• Journal of Photoscience
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• 제9권2호
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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.

• 대한한방내과학회지
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• 제39권1호
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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.

• 대한한방내과학회지
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• 제34권1호
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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.

• Biotechnology and Bioprocess Engineering:BBE
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• 제5권2호
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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.

• 생명과학회지
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• 제14권5호
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• pp.874-881
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• 2004

혈관 신생은 암의 성장 및 전이뿐만 아니라 염증, 관절염, 건성, 동맥경화 등의 병적인 진행에 주요한 역할을 하며, 혈관신생 억제를 통한 암의 치료를 시도하는 연구들이 활발하게 진행되고 있다. 혈관 신생 시 내피세포의 증식, 이동을 유도하는 활성화 과정이 필수적으로 일어나는 것으로 알려져 있다 본 연구에서는 in vitro에서 내피세포를 배양하여, 각종 growth factor가 풍부한 배지에서 활성화 시켰을 때, 그렇지 않는 세포들과의 유전자 발현 형태를 비교 조사하였다. HUVEC을 70∼80% cofluency로 배양시킨 후에 endothelial cell growth supplement (ECCS), 20% fetal bovine serum, heparin이 첨가된 Ml99 배지에서 13 시간 활성화시킨 세포(AHUVEC)와 대조군 세포(RHUVEC)로부터 분리한 total RNA로부터 CDNA를 제작하였고, 이것을 18,864 개의 유전자가 올려져있는 인간 올리고 칩과 hybridization 반응을 시켰다. 반응된 유전자를 이용하여 random clustering분석을 실시한 결과, 활성화 시켰던 HUVEC과 그렇지 않은 HUVEC으로 dendrogram 상에서 두개의 subgroup으로 나뉘어 지는 것을 확인할 수 있었다. 최소 2배 이상 발현 변화가 있는 유전자 122종이 활성화 시켰던 HUVEC으로부터 추출되었다. 이중에서 기능이 알려진 32 개의 유전자는 활성화시킨 HUVEC에서 발현이 증가하였고, 38 개의 유전자 발현은 감소하였다. 흥미롭게도 세포 증식과 이동, 염증, 면역반응에 관련한 유전자의 발현이 증가된 반면에 세포 흡착과 혈관 조직과 기능에 관련한 유전자의 발현이 감소된 것이 관찰되었다. 예상외로 규명이 잘된 혈관신생 인자와 관련한 유전자들의 발현에는 크기 차이를 보이지 않았으나, Eph-B4의 발현은 약 4 배 감소된 것으로 관찰되었다 또한, 2배 이상 발현에 차이를 보이고 기능이 알려져 있지 않은 유전자 52종이 발견되었다. 따라서, 이러한 연구 결과로부터 새로운 혈관 표적 물질 개발에 대한 기회가 제공될 수 있을 것이라 사료된다.

• 한국식품영양과학회지
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• 제40권5호
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• pp.635-641
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• 2011

본 연구는 TNF-${\alpha}$로 자극된 HUVEC에서 녹차씨껍질 EtOAC 추출물이 초기 동맥경화 과정에 중요한 역할을 하는 염증매개인자와 세포부착물질에 미치는 영향을 분석하였다. 녹차씨껍질 EtOAC 추출물의 NO 생성능은 TNF-${\alpha}$만을 처리한 control군에 비해 증가시키는 것을 알 수 있었다. $100{\mu}g$/mL 농도에서는 녹차씨껍질 EtOAC 추출물은 세포독성을 보이지 않았으며 염증매개인자인 TNF-${\alpha}$ 수준 및 세포부착물질인 VCAM-1과 MCP-1의 생성을 억제하였다. 뿐만아니라, 녹차씨껍질 EtOAC 추출물은 총 항산화능은 증가되는 경향을 보였다. 이상의 결과에서, 녹차씨껍질 EtOAC 추출물은 HUVEC에서 TNF-${\alpha}$로 인한 총 항산화능의 수준을 향상시켜 염증생성인자인 TNF-${\alpha}$ 수준 및 세포부착물질인 VCAM-1과 MCP-1의 생성을 억제하여 동맥경화 초기반응을 억제하는데 기여할 것으로 사료된다.

• 대한한의학회지
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• 제25권2호
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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.

• Archives of Pharmacal Research
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• 제28권1호
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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.

• 한국식품영양과학회지
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• 제40권3호
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• pp.385-392
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• 2011

Glyceraldehyde를 이용하여 제조된 AGEs 화합물(glycer-AGEs)을 단구세포인 THP-1, 혈관내피세포인 HUVEC 및 이 두 세포가 동시에 배양된 system에서 100 ${\mu}g/mL$로 처리 후 24시간까지 처리시간을 달리하여 처리하였다. 배양시간 동안 각 세포와 배양액을 회수하여 TNF-$\alpha$와 IL-1$\beta$의 발현을 mRNA 수준에서 조사하였다. 그 결과, THP-1의 경우 배양 2시간에서 TNF-$\alpha$나 IL-1$\beta$의 mRNA 발현이 대조구에 비해 증가되었으나 혈관내피세포인 HUVEC의 경우에는 24시간 배양하는 동안 유의적인 차이가 없었다. 그러나 두세포를 동시에 배양한 system에서 혈관내피세포인 HUVEC의 경우에는 배양 4시간에서 대조구보다 TNF-$\alpha$의 발현은 4.4배, IL-1$\beta$의 경우 5.5배 정도 증가되는 것을 확인할 수 있었다. TNF-$\alpha$와 IL-1$\beta$의 배지 내에서의 농도를 측정해 본 결과, THP-1만 배양한 경우 대조구의 배지 내 TNF-$\alpha$ 함량이 배양 6시간에 98.2 pg/mL로 대조구 53.8 pg/mL보다 증가하였으며 HUVEC의 경우 배양 8시간에 93.3 pg/mL로 증가하였다. 그러나 co-culture의 경우 배양 4시간부터 증가하여 배양 8시간에 199.2 pg/mL로 증가하였다. RAGE는 TNF-$\alpha$와 IL-1$\beta$의 발현 pattern과 다르게 단독 및 co-culture에서 배양 16시간에 대조구에 비해 각각 1.6배, 24시간에 1.9배 증가하였다. 따라서 본 연구 결과 최종당화산물에 의해 혈관내피세포의 기능상실의 연구에 있어 co-culture조건이 유용하며, 특히 mRNA 수준에서는 4시간에, protein 수준에서는 8시간에 효능을 측정하면 유효성 있는 연구 성과를 얻을 있을 것으로 예측할 수 있었다.

• Nutritional Sciences
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• 제8권3호
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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.