• 대한한의학회지
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• 제25권1호
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• pp.49-59
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• 2004

Objectives: This paper is to investigate what effects Yeonryunggobondan and Palmijihwangtang have on postponing senility in rat fibroblasts, heart-endothelial cells, mesangial cells. Methods: 1. In vitro Yeonryunggobondan and Palmijihwangtang controlled the growth of fibroblasts, heart-endothelial cells, mesangial cells, extended the PDT of them. 2. After feeding rats the drugs for 2 months, the fibroblasts, heart-endothelial cells, mesangial cells were cultured. Results: 1) In fibroblasts the PDN was incresed and the PDT was decreased at passage-1, 2 by Yeonryunggobondan and Palmijihwangtang(p<0.05). 2) In heart-endothelial cells the PDN was incresed and the PDT was decreased at passage 8 by Yeonryunggobondan and Palmijihwangtang(p<0.05). 3) In mesangial cells the PDN was increased and the PDT was decreased at passage 4 by Yeonryunggobondan, the PDN was incresed at passage 4 by Palmijihwangtang(p<0.05). Conclusions: It is concluded that both Yeonryunggobondan and Palmijihwangtang maybe be conductive to protect and delay the senescence of rat fibroblasts, heart-endothelial cells, mesangial cells.

• Molecules and Cells
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• 제37권4호
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• pp.330-336
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• 2014

Thymosin beta4 (TB4) has multiple functions in cellular response in processes as diverse as embryonic organ development and the pathogeneses of disease, especially those associated with cardiac coronary vessels. However, the specific roles played by TB4 during heart valve development in vertebrates are largely unknown. Here, we identified a novel function of TB4 in endothelial-mesenchymal transformation (EMT) in cardiac valve endocardial cushions in zebrafish. The expressions of thymosin family members in developing zebrafish embryos were determined by whole mount in situ hybridization. Of the thymosin family members only zTB4 was expressed in the developing heart region. Cardiac valve development at 48 h post fertilization was defected in zebrafish TB4 (zTB4) morpholino-injected embryos (morphants). In zTB4 morphants, abnormal linear heart tube development was observed. The expressions of bone morphogenetic protein (BMP) 4, notch1b, and hyaluronic acid synthase (HAS) 2 genes were also markedly reduced in atrio-ventricular canal (AVC). Endocardial cells in the AVC region were stained with anti-Zn5 antibody reactive against Dm-grasp (an EMT marker) to observe EMT in developing cardiac valves in zTB4 morphants. EMT marker expression in valve endothelial cells was confirmed after transfection with TB4 siRNA in the presence of transforming growth factor ${\beta}$ ($TGF{\beta}$) by RT-PCR and immunofluorescent assay. Zn5-positive endocardial AVC cells were not observed in zTB4 morphants, and knockdown of TB4 suppressed TGF-${\beta}$-induced EMT in ovine valve endothelial cells. Taken together, our results demonstrate that TB4 plays a pivotal role in cardiac valve formation by increasing EMT.

• Journal of Chest Surgery
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• 제37권4호
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• pp.297-306
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• 2004

기존의 인공 심장 판막은 혈전 형성, 내구성, 세균 감염에 대한 저항력, 성장성 등에서 문제점을 가지고 있다 최근 이상적인 심장 판막을 개발하기 위한 노력으로 조직공학 기법을 이용하고 있다. 본 연구는 이종 판막을 지지체로 하여 이종 세포 제거 후 자가 세포를 파종한 조직공학 심장 판막을 개발하기 위하여 이종 세포 제거와 자가 세포 파종 방법을 비교하였고, 얻어진 심장 판막첨의 생체 적합성을 살펴보았다. 대상 및 방법: 이종 지지체로 두 마리의 돼지에서 각각 3개의 폐동맥 판막첨을 채취하여, Triton-X, freeze-thawing, NaCl-SDS 등 세 방법 중 하나로, 각각 두 개의 판막첨을 처리하고, 세포 제거 정도를 비교하였다. 염소의 경정맥에서 분리한 내피 세포를 배양하여, 상기한 방법 중 하나로 이종 세포가 제거된 돼지 판막첨에 파종한 후, 내피 세포의 보존 상해를 비교하였다. 생체 실험을 위해, 6마리 돼지에서 각각 2개의 폐동맥 판막첨을 채취해 이종 세포를 제거하였고, 6개의 판막첨은 염소의 내피 세포를 파종하고(이종-자가 이식편),다른 6개의 판막첨은 내피 세포를 파종않고(이종 이식편) 6마리 염소의 폐동맥 판막첨 두 개와 치환하였다. 치환되지 않은 하나의 판막첨을 대조군으로 비교하였다. 염소는 수술 6시간, 24시간, 1주일, 1개월, 3개월, 그리고 6개월 후 희생시키고 폐동맥 판막첨을 채취하여 분석하였다 걸과: Triton-X와 freeze-thawing 처리한 판막첨에서는 이종세포가 남아 있었으나, NaCl-SDS 처리한 판막첨에서는 세포가 완전히 제거되었고, 이종 세포 제거 후 파종한 자가 내피 세포도 NaCl-SDS 처리한 판막첨에서 다른 두 방법으로 처리한 판막첨에 비해 잘 보존되어 있었다. 6마리 염소는 혈색전증의 소견 없이 예정된 기간 동안 모두 생존하였으며, 희생 후 적출한 이종 이식편과 이종-자가 이식편 모두 대조군 보다 두꺼워져 있었다. 두 이식편 모두 24시간 후부터 섬유아세포의 증식이 관찰되었고, 1개월 후 세포의 개형(remodeling)과정을 볼 수 있었고, 3개월 후 세포 기능을 수행하였고, 6개월 후 대조군보다 세포수가 더 많이 증가하였다. 파종한 내피 세포는 잘 보존되어 있었고, 이종 이식편도 6개월 후에는 내피 세포가 피복 되어 있었다. 걸론: 이종 세포의 제거와 자가 세포의 파종에 가장 적합한 방법은 NaCl-SDS 처리였다 동물 실험에서 이종 이식편과 이종-자가 이식편 모두 6개월 간 지지체의 역할을 수행하면서, 자가 세포의 증식을 가능하게 하였고, 자가 조직으로 발전할 가능성을 보여 주었다.

• Journal of Nutrition and Health
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• 제31권8호
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• pp.1235-1243
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• 1998

Although an elevated plasma level of high density lipoprotein (HDL) is known as a protective component against the development of atherosclerosis and ensuing coronary heart diseases, the related mechanisms are still not established . It has been clearly demonstrated in the early stages of atherogenesis that adhesion of monocytes and lymphocytes to the vascular endothelium is enhanced via adhesion molecules, and that monocytes and macrophages accumulate in the subendothelial space. The present study has investigated whether isolated plasma HDL plays a role in protection against atherogenesis by inhibiting the expression of vascular cell adhesioin molecule-1(VCAM-1) on the endothelial cells. Effects of plasma native low density lipoprotein (LDL) and ac ethylated LDL(AcLDL) on VCAM-1 expression were also examined by using an immunocytochemical technique. While plasma HDL did not alter the basal expression of VCAM-1 , lipopolysaccharide(LPS) induction of this adhesion modlecule was markedly inhibited at a phyaiological concentration of HDL. In contrast, 30$\mu\textrm{g}$ protein/ml AcLDL increased sifnificantly both basal VCAM-1 expression and its LPD induction , suggesting that this modified LDL enhances leukocyte adhesiion to endothelial cells. Unlike AcLDL , plasma native LDL inhibited significantly VCAM-1 expression. This indicates that LDL did not undergo oxidative modificantion while incubated with endothelial cells. These results suggest that plasam HDL may inhibit atherogenesis by reducing the expression of adhesion molecules, which is a protective mechanism independent of tis reverse cholesterol transport function . Modified LDL is a potent iducer for adhesion molecules in vascular endothelical cells and could play a role in the pathogenesis of atherosclerosis by adhering to blood cells.

• Journal of Chest Surgery
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• 제30권6호
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• pp.553-558
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• 1997

동종동맥판은 심장판막질환, 선천성 심기형 및 대동맥 질환의 치료에 있어서 우수한 판막도관으로 사용 되고 있다. 이 때 동종동맥 판의 장기성적을 좌우하는데 있어서 혈관내피세포의 생육성이 중요한 역활을 할 것이다. 혈관내피세포의 생육성을 평가하기 위하여 현재 임상에서 사용되는 보존방법으로 보존처리된 성돈의 대동맥판 및 대동맥 벽을 collagenase로 분해시켜서 순수한 내퍼세포군을 획득한 뒤, 혈관내피세포에 특이한 친화성을 갖는 GSA-FTTC(Criffonia simplicifolia agglutininfluorescein isothiocyanate)와 반응시켰다. 이 내피세포군을 세척한 다음, 살아있는 세포에는 침착되지 않는 Pl(Ropidium iodide)와 반응시켰다. 이렇게 처리된 내피세포군을 Row Cytometry 로 분석하여 GSA-FTIC(+), Pl(-) 인 세포를 생육성을 유지한 것으로 평가하였다. 동종동맥판은 $4^{\circ}C의$ 멸균용액에 24시간 담궈 멸균처리를 한 후, 2개군으로 나누어 (1군)은 $4^{\circ}C$ RPM 1640 with HEPES buffer cultlue medium with 10% fetal bovine uTm 용액에 1~14일간 보존하였고 (2군)은 냉동보존을 하였다. 조직의 획득과정과 멸균과정에서 각각 22.8%와 24.4%의 생육성이 소\ulcorner되었다. (1군) 에서는 14일의 보존기간 동안 11.9%의 생육성감소가 일어났고 (2군) 에서는 13.7%의 생육성감소가 일어났다. 이 실험의 결과로 동종동맥 판의 보존처리과정 초기에 대부분의 생육성소실이 일어나며, 14일간의 냉장보존이나 냉동보존 후에도 약 40%의 생육성이 보존됨을 알 수 있었다. 또한 혈관내피세포가 판막에서 얻어진 경우나 동맥벽에서 얻어진 경우에서 생육성의 차이는 없었다.

• 대한미생물학회지
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• 제35권3호
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• pp.203-214
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• 2000

Porphyromonas gingivalis has been implicated in periodontal diseases. Accumulating evidence suggests that cardiovascular disease is the most prevalent medical problem in patients with periodontal diseases. In order to check the possibility that P. gingivalis is involved in coronary heart disease, the present study was performed to observe P. gingivalis adherence and invasion of human coronary artery endothelial cells (HCAEC) and production of cytokines and growth factors by HCAEC upon P. gingivalis infection. $^3H$-labeled P. gingivalis 381 was incubated with HCAEC for 90 min. The radioactivity of the washed HCAEC was a measure of the absorbed (adhering and invading) P. gingivalis. The absorption radioactivity of the HCAEC infected by P. gingivalis was determined to be 59.58% of the input bacterial cells. In contrast, the absorption radioactivity of the cells infected by S. gordonii Challis which was employed as a control was negligible (0.59%). DPG3, a P. gingivalis mutant defective of fimbriae, appeared to be impaired to some extent in capability of adherence/invasion as compared to that of the parental strain 381, showing 43.04% of the absorption radioactivity. The absorption radioactivity of the HCAEC infected by P. gingivalis 381 in the presence of excessive fimbriae at the concentrations of $50\;{\mu}g$ and $100\;{\mu}g/ml$ was 57.27 and 45.44%, respectively. Invasion of HCAEC by P. gingivalis 381 was observed by an antibiotic (metronidazole) protection assay and transmission electron microscopy (TEM). In the antibiotic protection assay, invasion by the bacterium was measured to be 0.73, 1.09, and 1.51% of the input bacterial cells after incubation for 30, 60, and 90 min, respectively. Invasion by DPG3 was shown to be 0.16% after 90-min incubation. In comparison of invasion efficiency at 90 min of the incubation, the invasion efficiency of DPG3 was 0.37% while that of its parental strain 381 was 2.54%. The immunoblot analysis revealed fimbriae of P. gingivalis did not interact with the surface of HCAEC. These results suggest that fimbriae are not the major contribution to the adherence of P. gingivalis to HCAEC but may be important in the invasion of HCAEC by the bacterium. The presence of cytochalasin D ($1\;{\mu}g/ml$) and staurosporine ($1\;{\mu}M$) reduced the invasion of HCAEC by P. gingivalis 381 by 78.86 and 53.76%, respectively, indicating that cytoskeletal rearrangement and protein kinase of HCAEC are essential for the invasion. Infection of P. gingivalis induced HCAEC to increase the production of TNF-${\alpha}$. by 60.6%. At 90 min of the incubation, the HCAEC infected with P. gingivalis cells was apparently atypical in the shape, showing loss of the nuclear membrane and subcellular organelles. The overall results suggest that P. gingivalis may cause coronary heart disease by adhering to and invading endothelial cells, and subsequently damaging the cells.

• 생명과학회지
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• 제11권2호
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• pp.111-120
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• 2001

The pathological aspects of purified ricin from the seeds of the castor oil plant, Ricinus communis, were examined, using light and transmission electron microscopy. ICR mice were exposed to ricin by peritoneal injection with 100 ng/1 $m\ell$ PBS(pH 7.0) mouse and histological observations on the liver, spleen, thymus, lung and heart were carried out at intervals up to 48 h after exposures. All the organs examined were damaged by ricin. Among the organs, the spleen and thymus; immune organs were the most sensitive to ricin, whereas the effect delayed in the liver, lung and heart. Furthermore, the immune cells in each organ were the most sensitive to ricin. Accordingly, the effect of ricin on the organs seems to be affected by the immune cells existed in each organ, In each organ, the immune cells showed apoptotic changes, while the capillary endothelial and parenchymal cells showed necrotic changes.

• Environmental Analysis Health and Toxicology
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• 제21권1호
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• pp.71-79
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• 2006

Epidemiologic studies have showed a close correlation between arsenic exposure and heart disease such as, cardiovascular problem, ischemic heart disease, infarction, atherosclerosis and hypertension in human. It may increase the mortality of high risk group with heart disease. Regarding the mechanism studies of heart failure, blood vessel, vascular smooth muscle cells and endothelial cells have long been focused as the primary targets in arsenic exposure but there are only a few studies on the cardiomyocytes. In this study, the generation of oxidative stress by low dose of arsenic trioxide was investigated in rat cardiomyocytes. By direct measurement of reactive oxygen species and fluorescent microscopic observation using fluorescent dye 2',7'-dichlorofluorescin diacetate, reactive oxygen species were found to be generated without cell death, where cells are treated with 0.1 ppm arsenic for 24 hours. With the induction of reactive oxygen species, GSH level was decreased by the same treatment. However, DNA damage did not seem to be serious by DAPI staining, while high dose of arsenic (2 ppm for 24 hrs) caused fragmentation of DNA. To identify the molecular biomarkers of low-dose arsenic exposure, gene expression was also investigated with whole genome microarray. As results, 9,022 genes were up-regulated including heme oxygenase-l and glutathione S-transrerase, which are well-known biomarkers of oxidative stress. 9,404 genes were down-regulated including endothelial type gp 91-phox gene by the treatment of 0.1 ppm arsenic for 24 hours. This means that biological responses of cardiomyocytes may be altered by ROS induced by low level arsenic without cell death, and this alteration may be detected clearly by molecular biomarkers such as heme oxygenase-1.

• 한국산학기술학회논문지
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• 제18권4호
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• pp.366-379
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• 2017

본 연구는 운동중재가 심장질환자의 혈관내피전구세포에 미치는 효과에 대한 선행연구들을 체계적으로 고찰하고, 그 효과에 대한 메타분석을 위해 실시되었다. 국내외 데이터베이스인 Cochrane Library, PubMed, EMBASE, ScienceDirect, CINAHL, Scopus, KoreaMed, KISS, RISS, KMBASE 온라인 검색을 실시하였고, 검색어는 심질환, 관상동맥질환, 심부전, 심혈관질환, 운동, 신체활동, 재활, 혈관내피전구세포를 조합하여 사용하였다. 그 결과, 539편의 논문이 검색되었고, 논문 선정기준에 부합하는 9편의 논문을 최종 분석에 이용하였다. Comprehensive Meta-Analysis version 2.0을 활용하여 효과크기, 출판편중을 분석하였다. 운동군의 혈관내피성장인자(VEGF), 혈관내피세포의 수(CD34+KDR+), 혈관내피세포의 기능(FMD)은 대조군에 비해 각각 2.008 (95% CI 0.204-3.812), 1.399 (95% CI 0.310-2.489), 1.881 (95% CI 0.848-2.914) 효과크기가 나타났다. 따라서 운동 중재가 혈관내피성장인자와 혈과내피전구세포의 수를 증가시키고, 혈관내피세포의 기능을 향상시키는데 효과가 있음을 알 수 있다. 국내 심혈관질환자의 유병률과 사망률이 증가하고 있음을 고려할 때, 심혈관질환자를 대상으로 한 운동중재의 효과를 분석한 본 연구결과는 심혈관질환자의 운동중재를 계획하는데 있어 실질적인 가이드라인을 제시할 수 있을 것이다.

• Archives of Plastic Surgery
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• 제37권1호
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• pp.7-11
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• 2010

Purpose: Vascular endothelial growth factor (VEGF) is known as a growth factor of endothelium and fibroblast. The purpose is to know the VEGF effects on fibroblast proliferation and fibroblast's notch receptor expression. Methods: CCD-986sk fibroblast was purchased from the Korean Cell Bank and was used in XTT assay for proliferation and wound healing assay for migration. Immunofluorescent (IF) staining and western blotting were used in testing notch expression of fibroblast. Semiquantitative RT-PCR was used in checking notch 1 mRNA production by fibroblast. Student-t test was used for analyzing results. Results: Cell proliferation assay using XTT showed significant higher proliferation in VEGF treated fibroblast, $2.324{\pm}0.0026$ vs. $2.463{\pm}0.017$ (p=0.002). Wound healing assay showed longer migration in VEGF treated fibroblast (p=0.062). The fluorescence was brighter in VEGF treated cells of notch 1 IF staining. Notch 1 expressions and mRNA productions increased more in VEGF treated cells. Conclusion: VEGF stimulates fibroblast to proliferate, migrate and to express Notch 1 simultaneously. Notch receptor could be related to VEGF mediated wound healing.