암모니아태질소와 질산태질소는 모든 처리구에서 시비 후 시간이 경과함에 따라 점차증가하는 경향을 보였고, 총질소는 점차 감소하였다. 질산태질소와 총질소는 속효성 비료처리구(CF)보다는 완효성 비료 처리구(MU, IBDU 1, IBDU 2)에서 높게 나타났으며, 시비 후 경과시간에 따른 토양 중 가용성질소의 변화는 시비초기에는 속효성 비료의 가용성 질소가 많으나 시비 후 30일 정도까지는 속효성 비료와 완효성 비료의 가용성 질소 비율은 비슷하고 30일 이후에는 완효성 비료 처리구에서 가용성 질소 비율이 증가하였다. 염색지수와 엽록소함량을 NF와 비교한 결과, 엽색지수는 CF, MU, IBDU 1 및 IBDU 2에서 각각 6.5%, 6.7%, 5.9%, 5.5% 증가하였고, 엽록소함량은 33.2%, 38.4%, 35.1%, 37.1% 증가하여 MU 처리구에서 높게 나타났다. 잔디생육량은 CF, MU, IBDU 1 및 IBBU 2가 NF보다 76.2%, 77.7%, 69.5%, 72.3% 각각 높게 나타났으나 대조구와 처리구간 예초물량은 비슷하였고, 잔디밀도는 무처리구보다 비료 처리구에서 증가하였고, 속효성 비료처리구(CF)보다 완효성 비료 처리구(MU, IBDU 1, IBDU 2)에서 잔디밀도가 증가했다. 본 결과들을 통해 완효성 질소 성분함유비료가 속효성 질소에 비해 토양 중 가용성 질소의 함량을 높여 질소비효기간을 연장시키고, 염색지수와 엽록소함량은 비슷하며 예초물량은 비슷하거나 감소시키지만 잔디밀도는 향상시켜 완효성질소 함유비료의 시비가 잔디생육에 효과적인 것으로 평가되었다.