• Title/Summary/Keyword: hemin

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Inhibition of NO Production by Ethanol Extract of Prunus mume Fruits in LPS-Stimulated RAW 264.7 Macrophages through Regulation of the Nrf2/HO-1 Signaling Pathway (LPS가 처리된 RAW 264.7 대식세포에서 Nrf2/HO-1 경로 조절을 통한 매실 추출물의 NO 생성 억제 효과)

  • Kang, Hye-Joo;Choi, Eun Ok;Jeong, Jin-Woo;Park, Shin-Hyung;Park, Cheol;Hong, Su Hyun;Shin, Soon Shik;Cheong, Jae-Hun;Choi, Yung Hyun
    • Herbal Formula Science
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    • v.25 no.1
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    • pp.1-10
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    • 2017
  • Objectives : The fruit of Prunus mume Siebold & Zucc. has been used as an alternative medicine and functional food in Korea and Japan for preventive and therapeutic purposes. However, its molecular actions and mechanism on anti-inflammatory activity have not been clearly investigated. The aim of this study was to clarify the anti-inflammatory activity of the ethanol extract of P. mume fruit (EEPM) in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells, and sought to understand the associated molecular mechanisms. Methods : Cytotoxicity was assessed by an MTT assay. The amount of nitric oxide (NO) production was determined by nitrite assay. The mRNA expression of inducible nitric oxide synthase (iNOS) was analyzed by RT-PCR. In addition, expression levels of iNOS, nuclear factor-erythroid 2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) protein were detected by Western blotting. Results : Our data indicated that EEPM inhibited NO production in LPS-stimulated RAW264.7 cells in a concentration-dependent manner. At the mRNA and protein levels, EEPM suppressed LPS-induced iNOS expression. On the other hand, EEPM markedly enhanced HO-1 expression, which was associated with an induction and nuclear translocation of Nrf2. Moreover, the inhibitory effect of EEPM against LPS‑induced NO production was significantly enhanced by hemin, a HO-1 inducer; however, EEPM's effect on the production of NO was abolished by zinc protoporphyrin IX, a HO-1 inhibitor. Conclusion : The results suggest that EEPM can act as a suppressor agent on NO production through an activation of Nrf2/HO-1 signaling pathway, and may be a promising candidate for the treatment of inflammatory diseases.

SEM Study on the Anaerobic Bacterial Adhesion to the Dentin of Root Canal (혐기성 미생물의 근관내 상아질 부착에 대한 주사전자현미경적 연구)

  • Sung-Eun Yang;Kwang-Shik Bae
    • Restorative Dentistry and Endodontics
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    • v.26 no.4
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    • pp.350-359
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    • 2001
  • 목적 - 근관형성시 근관내 상아질벽에는 항상 도말층(smear layer)이 형성되는데, 이는 상아질, 치수조직 잔사, 조상아세포 돌기, 때로는 미생물 등으로 구성되며, 주사전자현미경상으로는 비규칙적이며, 무정형의 구상(granular)구조물로 관찰된다. 본 연구에서는 도말층의 유무에 따른 혐기성 미생물, Prevotella nigrescens의 근관내 상아질 부착정도를 주사전자현미경으로 평가하고자 한다. 지금까지 사용되었던 실험방법에 비하여 보다 임상적 환경에 가까운 실험방법을 고안, 신빙성 있는 연구결과를 기대할 수 있도록 하였다. 방법 - 치주질환이나 외상 등의 원인으로 발거된 상, 하악 전치 18개를 사용하였다. 각 치아의 치관부를 백악법랑경계부위에서 절단하고, 1군(5개치아)은 10ml의 생리식 염수를, 2군(5개치아)과 3군(5개치아)은 10ml의 3.5% NaOCl을 근관관주용액으로 사용하여 근관형성을 하였다. 근관형성 완료후 1군과 2군은 10ml의 생리식염수로, 3군은 10ml의 0.5M EDTA용액으로 final flush를 시행하여, 3군의 도말층을 제거하였다. 치근수직절단과 ethylene oxide(EO) gas 소독후 1, 2, 3군의 시편(각군10개시편)을 Prevotella nigrescens가 부유된 Brain Heart Infusion with Yeast estract, Hemin and Menadione(BHIYHM) broth내에 37$^{\circ}C$에서 3시간 동안 incubation했다. 4, 5, 6군은 실험과정을 검증하기 위한 대조군으로써, 4군(1개치아)과 5군(1개치아)은 1군, 2군과 같이 각각 생리식염수와 NaOCl만을 이용한 근관형성으로 도말층을 잔존시키고, 6군(1개치아)은 3군과 같이 NaOCl과 EDTA를 적용하여 도말층을 제거한 후, 치근 수직절단과 EO gas 소독을 시행했다. 모든 시편(1, 2, 3, 4, 5, 6군)을 통상의 방법에 따라 처리한 후 주사전자현미경을 통하여 관찰, 근관내면에 부착되어 있는 Prevotella nigrescens의 개수, 모양, 상아세관 및 도말층과의 관계 등을 관찰, 비교, 분석하였다. 결과는 t-test와 one-way ANOVA를 통하여 통계처리 하였다. 결론 - 1. 근관형성 후 근관내 상아질 표면 전체는 도말층으로 덮여 있는 양상을 보였다. 2. 3.5% NaOCl과 0.5M EDTA를 적용하여 근관내 도말층을 효과적으로 제거할 수 있었으며, 상아세관 개구부가 확연히 노출되어 있는 소견을 관찰할 수 있었다. 3. 도말층이 덮인군에서 미생물의 부착이 유의성 있게 높았다(P<0.05). 4. 근관 형성중 형성되어 근관 상아질을 덮고 있는 도말층이 미생물의 부착을 증가시켜, 근관 재감염의 기회를 증가시킴을 알 수 있었다.

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Red fluorescence of oral bacteria interacting with Porphyromonas gingivalis (Porphyromonas gingivalis가 일부 구강미생물의 형광 발현에 미치는 영향)

  • Kim, Se-Yeon;Woo, Dong-Hyeob;Lee, Min-Ah;Kim, Ji-Soo;Lee, Jung-Ha;Jeong, Seung-Hwa
    • Journal of Korean Academy of Oral Health
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    • v.41 no.1
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    • pp.22-27
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    • 2017
  • Objectives: Dental plaque is composed of 700 bacterial species. It is known that some oral microorganisms produce porphyrin, and thus, they emit red fluorescence when illuminated with blue light at a specific wavelength of <410 nm. Porphyromonas gingivalis belongs to the genus Porphyromonas, which is characterized by the production of porphyrin. The aim of this study was to evaluate red fluorescence emission of some oral microorganisms interacting with P. gingivalis. Methods: Five bacterial strains (P. gingivalis, Streptococcus mutans, Lactobacillus casei, Actinomyces naeslundii, and Fusobacterium nucleatum) were used for this study. Tryptic soy agar medium supplemented with hemin, vitamin K3, and sheep blood was used as a growth medium. The fluorescence emission of bacterial colonies was evaluated under 405 nm-wavelength blue light using a Quantitative Light-induced Fluorescence Digital (QLF-D) camera system. Each bacterium was cultured alone and co-cultured in close proximity with P. gingivalis. The red/green (R/G) ratio of fluorescence image was calculated and the differences of R/G ratio according to each growth condition were compared using the Mann-Whitney test (P<0.05). Results: Single cultured S. mutans, L. casei and A. naeslundii colonies emitted red fluorescence (R/G ratio=$2.15{\pm}0.06$, $4.31{\pm}0.17$, $5.52{\pm}1.29$, respectively). Fusobacterium nucleatum colonies emitted green fluorescence (R/G ratio=$1.36{\pm}0.06$). The R/G ratios of A. naeslundii and F. nucleatum were increased when P. gingivalis was co-cultured with each bacterium (P<0.05). In contrast, the R/G ratios of S. mutans and L. casei were decreased when P. gingivalis was co-cultured with each bacterium (P=0.002, 0.003). Conclusions: This study confirmed that P. gingivalis could affect the red fluorescence of other oral bacteria under 405 nm-wavelength blue light. Our findings concluded that P. gingivalis has an important role for red fluorescence emission of dental biofilm.

Red fluorescence of oral bacteria is affected by blood in the growth medium (성장배지 혈액 유무가 구강미생물의 적색 형광 발현에 미치는 영향)

  • Jeong, Seung-Hwa;Yang, Yong-Hoon;Lee, Min-Ah;Kim, Se-Yeon;Kim, Ji-Soo
    • Journal of Korean Academy of Oral Health
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    • v.41 no.4
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    • pp.290-295
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    • 2017
  • Objectives: Dental plaque emits red fluorescence under a visible blue light near the ultra-violet end of the light spectrum. The fluorescence characteristics of each microorganism have been reported in several studies. The aim of this study was to evaluate changes in red fluorescence of oral microorganisms that is affected by blood in the culture media. Methods: The gram-positive Actinomyces naeslundii (AN, KCTC 5525) and Lactobacillus casei (LC, KCTC 3109) and gram negative Prevotella intermedia (PI, KCTC 3692) that are known to emit red fluorescence were used in this study. Each bacterium was activated in broth and cultivated in different agar media at $37^{\circ}C$ for 7 days. Tryptic soy agar with hemin and vitamin $K_3$ (TSA), TSA with sheep blood (TSAB), basal medium mucin (BMM) medium, and BMM with sheep blood (BMMB) were used in this study. Fluorescence due to bacterial growth was observed under 405-nm wavelength blue light using the quantitative light-induced fluorescence-digital (QLF-D) device. The red, green, and blue fluorescence values of colonies were obtained using image-analysis software and the red to green ratio (R/G value) and red to total RGB ratio (R/RGB value) were calculated for quantitative comparison. Results: The QLF-D images of the AN, LC, and PI colonies showed red fluorescence in all media, but the fluorescence of all bacteria was reduced in TSA and BMM media, compared with in TSAB and BMMB media. Both the R/G and the R/RGB values of all bacteria were significantly reduced in growth media without blood (P<0.001). Conclusions: Based on this in vitro study, it can be concluded that red fluorescence of oral bacteria can be affected by growth components, especially blood. Blood-containing medium could be a significant factor influencing red fluorescence of oral bacteria. It can be further hypothesized that bleeding in the oral cavity can increase the red fluorescence of dental plaque.

MECHANISM IN ANTIBACTERIAL ACTIVITY OF POLYPHOSPHATES AGAINST PORPHYROMONAS ENDODONTALIS (Porphyromonas endodontalis에 대한 Polyphosphate의 항균기전에 관한 연구)

  • Choi, Sung-Baik;Park, Sang-Jin;Choi, Gi-Woon;Choi, Ho-Young
    • Restorative Dentistry and Endodontics
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    • v.25 no.4
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    • pp.561-574
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    • 2000
  • Poly-P has been used to prevent decomposition of foods and has been shown to have inhibitory effect on the growth of gram positive bacteria. The purpose of this study was to evaluate the effect of poly-P on the growth of Porphyromonas endodontalis, a gram negative obligate anaerobic rod, endodontopathic bacterium. P. endodontalis ATCC 35406 was in BHI broth containing hemin and vitamin K with or without poly-P. Inhibitory effect of each poly-P which was added at the beginning(lag phase) or during(exponential phase) the culture, MIC(minimum inhibitory concentration) was determined by measuring the optical density of the bacterial cell at 540nm. Viable cell counts were measured to determined whether poly-P has a bactericidal effect. Leakage of intracellular nucleotides from P. endodontalis was determined at 260nm and morphological change of P. endodontalis was observed under the TEM(transmission electron microscope). Binding of 32P-labeled poly-P to P. endodontalis was examined. SDS-polyacrylamide gel electrophoresis and zymography were performed to observe the changes in protein and enzyme profiles of P. endodontalis, respectively. The results from this study were as follows : 1. The minimal inhibitory concentration(MIC) of poly-P to P. endodontalis appeared to be 0.04~0.05%. 2. Poly-P added to the P. endodontalis culture during the exponential phase of P. endodontalis was as much effective as poly-P added at the begining of the culture, suggesting that the antibacterial effect of poly-P is not much dependent on the initial inoculum size of P. endodontalis. 3. Poly-P are bactericidal to P. endodontalis, demonstrating the decrease of the viable cell counts. 4. Intracellular nucleotide release from the P. endodontalis, was not increased in the presence of poly-P and was not reversed by the addition of divalent cations like $Ca^{2+}$ and $Mg^{2-}$. 5. Under the TEM, it was observed that fine electro-dense materials were prominent in the poly-P grown P. endodontalis, appearing locally in the cell, and the materials were more abundant and more dispersed in the cell as the incubation time with poly-P increased. In addition, highly electron dense granules accumulated in many poly-P grown cells, most of which were atypical in their shape. 6. Binding of 32P-labeled poly-P to P. endodontalis appeared to be 32.8 and 45.5 and 53.4% at 30 minutes, 1 hours and 2 hours, respectively. 7. In the presence of poly-P. the synthesis of proteins with apparent molecular masses of 25, 27, 35, 45 was lost or drastically decreased whereas expression of a protein with an apparent molecular mass of 75 was elevated. 8. Proteolytic activity of P. endodontalis was decreased by poly-P. The overall results suggest that use of poly-P may affect the growth of P. endodontalis, and the anti-bacterial activity of poly-P seems largely bactericidal. Changes in shape, protein expression, and proteolytic activity of P. endodontalis by poly-P may be directly and indirectly attributed to the antibacterial effect of poly-P. Further studies will be needed to confirm the effect of poly-P.

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Effects of Taeumin, Soeumin and Soyangin Prescriptions on the Adipocyte Induced by Gold Thioglucose in the Rat (태(太)·소음인(少陰人), 소양인(少陽人)의 처방(處方)이 Gold thioglucose로 유발(誘發)된 백서(白鼠)의 비만병(肥滿病)에 미치는 효과(效果))

  • Kim, Kyung-Yo
    • Journal of Sasang Constitutional Medicine
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    • v.8 no.1
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    • pp.295-317
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    • 1996
  • It is researched to elucidate the effects of Taeumjowuitang(TE,太陰調胃湯), Sibimikwanjungtang(SE, 十二味寬中湯) and Yangkeogsanwhatang(SY,凉膈散火湯) on the obesity induced by gold thioglucose and the differentiation and growth of preadipocyte 3T3-L1 in the mouse. The result were as follows: 1. TE,SE and SY extracts improved the blood level of transaminase in the obese mouse induced by gold thioglucose. 2. TE,SE and SY extracts inhibited the increase of liver fat and body fat in the obese mouse induced by gold thioglucose. 3. TE,SE and SY extracts inhibited the increase of body weight in the obese mouse induced by gold thioglucose. 4. TE,SE and SY extracts inhibited the growth of undifferentiate preadipocyte 3T3-L1. 5. TE,SE and SY extracts showed inhibitory effect on the differentiation of preadipocyte 3T3-L1. The above results suggest that the TE,SE and SY extracts may be used on the obesity induced by the overgrowth and differentiation of adipocyte, and the accumulation of fat in liver and body.

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The effects of proliferation and differentiation on adipocyte 3T3-L1 by prescriptions and herbs of Taeyang-In and Taeum-In (태양인(太陽人), 태음인(太陰人)의 처방(處方)과 약재(藥材)가 지방세포(脂肪細胞)(3T3-L1)의 증식(增殖)·분화억제(分化抑制)에 미치는 영향(影響))

  • Kim, Su-beom;Kho, Byung-hee;Song, Il-byung
    • Journal of Sasang Constitutional Medicine
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    • v.10 no.2
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    • pp.533-564
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    • 1998
  • In order to know the effect of proliferation and differentiation on edipocyte 3T3-L1 by prescriptions and herbs, Taeyangin(太陽人)'s Okapijangcheok-tang(五加皮壯脊湯) Mihudeungsikjangtang Acanthopanacis Cortex(五加皮) Phragmitis Rhizoma(蘆根) and Taeumin(太陰人)'s Taeumjowi-tang(太陰調胃湯) Cheongsimyonja-tang(淸心蓮子湯) Cheongpaesagan-tang(淸肺瀉肝湯) Galkeunbupyong-tang(葛根浮萍湯) Coicis Semen(薏苡仁) Rhei Undulati Rhizoma(大黃) Mori Cortex(桑白皮) Ulmi Cortex(楡根白皮) Holotrichia Vermiculus Kalopanaxii Cortex(海桐皮) Ephedrae Herba(麻黃) Imperatae Rhizoma(白茅根), were used and had some effects. 1. The proliferation effect of edipocyte 1) At the Taeyangin(太陽人)'s prescriptions and herbs, Okapijangcheok-tang(五加皮壯脊湯) Mihudeungsikjang-tang Acanthopanacis Cortex(五加皮) have a control effect at the boiling water-extract and ethyl alcohol-extract. Phragmitis Rhizoma(蘆根) have a control effect at the ethyl alcohol-extract. 2) At the Taeyangin(太陽人)'s prescriptions and herbs, Taeumjowi-tang(太陰調胃湯) Cheongsimyonja-tang(淸心蓮子湯) Cheongpaesagan-tang(淸肺瀉肝湯) Galkeunbupyong-tang(葛根浮萍湯) have a control effect at the boiling water-extract and ethyl alcohol-extract. Coicis Semen(薏苡仁) Rhei Undulati Rhizoma(大黃) Morl Cortex(桑白皮) Ulmi Cortex(楡根白皮) Kalopanaxii Cortex(海桐皮) · Ephedrae Herba(麻黃) of the boiling water-extract, Holotrichia Vermiculus Kalopanaxii Cortex(海桐皮) of ethyl alcohol-extract have a control effect on edipocytes. Rhei Undulati Rhizoma(大黃) Ulmi Cortex(楡根白皮) Ephedrae Herba(麻黃) of high-density have a cyto-toxicity. 2. The differentiation effect of edipocyte 1) At the Taeyangin(太陽人)'s prescriptions and herbs during the natural differentiation, Phragmitis Rhizoma(蘆根) of the boiling water-extract, Okapijangchek-tang(五加皮壯脊湯) Acanthopanacis Cortex(五加皮) of the ethyl alcohol-extract have a cyto-toxicity on the first-differentiation. 2) At the Taeumin(太陰人)'s prescriptions and herbs during the natural differentiation, Ulmi Cortex (楡根白皮) Kalopanaxii Cortex(海桐皮) of the boiling water-extract have a cyto-toxicity on the first-differentiation. Cheongsimyonja-tang(淸心蓮子湯) Ephedrae Herba(麻黃) of ethyl alcohol-extract have a control effect on the redifferentiation. 3) At the Taeyangin(太陽人)'s prescriptions and herbs on the first-differentiation during the induced differentiation, Acanthopanacis Cortex(五加皮) of ethyl alcohol-extract has a control effect. Okapijangchek-tang(五加皮壯脊湯) Acanthopanacis Cortex(五加皮) Phragmitis Rhizoma(蘆根) of the boiling water-extract have a cyto-toxicity. 4) At the Taeumin(太陰人)'s prescriptions and herbs on the first-differentiation during the induced differentiation, Coicis Semen(薏苡仁) Ephedrae Herba(麻黃) Imperatae Rhizoma(白茅根) of the boiling water-extract and Ephedrae Herba(麻黃) of the ethyl alcohol-extract have a control effect. Kalopanaxii Cortex(海桐皮) of the boiling water-extract and the ethyl alcohol-extract has a cyto-toxicity. Considering this result, the Taeyangin(太陽人) Taeumin(太陰人)'s prescriptions and herbs have a control effect on edipocytes during the proliferation. Acanthopanacis Cortex(五加皮), Coicis Semen(薏苡仁) Ephedrae Herba(麻黃) Imperatae Rhizoma(白茅根) have a control effect on edipocytes during the induced differentiation. In the future, for treating a obesity need a vivo assay and hope this study to help to know the mechanisms of obesity.

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