• Journal of Ginseng Research
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• 제43권3호
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• pp.394-401
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• 2019

Background: Ginsenoside Rb1, a triterpene saponin, is derived from the Panax ginseng root and has potent antiinflammatory activity. In this study, we determined if Rb1 can increase macrophage phagocytosis and elucidated the underlying mechanisms. Methods: To measure macrophage phagocytosis, mouse peritoneal macrophages or RAW 264.7 cells were cultured with fluorescein isothiocyanate-conjugated Escherichia coli, and the phagocytic index was determined by flow cytometry. Western blot analyses were performed. Results: Ginsenoside Rb1 increased macrophage phagocytosis and phosphorylation of p38 mitogenactivated protein kinase (MAPK), but inhibition of p38 MAPK activity with SB203580 decreased the phagocytic ability of macrophages. Rb1 also increased Akt phosphorylation, which was suppressed by LY294002, a phosphoinositide 3-kinase inhibitor. Rb1-induced Akt phosphorylation was inhibited by SB203580, (5Z)-7-oxozeaenol, and small-interfering RNA (siRNA)-mediated knockdown of $p38{\alpha}$ MAPK in macrophages. However, Rb1-induced p38 MAPK phosphorylation was not blocked by LY294002 or siRNA-mediated knockdown of Akt. The inhibition of Akt activation with siRNA or LY294002 also inhibited the Rb1-induced increase in phagocytosis. Rb1 increased macrophage phagocytosis of IgG-opsonized beads but not unopsonized beads. The phosphorylation of p21 activated kinase 1/2 and actin polymerization induced by IgG-opsonized beads and Rb1 were inhibited by SB203580 and LY294002. Intraperitoneal injection of Rb1 increased phosphorylation of p38 MAPK and Akt and the phagocytosis of bacteria in bronchoalveolar cells. Conclusion: These results suggest that ginsenoside Rb1 enhances the phagocytic capacity of macrophages for bacteria via activation of the p38/Akt pathway. Rb1 may be a useful pharmacological adjuvant for the treatment of bacterial infections in clinically relevant conditions.

• Natural Product Sciences
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• 제25권3호
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• pp.238-243
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• 2019

In this study, the marker compounds of Curcumae Rhizoma (CR) were simultaneously quantified by high-performance liquid chromatography equipped with a photodiode array detector and the anti-inflammatory effects of CR extract and marker compounds in human benign prostatic hyperplasia epithelial-1 (BPH-1) cell lines were investigated. The marker components (4S,5S)-(+)-germacrone-4,5-epoxide, furanodienone, and germacrone, were separated on Gemini $C_{18}$ columns ($250mm{\times}4.6mm$, $5{\mu}m$) at $40^{\circ}C$ by using a gradient of two mobile phases eluting at 1.0 mL/min. Prostaglandin $E_2$ ($PGE_2$) levels in Human BPH-1 cells were determined with an ELISA kit. The coefficients of determination in a calibration curve of each analyte were all 0.9997. The limits of detection and quantification of the three compounds were $0.10-0.32{\mu}g/mL$ and $0.30-0.98{\mu}g/mL$, respectively. The content of three compounds, (4S,5S)-(+)-germacrone-4,5-epoxide, furanodienone, and germacrone, in the CR sample were found to be 5.79 - 5.92 mg/g, 4.72 - 4.86 mg/g, and 1.06 - 1.09 mg/g, respectively. Regarding pharmacological activity against benign prostatic hyperplasia, CR and its components significantly suppressed $PGE_2$ levels of BPH-1 cells. The established analysis method will help to improve quality assessment of CR samples and related products. In addition, CR and its components exhibit antiinflammatory activity in BPH-1 cells, suggesting the inhibitory efficacy of these compounds against the pathogenesis of BPH.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2023년도 임시총회 및 춘계학술대회
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• pp.52-52
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• 2023

Ageing has been known to be deeply related to oxidative activities. Furthermore, inflammation is a response initiated by conditions such as infection and injury. It has been known that complex connections occur between oxidative stress and the inflammatory response, however underlying mechanism needs to be investigated. Chinese resource plants have been widely used as functional food and medicine for a long time, however it is not studied well how many of these resource plants work. We first decided testing anti-oxidant and anti-inflammatory activities with 95% ethanol extracts of the chinese resource plants, Boehmeria pilosinscula (Blume) Hassk., Carlemannia tetragona Hook f. and Clausena emarginata C.C.Huang. To measure anti-oxidant activity, we performed DPPH assay in Raw264.7 cells with 95% ethanol extracts. Clausena extracts showed significantly higher anti-oxidant activity than those of Boehmeria and Carlemannia. We then performed Nitric Oxide assay to measure the inflammation suppression levels with 95% ethanol extracts of these plants. Clausena extracts showed significantly higher suppression of nitric oxide production than those of Boehmeria and Carlemannia. This indicates that inflammation levels are significantly reduced by Clausena. After measuring anti-oxidant and anti-inflammatory activities, we then performed MTT assay to measure the cell survival rate of Raw264.7 cells treated with these extracts. Boehmeria showed much more cell survival rate than Carlemannia and Clausena. Taken together, this result suggests that Clausena extracts have more anti-oxidant and antiinflammatory activities than Boehmeria and Carlemannia, while Boehmeria extracts have more cell survival rate than Carlemannia and Clausena.

• Journal of Ginseng Research
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• 제44권6호
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• pp.790-798
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• 2020

Background: Beneficial effects of Korean Red Ginseng (KRG) on polycystic ovarian syndrome (PCOS) remains unclear. Methods: We examined whether pretreatment (daily from 2 hours before PCOS induction) with KRG extract in water (KRGE; 75 and 150 mg/kg/day, p.o.) could exert a favorable effect in a dehydroepian-drosterone (DHEA)-induced PCOS rat model. Results: Pretreatment with KRGE significantly inhibited the elevation of body and ovary weights, the increase in number and size of ovarian cysts, and the elevation of serum testosterone and estradiol levels induced by DHEA. Pretreatment with KRGE also inhibited macrophage infiltration and enhanced mRNA expression levels of chemokines [interleukin (IL)-8, monocyte chemoattractant protein-1), proinflammatory cytokines (IL-1β, IL-6), and inducible nitric oxide synthase in ovaries induced by DHEA. It also prevented the reduction in mRNA expression of growth factors (epidermal growth factor, transforming growth factor-beta (EGF, TGF-β)) related to inhibition of the nuclear factor kappa-light-chain-enhancer of activated B cell pathway and stimulation of the nuclear factor erythroid-derived 2-related factor 2 pathway. Interestingly, KRGE or representative ginsenosides (Rb1, Rg1, and Rg3(s)) inhibited the activity of inflammatory enzymes cyclooxygenase-2 and iNOS, cytosolic p-IκB, and nuclear p-nuclear factor kappa-light-chain-enhancer of activated B in lipopolysaccharide-induced RAW264.7 cells, whereas they increased nuclear factor erythroid-derived 2-related factor 2 nuclear translocation. Conclusion: These results provide that KRGE could prevent DHEA-induced PCOS via antiinflammatory and antioxidant activities. Thus, KRGE may be used in preventive and therapeutic strategies for PCOS-like symptoms.

• 한국응용과학기술학회지
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• 제34권2호
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• pp.382-393
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• 2017

본 연구에서는 Phytosphingosine(PhS)과 Phytosphingosine-1-phosphate(PhS1P)를 피부 미용 측면에서 생리활성을 연구하여 화장품 소재로서 가능성 여부를 규명하고자 B16F10 멜라닌 세포, RAW264.7 대식 세포, HDF 섬유아세포를 이용하여 3가지 세포에 대한 독성을 파악하고 항염증, 항 멜라닌, MMP-1 발현 억제, Western blot analysis 실험을 통해 효과를 확인 하고자한다. 본 실험 결과 B16F10, RAW264.7, HDF 세포에 대한 독성이 낮은 것으로 확인하였으며, 항염증 NO 저해능 실험에서 PhS1P이 PhS보다 더 강력한 저해활성을 나타냈다. MMP-1 발현 또한 마찬가지로 PhS1P이 더 우수했고, 그 기작은 ERK 활성 감소에 의한 것임을 확인하였다. 한편, 멜라닌 생성 저해능은 알부틴 보다 우수하였으며, 그 작용은 Tyrosinase 발현을 억제하는 것임을 확인하였다. 이상의 결과를 종합하면, 생리활성물질인 PhS과 PhS1P는 주름개선 및 미백화장품의 피부 개선을 위한 기능성화장품으로 활용 가능성을 확인할 수 있었다.

• 한국자원식물학회지
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• 제27권1호
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• pp.29-34
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• 2014

배롱나무 가지가 기능성화장품 소재 및 천연물 의약품으로써 활용 가능성이 있는지 검토해 보고자 그 생리활성을 확인하기 위해서 DPPH radical 소거능 측정, ABTS radical cation decolorization 활성, Nitrite 소거작용, collagenase 저해활성을 측정하였다. DPPH radical 소거능의 경우 배롱나무 가지의 아세톤 추출물은 50 ppm에서 73%의 항산화능을 나타내었으며 합성항산화제인 BHA는 50 ppm에서 90% 이상의 높은 항산화능을 보여준다. ABTS radical cation decolorization 활성은 50 ppm에서 78% 이상 높은 항산화활성을 보였으며 BHA의 경우 50 ppm에서 96% 이상 활성을 보여준다. Nitrite 소거작용 측정은 대조군인 합성항산화제 BHA와 비교하여 모든 농도에서 우수한 아질산염 소거능을 보여주었다. 배롱나무 가지 추출물은 50 ppm의 저농도에서 63% 이상의 활성을 보여주었으며 1000 ppm에서 73%의 활성을 보이는 것과 비교하여 저농도에서도 높은 활성을 보여주는 것을 알 수 있다. Collagenase 저해활성 측정 결과 대조군인 천연물 (-)-epigallo-catechin-3-gallate와 비교하여 모든 농도에서 우수한 collagenase 저해활성을 보여주었으며 50ppm의 저농도에서도 85% 이상의 높은 collagenase 저해활성을 보여주었다. 이와 같은 결과로 미루어 보아 대조군인 인공 합성 항산화제인 BHA와 비교하여 배롱나무 가지 추출물은 항산화 효과, 아질산염 소거작용 그리고 주름개선에 우수한 효과가 나타나는 것을 수 있다. 따라서 화장품 산업 및 천연물 의약품의 원료로 이용하기 위한 적합한 천연 물질인 것으로 판단된다.

• 한국자원식물학회지
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• 제35권5호
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• pp.574-585
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• 2022

순창군에서 생산된 두릅을 40% EtOH을 이용하여 추출한 후 chlorogenic acid 함량, 항산화, 항균, 항염 및 소화효소 활성을 측정하였다. HPLC를 이용하여 chlorogenic acid 함량을 측정한 결과 7.06±0.01 mg/g 함유되어 있었다. DPPH 라디칼 소거활성(SC₅₀)은 4.79±0.05 mg/mL, ABTS 라디칼 소거활성(SC₅₀)은 5.79±0.05 mg/mL, 총 폴리페놀 함량은 170.0±1.8 mgGAE/g, 총 플라보노이드 함량은 105.5±4.1 mgQE/g으로 분석되었다. RAW 264.7 세포, Caco-2 세포에서 세포독성이 나타나지 않았으며, 농도 의존적으로 NO 생성이 억제되었다. RAW 264.7 세포에서 염증성 cytokine인 TNF-α생성은 8.9±0.1 ng/mL, IL-6 생성은 15.2±0.8 ng/mL, IL-1β생성은 30.9±0.9 pg/mL으로 억제되었으며, AEE의 처리 농도가 증가함에 따라 TNF-α, IL-1β, IL-6 생성이 농도 의존적으로 억제되었다. S. typhimurium, L. monocytogenes, H. pylori에 대한 항균활성이 우수하게 나타났으며, 두릅을 EtOH 추출물의 처리농도가 증가함에 따라 효소 활성인 α-amylase와 protease 효소활성도 증가하였다. 순창군에서 생산된 두릅은 chlorogenic acid를 다량 함유하고 있으며, 항산화, 항염, 항균, 소화효소 활성이 우수하게 나타나 향후 건강기능성 소재로 개발할 수 있을 것으로 판단된다.

• 한국식품영양과학회지
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• 제42권2호
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• pp.153-160
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• 2013

감귤 7종을 대상으로 과피를 80% 메탄올로 추출하여 총페놀과 총플라보노이드 함량을 측정하였고, 유해산소 흡수능력(ORAC) 분석을 통한 free radical 소거능, RAW 264.7 murine macrophage를 이용한 NO 생성 저해능, LPS와 t-BHP 자극에 의한 활성산소 생성 저해 활성을 분석하였다. 총페놀 함량은 영귤, 천혜향, 진귤이 각각 30.6, 30.2, 28.2 mg GAE/g으로 높은 값을 나타내었고, 총플라보노이드 함량은 영귤과 진귤이 각각 30.3와 25.5 mg RE/g으로 높은 값을 나타내었다. ORAC 분석을 통한 free radical 소거능은 영귤이 1076 mM TE/g으로 가장 높은 활성을 나타내었으며, 그 다음이 천혜향(1012), 진귤(984), 한라봉(914) 순이었다. 마우스 대식세포인 RAW 264.7에 의한 NO 생성 저해능 ($IC_{50}$)은 천혜향이 215.3 ${\mu}g/mL$로 가장 높았으며, 그 다음으로 영귤(259.2), 진귤(328.9) 순이었다. LPS 자극에 의한 활성산소 생성은 한라봉이 16.4%, 진귤이 12.8% 저해하였고, t-BHP 자극에 의한 활성산소 생성은 진귤, 한라봉, 천혜향이 각각 28.7, 26.1, 26.6% 저해하였다. 총페놀과 ORAC 간의 상관계수는 0.884이고 총플라보노이드와 ORAC 간의 상관계수는 0.855로, ORAC에 의한 항산화 활성은 총페놀 및 총플라보노이드의 함량과 밀접한 관계가 있는 것으로 나타났다. 총페놀 및 총플라보노이드 함량과 NO 생성 저해능 간의 상관계수는 각각 0.921과 0.683으로 NO 생성 저해능도 총페놀 및 총플라보노이드 함량과 밀접한 관계가 있었다.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2018년도 춘계학술발표회
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• pp.14-14
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• 2018

This Camptotheca acuminata Decne. (CA), belonging to Nyssaceae, is a deciduous tree. and has been used as Traditional Chinese medicine since ancient times. The CA produces camptothecin a natural indole alkaloid, and reported to have anti-cancer effects. But the studies on biological activities of CA leaves are insufficient. Therefore, this study confirmed various biological activities such as antioxidant, antidiabetic, anticancer, antiinflammatory and metabolism analysis by HPLC-MS/MS of CA leaves. The $RC_{50}$ values of DPPH radical scavenging activity of ethyl acetate fraction, n-Butanol fraction, methanol extraction, water fraction and n-Hexane fraction were $12.23{\pm}0.01$, $15.93{\pm}0.42$, $55.12{\pm}0.45$, $56.29{\pm}4.15$ and $427.29{\pm}6.13ug/mL$, respectively. The $IC_{50}$ values of ${\alpha}$-glucosidase inhibitory activity of ethyl acetate fraction, n-Butanol fraction, methanol extraction, n-Hexane fraction and water fraction were $24.29{\pm}0.14$, $47.86{\pm}0.45$, $54.23{\pm}1.21$ $466.76{\pm}2.21$ and $623.91{\pm}9.67ug/mL$, respectively. The nitric oxide release activity of n-Hexane fraction, methanol extraction, ethyl acetate fraction, water fraction and n-Butanol fraction were $31.49{\pm}5.74$, $29.79{\pm}0.71$, $26.89{\pm}0.71$, $8.24{\pm}5.83$ and $7.75{\pm}4.08%$ at 25 ug/mL, respectively. The anti-cancer activity of n-Hexane fraction, methanol extraction, ethyl acetate fraction, water fraction and n-Butanol fraction were $31.49{\pm}5.74$, $29.79{\pm}0.71$, $26.89{\pm}0.71$, $8.24{\pm}5.83$ and $7.75{\pm}4.08%$ at 25 ug/mL, respectively. The ethyl acetate fraction activities showed higher biological activities than other fractions. Thus, Additional studies were conducted using ethyl acetate fraction. Metabolite analysis was performed using a LCMS-8040 triple quadrupole mass spectrometer. As a result, Five compounds (1-5) were identified in the ethyl acetate fraction of the CA leave. The identification of these compounds was generated by the analysis of fragmentation methods of the negative and positive ion modes. Five compounds were identified as gallic acid (1), chlorogenic acid (2), isoquercetin (3), astragalin (4) and camptothecin (5). These results suggest that the CA leave can be used for functional materials.

• Journal of Periodontal and Implant Science
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• 제35권1호
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• pp.99-109
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• 2005

It has been focused on the importance of the host inflammatory response in periodontal pathogenesis and progression, treatment has been introduced to control the host response and the method, which diminishes production and activity of MMP by doxycycline, has been used in periodontal field. MMP is a proteolytic enzyme which plays a major role in tissue destruction and MMP-1 is secreted in the periodontally healthy tissue, while MMP-8, 9, 13, etc in the inflammatory state. Among these, MMP-13 has been discovered lately and reported to degrade primarily type II collagen. Periodontal ligament (PDL) cell plays a role in destruction of periodontal tissue. This study was to evaluate the effect of doxycycline and mefenamic acid, non-steroidal antiinflammatory drug on MMP-13 mRNA expression in the rat PDL cell. Doxycycline concentration of $1{\sim}100\;{\mu}g/ml$ was added rat PDL cell and cell activity was measured by MIT assay at day 1 and 3. MMP-13 gene expression was evaluated by RT-PCR after PDL cells were pre-treated for 1hour with doxycycline (50 ${\mu}g/ml$) alone or with mefenamic acid ($10^{-6}M$), then added $IL-1{\beta}$(1.0 ng/ml) and incubated for 16-18 hours. The results are as follows: 1. Cell activity decreased Significantly at 24 and 72 hours in 100 ${\mu}g/ml$ (p<0.05). 2. Level of MMP-13 mRNA was in 20.2% increase by $IL-1{\beta}$ and in pre-treating doxycycline group, expression of $IL-1{\beta}$ induced MMP-13 mRNA was inhibited by 31% than $IL-1{\beta}$ treated only. 3. Mefenamic acid did not inhibit on the expression of $IL-1{\beta}$ induced MMP-13 mRNA, while mefenamic acid in combination with doxycycline inhibited the expression by 41% compared to only $IL-1{\beta}$ stimulation. These results suggest that doxycycline synergistically inhibit the expression of $IL-1{\beta}$ induced MMP-13 mRNA in combination with mefenamic acid.