• Title/Summary/Keyword: HUVEC cell

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Photodynamically induced endothelial cell injury and neutrophil-like HL-60 adhesion

  • Takahashi, Miho;Nagao, Tomokazu;Matsuzaki, Kazuki;Nishimura, Toshihiko;Minamitani, Haruyuki
    • Journal of Photoscience
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    • v.9 no.2
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    • pp.518-520
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    • 2002
  • Photodynamic therapy (PDT) is a treatment modality based on photochemical reaction and the resultant cytotoxic reactive oxygen species. The platelet thrombus formation leading to stasis observed in vivo during PDT is called vascular shut down (VSD) effect. To investigate the mechanism of the VSD effect, we observed Human Umblical Vein Endothelial Cell (HUVEC) injury induced by photochemical reaction. We observed cell retraction and blebbing after PDT. It seems that the injury was not fetal and only morphological change. Then, the cytoplasm was stained by Calcein-AM and subendothelial area was evaluated from fluorescence microscopy. The rate of subendothelial area after PDT increased significantly. Second, we investigated interaction between neutrophils and HUVEC. Human promyelocytic leukemia cells (HL-60) were differentiated into neutrophil by incubation with all-trans retinoic acid. Calcein-AM labeled neutrophil adhesion to HUVEC was evaluated from fluorescence microscopy. PDT-induced neutrophil adhesion to HUVEC depended more on the exposure of subendothlial area than on neutrophil activation. This result suggests that there is a certain interaction between neutrophil and HUVEC during PDT.

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The Effect of Gamidohongsamul-tang (GDT) on the Gene Expression Levels of eNOS, KLF2, ICAM-1 and VCAM-1 in HUVEC Cells (가미도홍사물탕(加味桃紅四物湯)이 HUVEC cell 내에 eNOS, KLF2, ICAM-1, VCAM-1의 유전자 발현양에 대해 미치는 영향)

  • Lim, Hyun-chan;Jeon, Sang-yoon
    • The Journal of Internal Korean Medicine
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    • v.39 no.1
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    • pp.1-8
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    • 2018
  • Objectives: This study seeks to investigate the effects of Gamidohongsamul-tang (GDT) on the Gene expression levels of eNOS, KLF2, ICAM-1 and VCAM-1 in HUVEC cells. Methods: HUVEC cells were treated at a concentration of 1, 10, 100 (${\mu}g/ml$) of Gamidohongsamul-tang (GDT). To measure the NOS, KLF2, ICAM-1 and VCAM-1 gene expression in HUVEC cells, the synthesized cDNA was subjected to polymerase chain reaction (PCR) and electrophoresis was performed to verify gene expression level. Results: 1. GDT significantly increased eNOS and KLF2 gene expression. 2. GDT significantly reduced ICAM-1 and VCAM-1 gene expression. Conclusions: These experiments suggest that Gamidohongsamul-tang (GDT) regulates gene expression related with anti-dyslipidemic effects in HUVEC cells. In order to clinically apply this to diseases related to dyslipidemia, such as cardiovascular disease, additional in vivo experiments are needed to verify the anti-dyslipidemic effects of GDT.

The impact of Caesalpinia Sappan L. on Oxidative Damage and Inflammatory Relevant Factor in RAW 264.7 Cells and HUVEC (소목(蘇木)이 산화적 손상 및 Raw 264.7 cell과 HUVEC에서의 염증 유관 인자에 미치는 영향)

  • Kang, Seong-Sun;Kim, Myung-Sin;Jo, Jae-Jun;Choi, Seong-An;Yang, Eui-Ho;Jeon, Sang-Yun;Choi, Chang-Won;Hong, Soek
    • The Journal of Internal Korean Medicine
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    • v.34 no.1
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    • pp.100-111
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    • 2013
  • Objectives : This study investigated the impact of Caesalpinia sappan L. on oxidative damage and inflammatory relevant factor in RAW 264.7 cells and human umbilical vein endothelial cells (HUVEC). Methods : We determined whether fractionated EtOH extracts of Caesalpinia sappan L. (CSL) inhibit free radical generation such as 2,2-diphenyl-1-picrylhydrazyl (DPPH), reactive oxygen species (ROS) and nitric oxide (NO) and pro-inflammatory cytokines in lipopolysaccharide (LPS)-treated RAW 264.7 cells and HUVEC. Result : 1. DPPH removal capacity was increased by CSL. 2. LPS-induced ROS, and NO inhibitory capacity were increased by CSL. 3. LPS-induced cell death of Raw 264.7 cells was decreased by CSL. 4. The amount of cytokine generation in Raw 264.7 cell was decreased significantly by CSL. 5. The amount of cytokine generation in HUVEC was decreased significantly by CSL. Conclusions : These results suggest that CSL supplement may attenuate oxidative stress by elevated antioxidative processes, and suppress inflammatory mediator activation.

Culture of Endothelial Cells by Transfection with Plasmid Harboring Vascular Endothelial Growth Factor

  • Chang, Sungjaae;Sohn, Insook;Park, Inchul;Sohn, Youngsook;Hong, Seokil;Choe, Teaboo
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.5 no.2
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    • pp.106-109
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    • 2000
  • Vascular endothelial cells (EGs) are usually difficult to culture to culture in a large scale because of their complicated requirements for cell growth. As the vascular endothelial growth factor (VEGF) is a key growth factor in the EC culture, we transfected human umbilical vein endothelial cells (HUVEC) using a plasmid containing VEGF gene and let them grow in a culture medium eliminated an important supplement, endothelail cell growth supplement(ECGS). The expression of VEGF by HUVEC tansfected with Vegf GENE was not enough to stimulate the growth of HUVEC, only 40% of maximum cell density obtainable in the presence of ECGS. However, when the culture medium was supplied with 2.5 ng/ml of basic fibroblast growth factor (bFGF), a synergistic effect effect of VEGE and bFGF was observed. In this case, the final cell density was recovered was recovered up to about 78% of maxium value.

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DNA Microarray Analysis of the Gene Expression Profile of Activated Human Umbilical Vein En-dothelial Cells. (올리고 마이크로어래이를 이용한 활성화된 인간 제대 정맥 내피세포의 유전자 발현 조사)

  • 김선용;오호균;이수영;남석우;이정용;안현영;신종철;홍용길;조영애
    • Journal of Life Science
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    • v.14 no.5
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    • pp.874-881
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    • 2004
  • Angiogenesis has been implicated in progression of inflammation, arthritis, psoriasis, atherosclerosis as well as tumor growth and metastasis. Intensive studies have been carried out to develop a strategy for cancer treatment by blocking angiogenesis. During angiogenesis, endothelial proliferation and migration essentially occurs upon activation. In this study, we compared the expression profiles of human umbilical endothelial cells activated by incubating in vitro in the rich medium containing several growth factors, and non-activated ones. cDNA targets derived from total RNAs of HUVEC activated for 13 h in M199 medium containing endothelial cell growth supplement, 20% fetal bovine serum, and heparin, after reaching 70~80% confluency, or non-activated, were hybridized onto oligonucleotide microarrays containing 1,8864 genetic elements. Unsupervised hierarchical clustering analysis resulted in two subgroups on dendrogram exhibiting activated and non-activated HUVECs. We then extracted 122 outlier genes which were shown to be up-regulated or under-expressed by at least 2-folds in activated HUVECs. Among these, 32 annotated genes were up-regulated and 38 were down-regulated in activated HUVECs. Interestingly, genes involved in cell proliferation, motility, and inflammation/ immune response were up-regulated in activated HUVEC, whereas genes for cell adhesion or vessel morphogenesis/function were down-regulated. Unexpectedly, the expression of genes well-characterized as angiogenesis markers was not changed except Eph-B4, which was down-regulated about 4 folds. 52 unknown genes were also up- or down-regulated. Therefore, these results could provide an opportunity to targeting new vascular molecules for the development of anti-angiogenic molecules.

Suppressive Effects of Ethyl Acetate Fraction from Green Tea Seed Coats on the Production of Cell Adhesion Molecules and Inflammatory Mediators in Human Umbilical Vein Endothelial Cells (Human Umbilical Vein Endothelial Cells에서 녹차씨껍질 에틸아세테이트 추출물의 세포부착물질 및 염증매개인자 생성 억제효과)

  • Noh, Kyung-Hee;Kim, Jong-Kyung;Song, Young-Sun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.5
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    • pp.635-641
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    • 2011
  • Anti-atherogenic effects in tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$)-stimulated human umbilical vein endothelial cells (HUVEC) are involved with suppressed oxidative stress, cell adhesion molecules, and pro-inflammatory factors. The aim of this study was to determine whether green tea seed coat ethyl acetate fraction (GTSCE) could modulate cell adhesion molecules and inflammatory mediators in HUVEC stimulated with TNF-${\alpha}$. Nitric oxide (NO) production was significantly increased in TNF-${\alpha}$-stimulated HUVEC compared to TNF-${\alpha}$ only treated cells. The NO that is produced by endothelial nitric oxide synthase dilates blood vessels and has protective effects against platelet and leucocyte adhesion. GTSCE at 25, 50, 75, and $100\;{\mu}g$/mL significantly (p<0.05) reduced TNF-${\alpha}$ production. GTSCE significantly (p<0.05) inhibited soluble vascular cell adhesion molecule-1 level, in a dose-dependent manner. Monocyte chemoattractant protein-1 level was also significantly (p<0.05) inhibited by GTSCE treatment at $75\;{\mu}g$/mL compared to the TNF-${\alpha}$-only treated group. Total antioxidant capacity by GTSCE was significantly (p<0.05) enhanced compared to the TNF-${\alpha}$-only treated group. These results suggest that GTSCE can inhibit the production of cell adhesion molecules and inflammatory mediators and could be used as a candidate bioactive material to prevent the development of atherosclerosis.

Protective Effects of Jihwangeumja on Oxidative Stress-induced Injury of Human Umbilical Vein Endothelial Cells (혈관내피세포의 산화적 손상에 대한 지황음자의 방어기전 연구)

  • 정용준;장재호;이대용;이민구;전인철;정대영;이인;신선호;문병순
    • The Journal of Korean Medicine
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    • v.25 no.2
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    • pp.173-183
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    • 2004
  • Objectives : Oxidative stress can induce negative responses such as growth inhibition or cell death by necrosis or apoptosis due to the intensity of the oxidative stress, as well as positive responses such as cellular proliferation or activation. We examined the effect of Jihwangeumja on this process. Methods and Results : We analyzed the influence of oxidative stress and agents that modify its effect in human umbilical vein endothelial cell (HUVEC). Oxidative stress was induced by $B_2O_2$. With induced oxidative stress the results obtained indicate that it has a harmful effect over cell function and viability, and that this effect is dose and time dependent. When oxidative stress increased, Jihwangeumja reduced cell damage and had protective functions. $B_2O_2$, induced the apoptosis of HUVEC through the activation of intrinsic caspases pathway as well as mitochondrial dysfunction. A significant increase in cell survival was observed in culture cells with oxidative stress when they were treated with Jihwangeumja. Conclusions : These results suggest that Jihwangeumja may be potentially useful to treat HUVEC against oxidative damages mediated by modulation of caspase protease and mitochondrial dysfunction.

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Manassantin A and B Isolated from Saururus chinensis Inhibit $TNF-{\alpha}-Induced$ Cell Adhesion Molecule Expression of Human Umbilical Vein Endothelial Cells

  • Kwon Oh Eok;Lee Hyun Sun;Lee Seung Woong;Chung Mi Yeon;Bae Ki Hwan;Rho Mun-Chual;Kim Young-kook
    • Archives of Pharmacal Research
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    • v.28 no.1
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    • pp.55-60
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    • 2005
  • Leukocyte adhesion to the vascular endothelium is a critical initiating step in inflammation and atherosclerosis. We have herein studied the effect of manassantin A (1) and S (2), dineolignans, on interaction of THP-1 monocytic cells and human umbilical vein endothelial cells (HUVEC) and expression of intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), and E-selectin in HUVEC. When HUVEC were pretreated with 1 and 2 followed by stimulation with $TNF-{\alpha}$, adhesion of THP-1 cells to HUVEC decreased in dose-dependent manner with $IC_{50}$ values of 5 ng/mL and 7 ng/mL, respectively, without cytotoxicity. Also, 1 and 2 inhibited $TNF-{\alpha}-induceda$ up-regulation of ICAM-1, VCAM-1 and E-selectin. The present findings suggest that 1 and 2 prevent monocyte adhesion to HUVEC through the inhibition of ICAM-1, VCAM-1 and E-selectin expression stimulated by $TNF-\alpha$, and may imply their usefulness for the prevention of atherosclerosis relevant to endothelial activation.

Co-Culture Model Using THP-1 Cell and HUVEC on AGEs-Induced Expression of Cytokines and RAGE (THP-1 Cell과 HUVEC을 이용한 Co-Culture Model System에서 최종당화산물에 의한 Cytokines와 RAGE 발현)

  • Lee, Kwang-Won;Lee, Hyun-Sun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.3
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    • pp.385-392
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    • 2011
  • Although monoculture methods have been remarkably useful due to their simplicity, they have serious limitation because of the different types of cells communication with each other in many physiological situations. We demonstrated levels of markers of endothelial dysfunction such as tumor necrosis factor-$\alpha$ (TNF-$\alpha$) and interleukin-1$\beta$ (IL-1$\beta$) as well as stimulation of receptor of advanced glycation endproducts (AGEs) on monoand co-culture system such as only monocyte (THP-1) cultivation system, only endothelial cell (HUVEC) cultivation system, and co-cultivation system of THP-1 and HUVEC. The mRNA levels of TNF-$\alpha$ and IL-1$\beta$ on HUVEC increased by the co-culture with monocyte after 4 hr at 100 ${\mu}g/mL$ glyceraldehyde-AGE. The secreted protein contents into medium of TNF-$\alpha$ and IL-1$\beta$ increased after 8 hr approximately 2~2.5 times compared to mono-cultivation. In contrast, the mRNA level of receptor of AGE (RAGE) was relatively insensitive on the co-culture system. The mediators by which monocytes activate endothelial cell have not been fully elucidated. In this study we confirmed production of soluble cytokines such as TNF-$\alpha$ and IL-1$\beta$ by monocytes. Use of monocyte conditioned medium, which contains both cytokines, can activate endothelial cell.

Inhibitory Effects of Licochalcone A and Isoliquiritigenin on Monocyte Adhesion to TNF-$\alpha$-activated Endothelium

  • Kwon Hyang-Mi;Lim Soon Sung;Choi Yean-Jung;Jeong Yu-Jin;Kang Sang-Wook;Bae Ji-Young;Kang Young-Hee
    • Nutritional Sciences
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    • v.8 no.3
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    • pp.153-158
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    • 2005
  • Numerous natural herbal compounds have been reported to inhibit adhesion and migration of leukocytes to the site of inflammation Licorice extracts, which have been widely used in traditional Chinese medicinal preparation, possess various pharmacological effects. Isoliquiritigenin, a biogenetic precursor of flavonoids with various pharmacological effects, is a natural pigment present in licorice. We attempted to explore whether licorice extracts and isoliquiritigenin mitigate monocyte adhesion to tumor necrosis factor-$\alpha$ (TNF-$\alpha$)-activated human umbilical vein endothelial cells (HUVEC). In addition, it was tested whether the inhibition of monocyte adhesion to the activated HUVEC accompanied a reduction in vascular cell adhesion molecule-l expression(VCAM-l). Dry-roasted licorice extracts in methylene chloride but not in ethanol markedly interfered with THP-l monocyte adhesion to INF-$\alpha$-activated endothelial cells. licochalcone A compound isolated from licorice extract in methylene chloride appeared to modestly inhibit the interaction of THP-l monocytes and activated endothelium. In addition, isoliquiritigenin abolished the monocyte adhesion with attenuating VCAM-l protein expression on HUVEC induced by INF-$\alpha$. These results demonstrated that non-polar components from dry-roasted licorice extracts containing licochalcone A as well as isoliquiritigenin were active in blocking monocyte adhesion to cytokine-activated endothelimn, which appeared to be mediated most likely through the inhibition of VCAM-l expression on HUVEC. Therefore, licorice may hamper initial inflammatory events on the vascular endothelium involving induction of endothelial cell adhesion molecules.