• 한국식품과학회지
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• 제38권5호
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• pp.699-706
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• 2006

최근 약용식물로부터 특정성분을 추출하여 천연물이 가지는 2차 대사산물인 생리활성 물질에 대한 관심이 증대되고 있는데, 생리활성을 나타내는 물질 중 식품의 기능 강화를 위해 주로 활용되고 있는 것은 항산화 및 항균활성에 관련된 물질들이다. 이러한 식품강화 물질을 발굴하고자 해안가에 자생하며 유독성 식물로 알려진 암대극(E. jolkini)의 전초를 대상으로 ethanol 추출 및 용매분획하여 총 폴리페놀 함량을 분석하고 그에 따른 항산화활성 및 식품 유해 미생물에 대한 항균활성을 비교 측정하였다. 그 결과 암대극 (E. jolkini)은 다른 대극속 식물과 마찬가지로 페놀성 화합물을 다수 함유하고 있음을 확인할 수 있었으며, 총 폴리페놀 함량이 높게 나타난 ethylacetate 층에서 DPPH radical 소거활성, xanthine oxidase 저해활성, superoxide 소거활성 및 nitric oxide 소거활성의 $IC_{50}$ 값이 각각 $8.38\;{\mu}g/mL$, $466.01\;{\mu}g/mL$, $11.39\;{\mu}g/mL$, $332.11\;{\mu}g/mL$로 가장 우수한 항산화 활성을 보여 주었다. 항균활성은 paper disc method, 미생물 생장곡선 그리고 최대억제농도(MIC)을 측정 비교하였다. Paper disc method에서는 조추출물인 80% ethanol 및 ethylacetate와 butanol 분획물에서 농도가 증가할수록 항균력을 나타내는 inhibition zone 크기도 같이 증가하였으며, 특히 ethylacetate 분획물에서 가장 높은 항균력을 보여 주었다. 최대억제농도는 모든 균주에서 80% ethanol 추출물 및 ethylactate와 butanol 분획물에서 100 ppm 농도로 처리시 최소 생육저해 효과를 보였으며, 또한 암대극 추출물이 식품유해 미생물인 B. cereus, L. monocytogenes, E. coli 등 3 종의 균주를 대상으로 생육특성에 미치는 영향을 측정한 결과, ethylacetate 분획물이 가장 효율적인 항균활성을 보여 주었다.

• 한국식품영양과학회지
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• 제43권10호
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• pp.1527-1534
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• 2014

Lactobacillus rhamnosus로 발효시킨 흑마늘 발효물의 항염증 효능을 검증하기 위해 LPS로 염증 유도된 RAW 264.7 cells를 이용하여 관련 인자들을 분석하였다. 100, 200, 400 및 $800{\mu}g/mL$ 농도에서 세포독성은 유발되지 않았으며, 오히려 농도 의존적으로 세포 생존율은 증가하였다. LPS에 의해 염증 유도된 RAW 264.7 cells에서 흑마늘 발효물은 농도 의존적으로 NO와 $PGE_2$의 생성 감소와 염증성 사이토카인인 TNF-${\alpha}$, IL-$1{\beta}$ 및 IL-6의 단백질 생성을 감소시켰다. 또한 iNOS, COX-2, NF-${\kappa}B$ 및 $I{\kappa}B$ 단백질의 발현을 감소시키고 HO-1의 단백질의 발현을 증가시켰다. 이상의 연구 결과를 통해 흑마늘 발효물은 염증에 의한 NF-${\kappa}B$의 활성과 TNF-${\alpha}$, IL-$1{\beta}$와 IL-6의 생성을 억제시키고, iNOS 및 COX-2의 발현을 억제시키는 메커니즘을 통해 염증성 질환의 예방 및 개선 효능을 나타내는 것으로 판단된다.

• BMB Reports
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• 제55권6호
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• pp.275-280
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• 2022

The treatment of atopic dermatitis (AD) is challenging due to its complex etiology. From epidermal disruption to chronic inflammation, various cells and inflammatory pathways contribute to the progression of AD. As with immunosuppressants, general inhibition of inflammatory pathways can be effective, but this approach is not suitable for long-term treatment due to its side effects. This study aimed to identify a plant extract (PE) with anti-inflammatory effects on multiple cell types involved in AD development and provide relevant mechanistic evidence. Degranulation was measured in RBL-2H3 cells to screen 30 PEs native to South Korea. To investigate the anti-inflammatory effects of Parasenecio auriculatus var. matsumurana Nakai extract (PAE) in AD, production of cytokines and nitric oxide, activation status of FcεRI and TLR4 signaling, cell-cell junction, and cell viability were evaluated using qRT-PCR, western blotting, confocal microscopy, Griess system, and an MTT assay in RBL-2H3, HEK293, RAW264.7, and HaCaT cells. For in vivo experiments, a DNCBinduced AD mouse model was constructed, and hematoxylin and eosin, periodic acid-Schiff, toluidine blue, and F4/80-staining were performed. The chemical constituents of PAE were analyzed by HPLC-MS. By measuring the anti-degranulation effects of 30 PEs in RBL-2H3 cells, we found that Paeonia lactiflora Pall., PA, and Rehmannia glutinosa (Gaertn.) Libosch. ex Steud. show an inhibitory activity of more than 50%. Of these, PAE most dramatically and consistently suppressed cytokine expression, including IL-4, IL-9, IL-13, and TNF-α. PAE potently inhibited FcεRI signaling, which mechanistically supports its basophil-stabilizing effects, and PAE downregulated cytokines and NO production in macrophages via perturbation of toll-like receptor signaling. Moreover, PAE suppressed cytokine production in keratinocytes and upregulated the expression of tight junction molecules ZO-1 and occludin. In a DNCB-induced AD mouse model, the topical application of PAE significantly improved atopic index scores, immune cell infiltration, cytokine expression, abnormal activation of signaling molecules in FcεRI and TLR signaling, and damaged skin structure compared with dexamethasone. The anti-inflammatory effect of PAE was mainly due to integerrimine. Our findings suggest that PAE could potently inhibit multi-inflammatory cells involved in AD development, synergistically block the propagation of inflammatory responses, and thus alleviate AD symptoms.

• Journal of Ginseng Research
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• 제45권2호
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• pp.344-353
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• 2021

Background: Korean Red ginseng extract (KRGE) has beneficial effects on the cardiovascular system by improving endothelial cell function. However, its pharmacological effect on endothelial cell senescence has not been clearly elucidated. Therefore, we examined the effect and molecular mechanism of KRGE on the senescence of human umbilical vein endothelial cells (HUVECs). Methods: HUVECs were grown in normal or KRGE-supplemented medium. Furthermore, they were transfected with heme oxygenase-1 (HO-1) gene or treated with its inhibitor, a NF-κB inhibitor, and a miR-155-5p mimic or inhibitor. Senescence-associated characteristics of endothelial cells were determined by biochemical and immunohistochemical analyses. Results: Treatment of HUVECs with KRGE resulted in delayed onset and progression of senescence-associated characteristics, such as increased lysosomal acidic β-galactosidase and decreased telomerase activity, angiogenic dysfunction, and abnormal cell morphology. KRGE preserved the levels of anti-senescent factors, such as eNOS-derived NO, MnSOD, and cyclins D and A: however, it decreased the levels of senescence-promoting factors, such as ROS, activated NF-κB, endothelial cell inflammation, and p21 expression. The beneficial effects of KRGE were due to the induction of HO-1 and the inhibition of NF-κB-dependent biogenesis of miR-155-5p that led to the downregulation of eNOS. Moreover, treatment with inhibitors of HO-1, NF-κB, and miR-155-5p abolished the anti-senescence effects of KRGE. Conclusion: KRGE delayed or prevented HUVEC senescence through a signaling cascade involving the induction of HO-1, the inhibition of NF-κB-dependent miR-155-5p biogenesis, and the maintenance of the eNOS/NO axis activity, suggesting that it may protect against vascular diseases associated with endothelial senescence.

• 대한한방내과학회지
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• 제29권1호
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• pp.130-148
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• 2008

Objective : This study was designed to investigate the effect of the water extract of Gamisamgibopae-tang(GMSGBPT), an oriental herbal formulation, on the growth of NCI-H460 and A549 human non-small-cell lung cancer cell lines. Methods : Cytotoxicity and cell morphology were evaluated by MTT assay and inverted microscope, respectively. Apoptosis was detected using agarose gel electrophoresis and flow cytometer. The expression levels of mRNAs and proteins of target genes were determined by RT-PCR and western blot analyses, respectively Result and Conclusion : We found that exposure of A549 cells to GMSGBPT resulted in the growth inhibition in a dose-dependent manner as measured by MTT assay, but GMSGBPTdid not affect the growth of NCI-H460 cells. The anti-proliferative effect of GMSGBPT treatment in A549 cells was associated with morphological changes, formation of apoptotic bodies and DNA fragmentation, and flow cytometry analysis confirmed that GMSGBPT treatment increased the populations of apoptotic-sub G1 phase. Growth inhibition and apoptotic cell death by GMSGBPT were connected with a up-regulation of cyclin-dependent kinase inhibitor p21 (WAF1/CIP1) mRNA and protein in a tumor suppressor p53-independent fashion. However GMSGBPT treatment did not affect other growth regulation-related genes such as early growth response-1 (Egr-1), nonsteroidal anti-inflammatory drug (NSAID)-activated gene-1 (NAG-1), inducible nitric oxide synthase (iNOS), cyclooxygenases (COXs), telomere-regulatory factors in A549 orNCI-H460 cells. Taken together, these findings partially provide novel insights into the possible molecular mechanism of the anti-cancer activity of GMSGBPT.

• 한국자원식물학회지
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• 제25권6호
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• pp.682-692
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• 2012

Sea Buckthorn (Hippophae rhamnoides L.) has been used in traditional medicine for the treatment of cough, indigestion, circulatory problems and pain. The associated anti-inflammatory effect of this agent is achieved via the inhibition of Nf-${\kappa}B$ signaling, a property that has been demonstrated to effectively control the symptoms of various skin disorders, including atopic dermatitis. Accordingly, the purpose of this study was to assess the efficacy of Sea Buckthorn in reducing the production of lipopolysaccharide (LPS) activated nitric oxide (NO) by inhibiting the Nf-${\kappa}B$ pathway, as measured by the symptoms of atopic dermatitis (AD) occurring secondarily to inflammation and immune dysregulation. Our data demonstrate that Sea Buckthorn significantly decreased the LPS-induced production of NO (p<0.001). Atopic dermatitis was induced by repeated application of 2,4-dinitrochlorobenzene to the dorsal skin of mice. Topical application of 5% Sea Buckthorn extract improved the symptoms of AD, specifically reducing disease severity scores, scratching behaviors and epidermal thickness. When compared to the control group, animals treated with Sea Buckthorn exhibited increased serum IL-12 levels and decreased serum TNF-${\alpha}$, IL-4 and IL-5 levels. Such a modulation of biphasic T-helper (Th)1/Th2 cytokines may result in a reduction in serum IgE levels. Our findings suggest that mechanism of action of Sea Buckthorn in the treatment of AD is associated with a marked anti-inflammatory effect as well as an inhibition of Th2-mediated IgE overproduction via the modulation of biphasic Th1/Th2 cytokines. Such results suggest that topical Sea Buckthorn extract may prove to be a novel therapy for AD symptoms with few side effects.

• 동의생리병리학회지
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• 제28권2호
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• pp.178-185
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• 2014

Leonuri Fructus, a semen of Leonuri Herba, has been used for the treatment of menstrual disorders such as amenorrhea, dysmenorrhea and leukorrhea and for the remedy of hyperemia. The present study was conducted to evaluate the anti-inflammatory effects of the Leonuri Fructus extract (Leonurus japonicus Houtt. EtOH extract; LJE) in vivo and in vitro. In vitro study, the MTT assay for cell viability was conducted to determine the non-cytotoxic concentration of LJE treatment in media. The levels of NO were measured with Griess reagent. Pro-inflammatory cytokines were detected by ELISA method. The inflammation-related proteins of this study were detected by immunoblot anlaysis. The increases of NO production and iNOS expression were detected in LPS-treated cells compared with control, but LJE attenuated the increases of NO and iNOS by LPS. LJE reduced the production of TNF-${\alpha}$ and IL-$1{\beta}$ induced by LPS stimulation. LJE suppresses the signaling pathways of NF-${\kappa}B$ and MAPKs in LPS-induced macrophage cells. In vivo study, carrageenan-induced hind paw acute edematous inflammation rat model was used for evaluation of anti-inflammatory activity of LJE. LJE significantly inhibited the increases of hind paw swelling, skin thicknesses and inflammatory cell infiltrations, and decreased the numbers of mast cell induced by carrageenan injection. These results suggest that LJE has an anti-inflammatory therapeutic potential, which is mediated through modulating NF-${\kappa}B$ activation and MAPK phosphorylation. Inhibition of the rat paw edema induced by carrageenan is considered as direct evidence that LJE may be a useful source to treat inflammation.

• 약학회지
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• 제41권3호
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• pp.389-398
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• 1997

Non-adenergic non-cholinergic (NANC) innervation on the circular muscle of the rabbit gastric body was investigated by observing the magnitudy of relaxations induced by the elec trical field stimulation (EFS). Strips were cut from the greater curvature of the gastric body and stimulated with 5s trains of 0.5 ms pulses at 1-20 Hz, 40 V. The EFS induced transient frequency-dependent contractons, followed by a slowly recovering relaxation ewpecially at higher frequency of the EFS. In the presence of atropine and guanethidine, the contractions were virtually abolished, while the frequency-dependent relaxations by the EFS remained unaffected. The magnitude of relaxations progressively decreased as the location of the strips gets closer to the bottom of the gastric body. The relaxations were ablished by tetrodotoxin, indicating that their orgin is the NANC nerve stimulation. NG-nitro-L-arginine (L-NNA, 10-$100{\mu}M$), the inhibitor of nitric oxide (NO)-synthase, caused a concentration-dependent inhibition of the NANC relaxations. The inhibitory effects of L-NNA were not affected gy the location of the strips and were reversed by L-arginine, the precursor of NO-biosynthesis. Hemoglobin (20-$60{\mu}M$), a NO scavenger, inhibited the NANC relaxation s in a concentration-dependent manner. This inhibition was more prominent in the NANC relaxations observed in the lower portion of the gastric body and the relaxations induced ly lower frequencies of the EFS. Methyelne blue (10-$100{\mu}M$), an inhibitor of cytosolic guanylate cyclase, markedly inhibited the NANC relaxations, almost abolishing the response at a higher dose ($100{\mu}M$). These results suggest that NANX innervation of the rabbit gastric body progeressively decrease as he location of the strips gets closer to the bottom of the gastric body, and that the NANC relaxation is primarily mediated by NO-guanosine 3',5'-cyclic monophophate (cyclic GMP).

• 한국약용작물학회지
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• 제28권1호
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• pp.9-20
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• 2020

Background: Although a number of Panax ginseng cultivars have been developed by Korean researchers in recent years, there has been insufficient analysis of their beneficial properties. In this study, we sought to identify useful ginseng varieties as functional materials. Methods and Results: We evaluated effects of root extracts of 10 ginseng cultivars (Cheongsun; CS, Chunpoong; CP, Gopoong; GP, Gumpoong; GMP, K1, Sunhyang; SH, Sunone; SO, Sunpoong; SP, Sunun; SU and Yunpoong; YP) against the inhibitory effects of nitric oxide (NO) and reactive oxygen species (ROS) production in mouse brain microglial BV2 cells, as well as the binding of N-methyl-D-aspartate receptor (NMDAR), a marker related to memory. Ginsenosides, such as 20 (S)-protopanaxadiols (PPDs), including ginsenoside-Rb1, -Rb2, -Rb3, -Rc, -Rd, and - Rg3 and 20 (S)-protopanaxatriols (PPTs) including -Re, -Rg1, and -Rg2 were analyzed by HPLC. We observed that the cultivar GMP showed the highest inhibitory effect (60.8%) against NO production at 20 ㎍/㎖. Those cultivars showing the significantly highest inhibition effects against ROS at 20 ㎍/㎖ were K1 (57.3%), SP (54.5%), YP (53.1%), CP (51.7%), CS (50.9%) and SH (49.6%). At 50 ㎍/㎖, K1 showed the most potent inhibitory effect (51.2%) on NMDAR binding. The total phenol content of SH (1.89 mg/g) and K1 (1.73 mg/g) were higher than those of the other cultivars, whereas in terms of PD/PT ratios, the values of CP (0.98), K1 (1.05) and SO (1.05) were lower than those of the other cultivars. On the basis of correlation coefficient (0.7064) between NMDAR inhibition and ONOO^- scavenging activity. Conclusions: The findings of this study indicate that the cultivars K1 and SH could be useful ginseng resources as functional materials with favorable cognition-improving and antioxidative properties.

• Journal of Acupuncture Research
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• 제24권6호
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• pp.15-27
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• 2007

Objectives : The purpose of this study is to investigate the effectiveness of Ulmus davidiana Planch herbal acupuncture(UA) to inhibit nuclear $factor(NF)-{\kappa}B$ activation on type II collagen-induced arthritis (CIA) in mice. Methods : Using an in vitro test, the synoviocytes picked out from the experimental CIA mice were subcultured. The synoviocyte cells were treated with phorbol-12-myristate-13-acetate(PMA) for 1 hour prior to the addition of indicated concentrations($0.4\;-\;1.0mg/m{\ell}$) of UA, and the cells were further incubated for 24 hours. The in vivo test, $NF-{\kappa}B$ p65, inducible nitric oxide synthase(iNOS), cyclooxygenase-2(COX-2), vascular cell adhesion molecule(VCAM)-1 production and apoptosis was observed by immunohistochemical staining. Results : The PMA-induced $I{\kappa}B$ kinase(IKK), iNOS and COX-2 mRNA expression were dose-dependently decreased in UA treated synoviocytes. Using the in vivo test, the number of eosinophils in mice treated with UA noticeably decreased in the the CIA group(P<0.05 using student t test). In mice treated with UA, there was less cartilage erosion. less bone destruction, mild synovial hyperplasia, mild fibrosis, and mild angiogenesis with less MIP-2 production. By immunohistochemical staining, suppression of $NF-{\kappa}B$ p65, iNOS production, inhibition of COX-2 production, inhibition of VCAM-1 production and inducing apoptosis were observed. Conclusions : These results suggest that UA might be applicable to the therapy of RA to suppress $NF-{\kappa}B$ activation.