• 제목/요약/키워드: pPMA

검색결과 172건 처리시간 0.027초

A Cell-Based Assay System for Monitoring NF-$\kappa$B Activity in Human Epidermal Keratinocytes: A Screening Tool of the Antioxidants and Anti-inflammatories for Dermatological Purpose

  • Moon, Ki-Young;Hahn, Bum-Soo;Lee, Jinseon;Kim, Yeong-Shik
    • Journal of the Society of Cosmetic Scientists of Korea
    • /
    • 제27권1호
    • /
    • pp.17-27
    • /
    • 2001
  • A cell-based assay system for monitoring NF-$textsc{k}$B activity was developed to determine the influence of activated NF-$textsc{k}$B in human HaCaT cells. The pNF-$textsc{k}$B-SEAP-NPT plasmid that permits expression of the secreted alkaline phosphatase (SEAP) reported gene in response to the NF-$textsc{k}$B activity and contains neomycin phosphotransferase (NPT) gene for the geneticin resistance in host cells was constructed and transfected into human keratinocyte cell line HaCaT. Human HaCaT transfectant cells secreted the SEAP enzyme into the culture medium in a time-dependent manner until 72h. NF-$textsc{k}$B activities were measured in the SEAP reporter gene assay using a fluorescent detection method. The treatment of HaCaT cell transfectants with known antioxidants [e.g., N-acetyl-L-cysteine and vitamin C] showed inhibition of NF-$textsc{k}$B activity in a time-and concentration-dependent manner. The phorbol 12-myristate 13-acetate (PMA) known as a stimulator of NF-$textsc{k}$B expression demonstrated that it increased NF-$textsc{k}$B activity in a time- and concentration-dependent manner. This assay system could be used to determine the quantitative measurement of NF-$textsc{k}$B activity in the human skin and allow the screening of anti-inflammatory agents from various synthetic chemicals and natural products for dermatological purpose. Abbrevitions used: NF-$textsc{k}$B, nuclear factor kappa B; I-$textsc{k}$B, Inhibitory kappa B; SEAP, secreted alkaline phosphatase; NPT, neomycin phosphotransferease; PCR, polymerase chain reaction: dNTP, deoxynucleoside triphosphates; DMEM, dulbecco’s modified eagle medium; FBS, fetal bovine serum; PBs, phosphate-buffered saline; MUP, 4-methylumbellifery phosphate; NAC, N-acetyl-L-cysteine; DMSO, dimethyl sulfoxide; PMA, phorbol 12-myristate 13-acetate.

  • PDF

Experimental studies on the Whitening and Anti-allergic effect of various Herbal medicines that clear heat (수종의 청열약물의 미백 및 항알러지 효과에 대한 실험적 연구)

  • Lee, Myoung-Gyu;Kim, Kyung-Jun
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
    • /
    • 제20권3호
    • /
    • pp.71-81
    • /
    • 2007
  • Objective : In this study, herbal medicine(GJE, Gardenia jasminoides Ellis; HCT, Houttuynia cordata Thunb.; CIL, Chrysanthemum indicum Linne; PMS,Paeonia moutan Sims, P. subfruticosa Makino; APL, Agrimonia pilosa Ledebour) were screened for their inhibitory activities against Tyrosinase and PMA plus A23187-induced $TNF-{\alpha}$, IL-6, IL-8 productions in HMC-1 cells to reveal their skin -whitening and anti-allergic effect. Method : To investigate Tyrosinase inhibition we treated Mushroom Tyrosinase(Fluka, 93898) $10{\mu}{\ell}$ and 7.5mM Tyrosine (Sigma, T3754) $20{\mu}{\ell}$ with 80% ethanol medicine extracts. Then we observed 96well micro plate extinction at 490nm. In the next experiment, to investigate Anti-allergic effect we blended cultured Human Mast Cells(HMC-1) with medicine extracts. We treated the blended solution with Phorbol 12-myristate 13-acetate(PMA) and A23187, then observed $TNF-{\alpha}$, IL-6, IL-8 by ELISA (enzyme-linked immunosorbent assay) at 450nm. Results : In inhibiting Tyrosinase the results are as follows. 1. We observed 22% inhibition of Mushroom Tyrosinase at $500{\mu}g/m{\ell}$ concentration of GJE extracts. 2. We also could observe that the decreased Mushroom Tyrosinase activities in HCT, CIL extracts. In inhibiing $TNF-{\alpha}$, IL-6, IL-8 productions in HMC-1 cells the results are as follows. 1. Of the extracts examined, HCT, PMS, APL extracts showed over 50% inhibitions of Cytokines at $200{\mu}g/m{\ell}$ concentration. 2. In particular, APL extracts showed the best inhibitory effect on Cytokine productions in a dose-dependent manner. Conclusion : These results suggest that GJE extracts contributes to the anti melanin activities and represent a potential source of whitening agent. Thus these herbal medicines suggest novel drugs on anti-allergic effects.

  • PDF

The Significance of Maturation Score of Brain Magnetic Resonance Imaging in Extremely Low Birth Weight Infant (초극소 저체중 출생아의 뇌 MRI 상 Maturation Score의 의의)

  • Song, In-Gu;Kim, Su-Yeong;Kim, Cur-Rie;Kim, Yoon-Joo;Shin, Seung-Han;Lee, Seung-Hyun;Lee, Jae-Myoung;Lee, Ju-Young;Kim, Ji-Young;Sohn, Jin-A;Lee, Jin-A;Choi, Chang-Won;Kim, Ee-Kyung;Cheon, Jung-Eun;Kim, Woo-Sun;Kim, Han-Suk;Kim, Byeong-II;Kim, In-One;Choi, Jung-Hwan
    • Neonatal Medicine
    • /
    • 제18권2호
    • /
    • pp.310-319
    • /
    • 2011
  • Purpose: The aim of this study was to investigate the effect of perinatal risk factors on brain maturation and the relationship of brain maturation and neurodevelopmental outcomes with brain maturation scoring system in brain MRI. Methods: ELBWI infants born at the Seoul National University Children's Hospital from January 2006 to December 2010 were included. A retrospective analysis was performed with their medical record and brain MR images acquired at near full term. We read brain MRI and measured maturity with total maturation score (TMS). TMS is a previously developed anatomic scoring system to assess brain maturity. The total maturation score was used to evaluate the four parameters of maturity: (1) myelination, (2) cortical infolding, (3) involution of glial cell migration bands, and (4) presence of germinal matrix tissue. Results: Images from 124 infants were evaluated. Their mean gestational age at birth was 27.1${\pm}$2.1 weeks, and mean birth weight was 781.5${\pm}$143.9 g. The mean TMS was 10.8${\pm}$2.0. TMS was significantly related to the postmenstrual age (PMA) of the infant, increasing with advancing postmenstrual age (P<0.001). TMS showed no significance with neurodevelopmental delay, and with brain injury, respectively. Conclusion: TMS was developed for evaluating brain maturation in conventional brain MRI. The results of this study suggest that TMS was not useful for predicting neurodevelopmental delay, but further studies are needed to make standard score for each PMA and to re-evaluate the relationship between brain maturation and neurodevelopmental delay.

Plaque Removal Effect & Gingival Effect by use of the Dentifrice with Di-methyl-s-curetin and NaCl (3% 쑥염 및 0.2% 인진호엑스를 함유한 치약의 치면세균막 제거효과와 치은염 완화효과에 관한 임상실험연구)

  • Shin, Seung-Chul;Suh, Hyun-Seok;Hong, Jee-Suk;Min, Hee-Hong
    • Journal of Korean society of Dental Hygiene
    • /
    • 제1권1호
    • /
    • pp.133-142
    • /
    • 2001
  • The authors have studied the 8 weeks clinical examination On 32 patients of age 20s for experimental group and 34 patients of the same ages for control group, in order to find out the effect of plaque removal effect and gingival effect by use of the dentifrice contained with the Artemisiae Capillaris extract(dimethylesculetin) and NaCl. The obtained results are as followings: 1. Plaque removal effect is better at the experimental group than at the control group on 2 and 8 weeks(p<0.05). 2. There are not significantly differences on the calculus index and stain index between the experimental group and control group, during the 8 weeks experiment(p>0.05). 3. At the 8 weeks experiment, it revealed the less average scores on the PMA index for experimental group, than that of the control group(p<0.05).

  • PDF

Effect of Soybean Fallen Leaves Ethanolic Extract on Expression of Proteins Related to Antioxidant Activity and Cell Invasion (항산화 및 암전이 관련 단백질의 발현에 미치는 콩잎낙엽 에탄올 추출물의 영향)

  • Song, Chaeeun;Lee, Su-Gyeong;Hong, Sugyeong;Ryu, Zoon Ha;Kim, Moon-Moo;Oh, Yunghee
    • Journal of Life Science
    • /
    • 제26권4호
    • /
    • pp.439-445
    • /
    • 2016
  • Soybean leaves, a Korean edible plant material, have been reported to prevent the development of osteoporosis and breast cancer. Based on this rational, soybean fallen leaves ethanolic extract (SBFL) was used for the experiment of cell invasion related to metastasis and antioxidant activity. The effect of SBFL on matrix metalloproteinases (MMPs) in human fibrosarcoma cells, HT1080 as well as its anti-oxidant activity was investigated in this study. The effect of SBFL on scavenging activity of reactive oxygen species was evaluated in vitro using lipid peroxidation assay,DPPH radical and reducing power assay. SBFL showed the positive effects on antioxidant activity, compared with vitamin C and vitamin E used as positive controls. Furthermore, SBFL showed cytotoxicity above 16 µg/ml in MTT assay. In particular, it was found that SBFL decreased the activation of MMP-9 stimulated by phorbol 12-myristate 13-acetae (PMA) and phenazine methosulfate (PMS). SBFL treatment increased the expression levels of p-FoxO-1 and SOD-1. Moreover, SBFL inhibited cell invasion stimulated by vascular endothelial growth Factor (VEGF). These results indicate that SBFL could inhibit cell invasion related to the activation of MMP-9 and oxidative stress, suggesting that it could be available as a main ingredient for prevention of metastasis.

The Effectiveness of Ulmus Davidiana Planch Herbal Acupuncture to Inhibit $NF-{\kappa}B$ Activation on Type II Collagen-induced Arthritis in Mice (유근피(楡根皮) 약침의 $NF-{\kappa}B$ 활성 억제능이 생쥐의 Type II Collagen 유발 관절염에 미치는 영향)

  • Lee, A-Ram;Byun, Hyuk;Park, In-Shik;Jung, Chan-Yung;Kang, Min-Joo;Kim, Eun-Jung;Lee, Seung-Deok;Kim, Kap-Sung
    • Journal of Acupuncture Research
    • /
    • 제24권6호
    • /
    • pp.15-27
    • /
    • 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.

  • PDF

Inhibitory Effect of Dendrobium moniliforme on Degranulation and Histidine Decarboxylase Expression in RBL-2H3 Cells (RBL-2H3 세포에서 탈과립과 histidine decarboxylase 발현에 미치는 석곡(Dendrobium monilifrme)의 효과)

  • Young Ji Lee;Iskander Madhi;YoungHee Kim
    • Journal of Life Science
    • /
    • 제33권2호
    • /
    • pp.176-182
    • /
    • 2023
  • The stems of Dendrobium moniliforme are used in traditional Oriental medicine as a Yin tonic to nourish the stomach, promote the production of body fluid, and reduce fever. This study investigated the effects of the aqueous extract of D. moniliforme stems (DME) on mast cell degranulation and the expression of tumor necrosis factor-α (TNF-α), interleukin-4 (IL-4), and histamine-synthesizing enzyme histidine decarboxylase (HDC). We used rat mast cell line RBL-2H3 cells and stimulated them with PMA plus calcium ionophore (PMACI). Pretreatment with DME significantly inhibited PMACI-induced β-hexosaminidase release and the expression of TNF-α, IL-4, and HDC. Furthermore, DME suppressed PMACI-induced nuclear translocation of the nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) and activator protein 1 (AP-1). In addition, HDC expression was inhibited by SP600125 (JNK inhibitor), PD98059 (ERK inhibitor), and SB203580 (p38 kinase inhibitor). Finally, the phosphorylation of p38 kinase, extracellular signal-regulated kinase 1/2 (ERK1/2), and c-Jun N-terminal kinase (JNK) was inhibited by pretreatment with DME. These results suggest that DME has inhibitory effects against degranulation, cytokine (TNF-α and IL-4) and HDC expression, and that HDC expression is mediated by MAPK signaling. These findings suggest that DME may have therapeutic potential in the treatment of hypersensitive and inflammatory diseases.

Influence on the Indoor Air Quality by Ambient Air during the Summer Season (여름철 실외 공기가 실내 공기질에 미치는 영향)

  • 이학성;강병욱
    • Journal of Environmental Science International
    • /
    • 제6권6호
    • /
    • pp.637-644
    • /
    • 1997
  • The purpose of thins study was to quantitatively determine the Indoor Infiltration of pollutants of outdoor origin. The relationship between Indoor and outdoor air is dependent, to a large extent. on the rate of k exchange between these two environments. Mean Indoor/outdoor ratios measured from thins study were: 0.70 for HNO3; 1.60 for HNO2: 0.56 far SOg: 1.30 for mf3: 0.96 for PM2.5(dP<2.5mm: 0.89 for SO4a': 0.87 for NO3· and 0.79 for NH4 'Mean Indoor concentrations for PMa s, SO4a., HNO9, NO3 and NIL' were similar to outdoor levels. Indoor HNO2 and mB3 values were h19her than outdoors. However, the Indoor level of SO2 was lower than ambient level.

  • PDF

Studies on Anti-Mcirobial and Anti-Cancer Functions of Polysaccharide Extracted from Ganoderma lucidum (영지버섯 다당체의 항미생물작용 및 항암작용에 관한 연구)

  • 김성환
    • Journal of the Korean Society of Food Science and Nutrition
    • /
    • 제27권6호
    • /
    • pp.1183-1188
    • /
    • 1998
  • This study was carried out to elucidate the immunomodulating activity of protein bound polysac charide extracted from Ganoderma lucidum(PSG). Macrophage drived reactive radicals were known as an effector for antimicrobial and anticancer functions. The promising immune response molecule, reactive oxygen intermediates(ROIs), was determined in TIB 71 cells with PSG at various experimental conditions. Treatment with 0.5mg PSG significantly increased the production of ROIs, superoxide anion as well as hydrogen peroxide, from TIB 71 cells(p<0.001). Under the same concentration, con siderable results were obtained from 24 hour cultivation with 106 cells at 5% CO2 incubator. The cells were trigged with PMA(5.3 M) after primed BCG(100 M) or IFN (100U) alone could not induce the production of ROIs, but it had a significant potentiating effect on ROIs secretion when the cells were treated with PSG.

  • PDF