• 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.

• 대한한의학회지
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• 제41권2호
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• pp.43-57
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• 2020

Objectives: The purpose of this study was to investigate the effect of Phellinus linteus cheonghyeol plus (PLCP) on antioxidant and inhibition of inflammatory factor expression associated with dyslipidemia in HUVEC. Methods: The scavenging activity of DPPH and ABTS radical of PLCP was measured in HUVEC. The expression levels of NF-κB, p-IκBα, ERK, JNK, and p38 proteins were measured after treating with TNF-α in HUVEC. The expression levels of MCP-1, ICAM-1, and VCAM-1 mRNA and biomarkers were measured after treatment with TNF-α in HUVEC Results: 1. PLCP increases DPPH and ABTS radical scavenging activity in a concentration dependent manner. 2. PLCP significantly decreased the concentration of NF-κB, p-IκBα, ERK, JNK protein compared to the control at concentrations of 100 ㎍/㎖ or more, and significantly decreased concentration of p38 protein at all concentrations. 3. PLCP significantly decreased MCP-1 mRNA expression levels at 100㎍/㎖ or more compared to the control. ICAM-1 and VCAM-1 mRNA expression levels were significantly reduced at all concentrations compared to the control. MCP-1, ICAM-1 protein expression levels were significantly reduced compared to the control at concentrations of 100 ㎍/㎖ or more, and VCAM-1 protein expression levels were reduced at all concentrations. Conclusions: These results suggest that PLCP has an antioxidant effect, and it has been experimentally confirmed that it can prevent or inhibit inflammatory diseases caused by dyslipidemia due to its inhibitory effect on inflammation-related factors in HUVEC.

• 미생물학회지
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• 제50권3호
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• pp.223-226
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• 2014

청국장은 대두발효식품으로 대두 단백질이 발효 중 분해되면서 다양한 생리활성물질이 만들어진다. 혈관내피세포는 혈관의 기능은 물론 신생혈관 생성을 주도하는 세포이다. 뇌졸중이나 심근경색, 뇌경색 등의 혈관관련 질병들은 신생혈관 생성을 촉진하는 치료법이 필요하다. Vascular Endothelial Growth Factor (VEGF)는 새로운 혈관 형성을 촉진하는 역할을 한다. 본 연구에서는 청국장 에탄올추출물(CEE)이 HUVEC (혈관 내피세포) 증식에 미치는 영향을 조사하여 보았다. 청국장 추출물(100, $1000{\mu}g/ml$)을 HUVEC에 처리했을 때, VEGF (10 ng/ml)를 처리한 대조군과 같은 정도로 세포를 증식시켰다. 열처리한 청국장추출물을 혈관 내피세포에 처리해도 세포 증식효과는 마찬가지였다. 청국장이 세포 증식뿐 아니라 HUVEC이동에도 영향을 주는지 sprout 분석법으로 확인하였다. 청국장 추출물($100{\mu}g/ml$)을 처리했을 때, VEGF (10 ng/ml)와 비슷할 정도로 HUVEC 이동이 일어났다. 청국장 추출물에서 HUVEC 증식과 이동에 영향을 미치는 특정 peptide의 분리가 필요할 것이다.

• 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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• 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.

• 동의생리병리학회지
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• 제34권3호
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• pp.126-135
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• 2020

The purpose of this study was to investigate the antioxidant, anti-inflammatory and anti-cellular adhesion molecules effects of Allii Macrostemonis Bulbus, Artemisiae Capillaris Herba, Curcumae Radix, Crataegi Fructus, Salviae Militiorrhizae Radix complex extract(AMCP) on the inhibition of atherosclerosis in HUVEC. We measured DPPH radical scavenging activity and ABTS radical scavenging activity of AMCP to evaluate its antioxidant effect. And we also measured the expression level of NF-κB, IκBα, ERK, JNK, p38 proteins to evaluate its anti-inflammatory effect. Lastly, we measured the expression level of MCP-1, ICAM-1, VCAM-1 mRNA and their level to evaluate its anti-celluar adhesion molecules. AMCP did not show any cytotoxicity in HUVEC within the concentraion tested except for a concentration of 400 ㎍/㎖. AMCP increased the DPPH radical scavenging activitiy and ABTS radical scavenging activity in HUVEC as the concentration of AMCP rises. AMCP significantly reduced NF-κB, IκBα, JNK, ERK and p38 protein expression in HUVEC compared to control group. AMCP significantly reduced MCP-1, ICAM-1, VCAM-1 gene expresion in HUVEC compared to control group. AMCP significantly decreased the levels of MCP-1, ICAM-1, VCAM-1 in HUVEC compared to control group. These results suggest that AMCP has effects on antioxidation, anti-inflammation and anti-cellular adhesion molecule, which helps the treatment and prevention of dyslipidemia and atherosclerosis.

• 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.

• 대한한방내과학회지
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• 제39권4호
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• pp.480-494
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• 2018

Objectives: To investigate the effect of fermented extract of Artemisiae Iwayomogii Herba, Curcumae Longae, Crataegi Fructus and Salviae Miltiorrhizae Radix (FMH) on anti-inflammation associated with dyslipidemia and anti-oxidation in RAW264.7 and HUVEC cells. Methods: The total polyphenols, total flavonoids, DPPH radical scavenging activity, ABTS radical scavenging activity, and cytotoxicity of FMH were measured. RAW264.7 cells treated with FMH were tested for production of NO, and for cytokine and LTB4 levels and HUVEC cells treated with FMH were examined for production of cDNA of genes related to inflammation. Results: 1. FMH contained polyphenols and flavonoids. The DPPH and ABTS radical scavenging activity of FMH increased in a concentration-dependent manner. 2. FMH treatment inhibited the production of nitric oxide (NO), cytokines, and LTB4 in RAW264.7 cell when compared to the untreated control group. 3. FMH decreased the transcription of pro-inflammatory genes, whereas it increased transcription of anti-inflammatory genes, in HUVEC cells. Conclusion: FMH is effective as an antioxidant and for treatment and prevention of dyslipidemia, atherosclerosis, ischemic heart disease, stroke, and other cardiocerebrovascular diseases.

• 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.