• International Journal of Oral Biology
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• 제33권4호
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• pp.149-154
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• 2008

Porphyromonas gingivalis, a major periodontal pathogen, has been implicated in the initiation and progression of periodontal disease. Endothelial dysfunction (Editor note: Aberrant and dysfunction are somewhat redundant. The authors may want to choose one or the other.) contributes to chronic periodontal inflammation. Using cDNA-representational difference analysis, we found that P.gingivalis lipopolysaccharide differentially induces a number of genes in human microvascular endothelial cells. Among these upregulated genes, we focused on intercellular adhesion molecule-1 (VCAM-1), which is crucial for leukocyte recruitment during vascular inflammation. P. gingivalis LPS significantly increased the expression of vascular cell adhesion molecule-1 (VCAM-1) as well as ICAM-1. Promoter assays revealed that the transcription of these cell adhesion molecules was mainly regulated by nuclear factor-${\kappa}B$ (NF-${\kappa}B$) in endothelial cells. Furthermore, P. gingivalis LPS significantly increased leukocyte adhesiveness to microvascular endothelial cells and to aortic endothelium. Taken together, our results demonstrate that P. gingivalis LPS activates microvascular endothelial cells through NF-${\kappa}B$-dependent expression of cell adhesion molecules.

• Journal of Microbiology
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• 제39권2호
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• pp.142-145
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• 2001

Changes in the Activities of several antioxidant enzymes in transformed human dermal microvascular endothelial Cells (HMEC-1) by infection with the obligate intracellular bacterium Orientia tsutsugamushi, the causative agent of scrub typhus, were investigated. The activities of glucose-6-phosphate dehydrogenase, catalase, and glutathione peroxidase were significantly decreased in HMEC-1 cells infected with Ο. tsutsugamushi. However, the level of superoxide dismutase increased slightly. Furthermore, Increased levels of intracellular peroxide was observed in HMEC-1 during infection. These results support the hypothesis that cells infected by this intracellular bacterium experience oxidant-mediated injury that may eventually contribute to cell death.

• 미생물학회지
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• 제41권3호
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• pp.232-235
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• 2005

Scrub typhus의 원인병원체인 Orientia tsutsugamushi에 감염된 인간 피부 미세혈관 내피세포(HMEC-1)에서 superoxide dismutase 유전자의 발현이 증가하였다. O. tsutsugamushi 접촉 후 5 시간 이내에 감염된 세포에서 세포내 peroxide 존재를 유세포 분석을 통하여 관찰할 수 있었다. 또한 감염된 세포는 세포 내 glutathione이 심하게 고갈됨을 관찰하였다. 이 결과들은 이 절대세포내 기생세균에 감염된 숙주세포가 산화-매개 손상 받음을 시사한다.

• International Journal of Oral Biology
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• 제37권3호
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• pp.130-136
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• 2012

The GroEL heat-shock protein from Fusobacterium nucleatum, a periodontopathogen, activates risk factors for atherosclerosis in human microvascular endothelial cells (HMEC-1) and ApoE-/- mice. In this study, we analyzed the signaling pathways by which F. nucleatum GroEL induces the proinflammatory factors in HMEC-1 cells known to be risk factors associated with the development of atherosclerosis and identified the cellular receptor used by GroEL. The MAPK and NF-${\kappa}B$ signaling pathways were found to be activated by GroEL to induce the expression of interleukin-8 (IL-8), monocyte chemoattractant protein 1 (MCP-1), intercellular adhesion molecule 1 (ICAM-1), vascular cell adhesion molecule 1 (VCAM-1), E-selectin, and tissue factor (TF). These effects were inhibited by a TLR4 knockdown. Our results thus indicate that TLR4 is a key receptor that mediates the interaction of F. nucleatum GroEL with HMEC-1 cells and subsequently induces an inflammatory response via the MAPK and NF-${\kappa}B$ pathways.

• Biomolecules & Therapeutics
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• 제20권3호
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• pp.293-298
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• 2012

The purpose of this study was to investigate the modification of expression and functionality of the drug transporter P-glycoprotein (P-gp) by tumor necrosis factor-alpha (TNF-${\alpha}$) and interferon-gamma (IFN-${\gamma}$) at the blood-brain barrier (BBB). We used immortalized human brain microvessel endothelial cells (iHBMEC) and primary human brain microvessel endothelial cells (pHBMEC) as in vitro BBB model. To investigate the change of p-gp expression, we carried out real time PCR analysis and Western blotting. To test the change of p-gp activity, we performed rhodamin123 (Rh123) accumulation study in the cells. In results of real time PCR analysis, the P-gp mRNA expression was increased by TNF-${\alpha}$ or IFN-${\gamma}$ treatment for 24 hr in both cell types. However, 48 hr treatment of TNF-${\alpha}$ or IFN-${\gamma}$ did not affect P-gp mRNA expression. In addition, co-treatment of TNF-${\alpha}$ and IFN-${\gamma}$ markedly increased the P-gp mRNA expression in both cells. TNF-${\alpha}$ or IFN-${\gamma}$ did not influence P-gp protein expression whatever the concentration of cytokines or duration of treatment in both cells. However, P-gp expression was increased after treatments of both cytokines together in iHBMEC cells only compared with untreated control. Furthermore, in both cell lines, TNF-${\alpha}$ or IFN-${\gamma}$ induced significant decrease of P-gp activity for 24 hr treatment. And, both cytokines combination treatment also decreased significantly P-gp activity. These results suggest that P-gp expression and function at the BBB is modulated by TNF-${\alpha}$ or/and IFN-${\gamma}$. Therefore, the distribution of P-gp depending drugs in the central nervous system can be modulated by neurological inflammatory diseases.

• Journal of Yeungnam Medical Science
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• 제27권2호
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• pp.91-97
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• 2010

암세포의 전이, 죽상경화증, 당뇨성 망막병증과 같은 병적인 과정에서 혈관내피세포는 핵심적인 역할을 담당한다. 죽상경화증의 죽종 형성에 혈관민무늬근육세포가 직접적으로 관여한다. 배꼽정맥, 혈관내벽, 그리고 망막에 있는 이들 내피세포들은 다양한 효소용액들을 이용하여 얻는다. 순수하게 분리된 이들 세포는 내피세포와 관련된 질병의 시험관 내 연구에 있어 중요한 모델이다. 이러한 관점에서 볼 때 대동맥 벽의 중간막에서 분리한 후 배양한 민무늬근육세포도 죽상경화증의 발병을 설명할 수 있다. 이 종설에서는 사람배꼽정맥내피세포(HUVEC),대동맥의 내피세포 및 민무늬근육세포, 그리고 망막미세혈관내피세포(RMEC)의 분리 뿐 만 아니라 이들 세포를 이용한 질병연구에 관한 논문들을 소개하고자 한다.

• 대한의용생체공학회:의공학회지
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• 제16권1호
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• pp.1-8
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• 1995

A variety of experiments of endothelial cell seeding onto artificial vessels have been performed. To improve endothelialization, one or two components of the extracellular matrix (ECM) have been used as an underlying matrix. In this study, the whole ECM excreted from fibroblasts was used as an underlying matrix. Fetal human fibroblasts were cultured on a polyurethane (PU) sheet. After a conflu; ence was attained, the cytoskeleton and the nuclei of the fibroblast were destroyed using Triton-X. Mitomycin, or irradiation. Omental microvascular endothelial cells from adult human were seeded onto various substrates. After 12 days in culture, the cells were counted. It was observed that the ECM treated by irradiation had the highest cell number. In addition, the cells on this substrate exhibited the most typical endothelial cell morphology. For preliminary animal experiments the PU vessels (inner diameter, 1.5mm) coated with ECM were implanted in the infrarena] abdominal aorta of rat. After the vessels had been implanted for 5 weeks, it was found that the surface of the PU vessels was completely covered with endothelia] cells. In conclusion, we can state that the fibroblast-derived whole ECM makes a better underlying substrate for the endothelialization of small diameter artificial vessels.

• 한국고분자학회:학술대회논문집
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• 한국고분자학회 2006년도 IUPAC International Symposium on Advanced Polymers for Emerging Technologies
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• pp.46-47
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• 2006

Vascularization is essential for success in regenerative medicine. We have developed in vitro models to study how human microvascular endothelial cells (EC) and endothelial progenitor cells (EPC) colonize polymer scaffolds and express the endothelial phenotype, including angiogenesis. Examples are given of supportive growth and differeniation of EC on microfibre meshes of the silk protein fibroin and blends of starch with poly(epsilon-caprolactone), phenotypic markers being studied at both protein and mRNA level. Experimental models are also shown and concepts discussed to investigate how the stem cell niche, including that responsible for vascularization could be targeted, for example, by using engineered biodegradable polymer nanoparticles.

• 생명과학회지
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• 제26권12호
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• pp.1367-1375
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• 2016

Cilostazol은 phosphodiesterase III의 선택적 저해제로 알려져 있으며, 뇌졸중 치료에 일반적으로 사용되고 있다. Cilostazol을 처리한 경우, 국소 뇌허혈이 발생한 후에 혈관신생을 통해서 혈관형성이 향상된다는 것을 본 연구자들이 발표하였다. 혈관신생은 조직의 허혈상태를 극복하기 위해서 혈관재생을 촉진하는 중요한 과정으로써, 혈관내피세포의 증식, 이동, 모세관구조 형성의 다단계 과정으로 구성되어 있다. 이에 본 연구에서는 인간 뇌혈관내피세포를 이용하여 cilostazol이 혈관신생의 각 단계들에 어떤 영향을 미치는지 조사하였다. Cilostazol은 농도의존적으로 뇌혈관내피세포의 이동성을 촉진하였으나, 뇌혈관내피세포의 증식과 모세관구조 형성에는 영향을 미치지 않았다. Cilostazol이 세포이동을 조절하는 기전을 분석하기 위해서 cDNA microarray를 수행하였고, 세포이동에 관련성이 있는 5종의 후보 유전자들을 선택하여 real-time PCR을 통해 해당 유전자의 발현을 검증하였다. Cilostazol에 의해서 발현양이 조절되는 유전자들로써, phosphoserine aminotransferase 1 (PSAT1)와 CCAAT/enhancer binding protein ${\beta}$ ($C/EBP{\beta}$)은 발현이 증가하였고, tissue factor pathway inhibitor 2 (TFPI2), retinoic acid receptor responder 1 (RARRES1), RARRES3는 발현이 감소하였다. 이상의 결과를 통해서 cilostazol이 혈관내피세포의 이동을 촉진하여 혈관신생을 향상시킬 수 있음을 제안할 수 있으며, 뇌혈관내피세포에 대한 cilostazol의 조절기전에 대해서 더욱 상세히 규명을 한다면 혈관형성을 통하여 허혈성 질환을 치료할 수 있는 유용한 정보가 될 것으로 기대한다.

• The Korean Journal of Physiology and Pharmacology
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• 제20권4호
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• pp.325-332
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• 2016

Resveratrol, a phytoalexin, is reported to activate AMP-activated protein kinase (AMPK) in vascular cells. The blood-brain barrier (BBB), formed by specialized brain endothelial cells that are interconnected by tight junctions, strictly regulates paracellular permeability to maintain an optimal extracellular environment for brain homeostasis. The aim of this study was to elucidate the effects of resveratrol and the role of AMPK in BBB dysfunction induced by lipopolysaccharide (LPS). Exposure of human brain microvascular endothelial cells (HBMECs) to LPS ($1{\mu}g/ml$) for 4 to 24 hours week dramatically increased the permeability of the BBB in parallel with lowered expression levels of occluding and claudin-5, which are essential to maintain tight junctions in HBMECs. In addition, LPS significantly increased the reactive oxygen species (ROS) productions. All effects induced by LPS in HBVMCs were reversed by adenoviral overexpression of superoxide dismutase, inhibition of NAD(P) H oxidase by apocynin or gain-function of AMPK by adenoviral overexpression of constitutively active mutant (AMPK-CA) or by resveratrol. Finally, upregulation of AMPK by either AMPK-CA or resveratrol abolished the levels of LPS-enhanced NAD(P)H oxidase subunits protein expressions. We conclude that AMPK activation by resveratrol improves the integrity of the BBB disrupted by LPS through suppressing the induction of NAD(P)H oxidase-derived ROS in HBMECs.