• Title/Summary/Keyword: mRNA 발현

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Effects of Lithospermum erythrorhizon on the cytokine gene expression in human keratinocytes (자초(紫草)가 HaCaT 세포의 사이토카인 유전자 발현에 미치는 영향)

  • Kang, Sang-Hoon;Kim, Gyung-Jun
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.26 no.1
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    • pp.50-62
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    • 2013
  • Objective : Propionibacterium acnes (P. acnes) is a major pathogenic bacteria for acne vulgaris. This study was performed to evaluate the effects of Lithospermum erythrorhizon extracts on the inflammatory cytokines gene expression by P. acnes in human keratinocytes, HaCaT cell line. Methods : Anti-bacterial activity and cytotoxicity of LE extracts was analyzed by agar plate culture and XTT assay. The cytokines gene expressions were assessed by real time RT-PCR for IL-8, MCP-1 and TNF-${\alpha}$. During the cell culture and treatments, amounts of secreted TNF-${\alpha}$ were measured by ELISA. Translocation of transcription factor NF-${\kappa}B$ from cytoplasm into nucleus was observed by immunocytochemistry and confocal microscopy. Results : There were no anti-bacterial effects and cytotoxicity as high as $1,000{\mu}g/ml$ of LE extracts in XTT assay. Transcription levels of inflammatory cytokines, IL-8, MCP-1 and TNF-${\alpha}$ were increased by P. acnes in HaCaT. LE extracts decreased the upregulated gene transcription levels. However, amounts of secreted TNF-${\alpha}$ were similar in HaCaT cells with P. acnes and LE extracts. Translocation of NF-${\kappa}B$ into nucleus by P. acnes was significantly inhibited by LE extracts. Conclusions : From the results of this study, LE extracts have anti-inflammatory effects on HaCaT cells by P. acnes that decreased the mRNA expressions of IL-8, MCP-1 and TNF-${\alpha}$. This anti-inflammatory effects of LE extracts could provide the potential of therapeutic substance for acne vulgaris.

Antioxidant and anti-inflammatory effects of the ethyl acetate fraction of the Agastache rugosa extract (배초향 에틸아세테이트 분획의 산화방지 및 항염증 활성)

  • Kim, Bomin;Han, Yeong Eun;Lee, Hwa Jin
    • Korean Journal of Food Science and Technology
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    • v.49 no.3
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    • pp.331-337
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    • 2017
  • To evaluate the antioxidant activity of hexane, ethyl acetate, and butanol fractions obtained from Agastache rugosa extract, we measured the total polyphenol levels, DPPH radical scavenging activity, and reducing power. The ethyl acetate fraction of A. rugosa (AREA) displayed high phenolic levels, potent DPPH radical scavenging effect, and powerful reducing power. In addition, we examined the ability of AREA to inhibit nitric oxide (NO) production in lipopolysaccharide (LPS)-activated BV-2 microglia. AREA suppressed NO production and inducible nitric oxide synthase (iNOS) expression and downregulated interleukin-6 (IL-6) mRNA level in LPS-stimulated BV-2 microglia. Furthermore, we detected rosmarinic acid in AREA by HPLC, which suggested that rosmarinic acid could be one of the bioactive materials responsible for the antioxidant and anti-inflammatory activities of AREA. These results suggested that AREA may be a good source of functional foods with antioxidant and anti-inflammatory activities.

The Effect of Heparin on Gene Expression of Collagen, MMP, TGF-β Isoforms Using Cultured Fibroblast from Transverse Carpal Ligament of Carpal Tunnel Syndrome (헤파린이 수근관 증후 군 환자의 횡수근 인대에서 추출한 섬유아세포의 콜라겐, MMP 및 TGF-β 동위형 발현에 미치는 영향)

  • Jeong, Seong Ho;Na, Min Hwa;Lee, Jae Sun;Song, Sang Hun;Yoon, Eul Sik;Dhong, Eun Sang;Kim, Han Joong
    • Archives of Plastic Surgery
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    • v.33 no.4
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    • pp.427-432
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    • 2006
  • Purpose: Noninflammatory synovial fibrosis has been noted for main causal factor of carpal tunnel syndrome (CTS). Recently, there are some reports that heparin have not only anti-coagulative effect but also anti-inflammatory and anti-fibrotic potential and have an effect on interstitial pulmonary fiborosis. Authors examined whether heparin affects pathogenesis of CTS. Methods: First, heparin was administered to fibroblast that was cultured from patient's transverse carpal ligament. Secondly, we evaluated the expression from genes of type I, III collagen, TGF ${\beta}$ isoforms and MMP. Fibroblasts were isolated and cultured from transverse carpal ligaments of 5 patients with CTS. Heparin (0, 1, 10,$100{\mu}g/ml$) was administered to cultured fibroblast and reverse transcription PCR for mRNA expression of type I, III collagen, TGF-${\beta}$ isoforms and MMP was done. Results: Heparin suppressed gene expression of type I, III collagen and TGF-${\beta}1$, ${\beta}3$ but promoted gene expression of TGF-${\beta}2$ and MMP-2. Conclusion: Heparin directly suppress gene expression of type I, III collagen. But, It is undetermined that heparin can present it's effect mediated by TGF ${\beta}$ isoforms or MMP.

Expression of protein kinase C in the spinal cords of rats with autoimmune encephalomyelitis (뇌염모델에서 Protein Kinase C의 발현에 관한 연구)

  • Shin, Tae-Kyun;Kim, Hyung-Min;Tanuma, Naoyuki;Matsumoto, Yoh
    • Korean Journal of Veterinary Pathology
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    • v.1 no.1
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    • pp.26-32
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    • 1997
  • Protein kinase C an enzyme of signal transduction has been known to regulate cell proliferation activation as well as apoptosis in some cancer cell lines. To explore the role of PKC in the course of cell mediated autoimmune disease such as experimental autoimmune encephalomyelitis (EAE) EAE was induced in Lewis rats(6-8 weeks old) with immunization of myelin basic protein supplemented with complete Freund's adjuvants and affected spinal cords were sampled at days 13 postimmunization(PI) as peak stage of EAE and at days 21 PI as recovery stage. The spinal cords with EAE were subjected to Northern blot analysis and insitu hybridization of PKC delta which is one of prominant isotypes of PKC in the haematopoietic cells. Northern blot analysis showed that levels of PKS delta mRNA in the spinal cords of rats withEAE was significantly increased at days 13 PI in which inflammatory cells including T cells and macrophages in the EAE lesions appeared. however the stage. By in situ hybridization signals of PKC delta in EAE lesions was intensely expressed on the delta is also expressed on some brain cells in normal rat central nervous system This finding suggests that PKC plays an important role on either activation of inflammatory cells including encephalitogenic T cells and macrophages or apoptotic elimination of some inflammatory cells depending on the stge of EAE.

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GENE EXPRESSION AFTER THE APPLICATION OF THE FLUID-INDUCED SHEAR STRESS ON THE GINGIVAL FIBROBLAST (유체에 의해 유발된 전단력이 치은 섬유아세포 유전자 발현 변화에 미치는 영향에 관한 연구)

  • Jeong, Mi-Hyang;Choi, Je-Yong;Chae, Chang-Hoon;Kim, Seong-Gon;Nahm, Dong-Seok
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.27 no.5
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    • pp.424-430
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    • 2005
  • The oral cavity is humid environment mainly due to the continuous salivary flow. The reaction of oral mucosa to fluid flow is important for homeostasis and pathogenesis. The objective of this study is the screening the change of gene expression after the application of fluid induced shear stress (FISS) on the gingival fibroblast using cDNA microarray assay. The immortalized human gingival fibroblasts were grown and FISS was applied using a cone viscometer at a rotational velocity of 40 rpm, respectively for periods of 2 and 4 hours. The synthesis of cDNA was done from the extracted total RNA and cDNA microarray assay was done subsequently. The genes that showed over 1.6 in the Cy3/Cy5 or the Cy5/Cy3 value were regarded as genes influenced significantly by the FISS application ion (/M/>0.7). The " RUNX-1" was increased its expression in 2 hours group and " RUN and SH3 domain containing 1" was increased its expression in 4 hours group. The "CC020415", "cyclin L1", "interferon regulatory factor1", "early growth response 1", "immediate early response 2", and "immediate early response 3" genes were increased their expression in 2 and 4 hours after FISS application. In conclusion, we could find many genes that were probably related to the FISS application. Interestingly, most of them were placed in similar molecular pathways and these findings improve the reliability of chip data and usefulness in overall screening. From this experiment, we could find many items for further study and it will make improvement in the understanding of intracellular events in response to FISS.

Effect of Kami-Cheongsimyeonjatang on cytokine expression with GATA3 regulation in atopic dermatitis-like skin lesions and IgE hyperproduction induced in NC/Nga mice (IgE 과대생산과 피부염이 유발된 NC/Nga생쥐의 비장세포에서 GATA3 조절에 의한 유전자 발현에 미치는 영향)

  • Park, Seul-Ki;Han, Jae-Kyung;Kim, Yun-Hee
    • Journal of Haehwa Medicine
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    • v.17 no.2
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    • pp.167-183
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    • 2008
  • KCSYJT medicines controlled $CD4^+/IFN-\gamma$, and $CD4^+/CD25^+/foxp3^+$ revelation that an experiment that motive allergy immune reponse because an in vitro experiment stimulates T cells of a NC/Nga mouse same time by anti-CD40/rmIL-4, and interleukin-$1{\beta}$, IL-6, TNF-$\alpha$, and TGF-$\beta$ mRNA outturn that bear in T and B cells decreased remarkably by KCSYJT medicines. Intracellular staining of splenocytes anti-CD40/rmIL-4 plus rmIL-4 stimulated as described in a, assessed after 24 h, KCSYJT exerts a mainly immunosuppressive effect that acts at least partially through suppression of the transcription factor GATA3 expression in $CD4^+$ T cells. We found that skin lesions, which were clinically and histologically very similar to human AD, mite antigen-induced dermatitis on the face, neck, ears and dorsal skin of inbred NC/Nga mice. Result that Th1 cell and Th2 cell observe to be shifted by cytokine expression with GATA3 regulation by KCSYJT medicines could know that KCSYJT medicines can use usefully in allergy autoimmnune diease.

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Isolation and mRNA Expression of Metallothionein Isoforms from Rockbream Oplegnathus fasciatus (돌돔(Oplegnathus fasciatus)으로부터 Metallothionein 유전자 Isoform들의 분리 및 발현 특징 분석)

  • Lee, Sang-Yoon;Nam, Yoon-Kwon
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.44 no.2
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    • pp.126-140
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    • 2011
  • Genetic determinants of two metallothionein isoforms (MT-A and MT-B) were isolated and characterized from the perciform species, rockbream (Oplegnathus fasciatus). Rockbream MT-A and MT-B shared a high degree of homology at amino acid levels with representative orthologs from other perciform species, especially with respect to the conserved cysteine residues. At the genomic level, both MT-A and MT-B genes represent a tripartite structure typical of vertebrate MT genes. However, rockbream MT-B showed unusually large introns (1.2 kb and 0.8 kb for intron I and II, respectively), a phenomenon that has rarely been seen in other vertebrate MT genes. MT-A and MT-B transcripts were ubiquitously detected in a wide array of tissues, wherein brain and eye showed the highest basal expression levels, and the fin exhibited the lowest expression of both isoforms. The basal expression of MT-A in most tissues was significantly higher (ranging from 4- to 10-fold) than that of MT-B. Upon heavy metal exposures to Cd, Cu or Zn at 25 ppb for 48 h, MT-A and MT-B transcripts in the liver were significantly activated by Cd and moderately by Zn. On the other hand, exposure to Cu did not result in alterations of MT-A, nor in the significant suppression of MT-B. Following bacterial challenges with Escherichia coli, Edwardsiella tarda or Streptococcus iniae, MT isoforms in the liver, kidney and spleen were highly modulated and exhibited a pattern that was dependent on the bacterial species, tissues and isoforms. These results suggest that the two MT isoforms could be taken into account as potential indicators of metal toxicity and immune perturbations of this aquaculture-relevant species.

Effect of Ferulic Acid Isolated from Cnidium Officinale on the Synthesis of Hyaluronic Acid (천궁으로부터 분리된 ferulic acid의 히알루론산 생성에 미치는 효과)

  • Song, Hye Jin;Jin, Mu Hyun;Lee, Sang Hwa
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.39 no.4
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    • pp.281-288
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    • 2013
  • Hyaluronic acid (HA) is one of the major extracellular matrix components in skin. The HA content is reported to decline with age, which may contribute to decrease in skin moisture, wrinkle formation and the decrease in elasticity of the skin. Among the family of HA synthase genes (HAS-1, 2, 3) identified so far, HAS-2 plays crucial roles in the regulation of HA synthesis in human skin fibroblasts. In this study, we elucidated the effects of ferulic acid isolated from Cnidium officinale on HA production. Semi-quantitative RT-PCR and quantitative real-time PCR showed that ferulic acid increased mRNA level of HAS-2 gene and ELISA assay also revealed that ferulic acid increased HA production in human skin fibroblasts. Our study suggests that ferulic acid might prevent age-dependent skin deteriorations such as wrinkles, dryness and elasticity decrease, all of which could be ascribed to the reduction of the HA content in human skin.

Antioxidant, Anti-wrinkle and Whitening Activities of Chrysanthemum Zawadskii var. Latilobum and Fermented Rhus verniciflura (구절초와 발효 옻나무의 항산화, 항주름 및 미백활성)

  • Eun, Bong-gi;Park, Min-Jeong;Mun, Jeong-Yun;Choi, Ji-Soo;Im, Jong-Yun;Jang, Tae-Won;Park, Jae-Ho
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2018.04a
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    • pp.75-75
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    • 2018
  • 본 연구는 발효 옻나무와 구절초를 복합소재로 활용하여 화장품 소재로서의 개발 가능성을 확인하고자 항산화, 항주름 및 미백활성을 검증하였다. 구절초와 발효옻나무를 각각 및 각 비율별로 혼합하여 항산화 활성, 항주름 활성, 미백활성을 확인하였다. 구절초와 발효옻나무의 최적 혼합비율을 설정하기 위해 다양한 혼합비에서 선별하였으며, 구절초 1: 옻나무 9의 활성이 가장 효과적이었다. 혼합물의 DPPH, ABTS 라디칼 소거활성은 $1,000{\mu}g/ml$에서 각각 $95.78{\pm}3.24%$, $99.01{\pm}1.80%$로 나타났고, reducing power은 $1,000{\mu}g/ml$에서 $81.48{\pm}1.47%$로 확인되었다. Tyrosinase 저해활성은 $1,000{\mu}g/ml$에서 $16.74{\pm}1.85%$로, Collagenase 저해활성은 $20{\mu}g/ml$에서 $112.40{\pm}7.75%$로 나타났다. 미백 활성을 확인하기 위한 대조군으로 arbutin을 사용하였으며, B16 F10 세포를 통한 미백 활성은 혼합물 처리에 의해 tyrosinase, TRP-1, TRP-2, MITF protein 및 mRNA 발현을 유의성 있게 저해하였으며, arbutin과 유사한 효과를 나타냈다. 또한 wound healing assay를 통한 피부 장벽 손실 억제 효과를 확인하였다. 따라서 본 연구에서는 구절초와 발효옻나무의 혼합물의 높은 항산화, 항주름 및 미백활성을 통해 천연화장품의 소재로서의 높은 가치를 확인하였다.

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Effects of Antioxidant on the Hypoxia-induced Expression of ICAM-1 in Cultured Human Synovial Fibroblasts (저산소증에 의한 활막 섬유모세포의 ICAM-1 발현에 대한 항산화제의 영향)

  • Kim, Jung Ryul;Yoo, Wan Hee
    • IMMUNE NETWORK
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    • v.2 no.1
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    • pp.25-34
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    • 2002
  • Background: Rheumatoid arthritis (RA) is a chronic inflammatory disease characterized by synovial hyperplasia and joint destruction. The synovial fibroblasts express cell adhesion molecules and have a role in adhesive interation with inflammatory cells in synovial tissue. It has been suggested that hypoxic conditioins are thought to exist in arthritic joints, and several studies indicate that reactive oxygen species (ROS) produced in hypoxic condition can initiate events that lead to pro-adhesive changes via increased expression of adhesion molecules. So, this study wsa designed to examine whether antioxidant can inhibit hypoxia-induced expression of ICAM-1 in cultured human synovial fibroblasts. Methods: Synovial fibroblasts were isolated from synovial tissue in patients with RA and cultured at hypoxic condition. Antioxidant, PDTC (pyrrolidine dithiocarbamate) were pre-treated for an hour before the hypoxic culture and synovial fibroblasts were harvested at 0, 6, 12, 24, 48 hours time points. Cell surface ICAM-1 expression in synovial fibroblasts was examined by the flow cytometric analysis. To analyse the expression of ICAM-1 mRNA, reverse-transcriptase polymerase chain reaction (RT-PCR) was performed. The levels of cytokines in culture supernatants were measured by ELISA, and activation of NF-${\kappa}B$ was analysed by electrophoretic mobility shift assay. The adhesive reaction between synovial fibroblasts and lymphocytes was assayed by measurement of fluorescent intensity of BCECF-AM in lymphocytes. Results: Hypoxic stimuli up-regulated the ICAM-1 expression as well as the adhesive interaction of human synvial fibroblasts to lymphocytes in a time-dependent manner, and PDTC inhibited hpyoxia-induced ICAM-1 expression and cell-cell interaction. PDTC also inhibited the hypoxia-induced activation of intracellular transcription factor, NF-${\kappa}B$. PDTC decreased the amount of hypoxia-induced production of IL-$1{\beta}$ and TNF-${\alpha}$. Conclusion: These studies demonstrate that PDTC inhibit the hypoxia-induced expression of the adhesion molecule, ICAM-1 and activation of NF-${\kappa}B$ in cultured human synovial fibroblasts.