• Toxicological Research
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• 제11권2호
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• pp.169-173
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• 1995

The effects of tumor necrosis factor alpha $(TNF-{\alpha})$ on growth and tubular formation of bovine aortic endothelial cells were examined using an in vitro angiogenesis model system. The growth of endothelial cells was enhanced in a dose-dependent manner when the cells were cultured with $TNF-{\alpha}$ for 3 days, but $TNF-{\alpha}$, at the concentration of 1 nM or higher, produced a growth inhibition of endothelial cells when the cells were cultured for 8 days. The endothelial cells incubated with $TNF-{\alpha}$ for 48-h exhibited a typical morphologic change. Then, they showed a fibroblastoid organization of overlapping, elongated, and spindle-shaped cells. $TNF-{\alpha}$, at the concentration of O. 1 nM or higher, inhibited the tubular formation of vascular endothelial cells in an in vitro anglogenesis model using a 3-dimensional culture system.

• Toxicological Research
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• 제11권2호
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• pp.175-180
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• 1995

We found that bufalln, one of the prominent components of the bufadlenolides in the Chinese medicine chan'su, has the potent inhibitory effects on growth and proliferation of the cultured bovine aortlc endothelial (BAE) and human umbilical vein endothelial (HUVE) cells. All naturally-occuring bufadienolides used in this study inhibited the cell growth in a dose-dependent manner. Particularly, bufalin among the bufadienolides showed the strongest inhibitory activity for the cell growth. The order of growth inhibition by bufadienolides on BAE cells was as follows: bufalin > gamabufotalln > bufotalln > cinobufagin > cinobufotalin > resibufogenin. The $IC_50$ values (50% inhibition of cell growth) of bufalin as determined by XTT assay were the range of 1-10 nM in BAE and HUVE cells. Bufalin exhibited a higher sensitivity towards cultured bovine aortic endothelial cells than human umbilical vein endothelial cells.

• Environmental Analysis Health and Toxicology
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• 제29권
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• pp.10.1-10.6
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• 2014

Objectives Cigarette smoking had been recorded as the main cause of impaired endothelium-dependent vasodilation in smokers by reducing nitric oxide (NO), a production of endothelial nitric oxide synthase (eNOS). However, the mechanism of NO impairment via eNOS activity is unclear until now. In this study, cell passage is suggested to be a relevant factor to eNOS expression under cigarette smoking stress. Methods Bovine aortic endothelial cells (BAECs) were chosen as the research subject with passages ranking from 6 to 9 (6P to 9P). After exposure of cigarette smoking extract (CSE) solution, MTT assay and Western blot method were performed to check the cell viability as well as eNOS protein concentration. In these experiments, four concentrations of CSE at 0.5, 1, 2, and 4% were selected for treatment. Results Our results showed that cells almost died at 4% of CSE. Besides, eNOS protein mass had a linear decrease under the increase of CSE concentration. In addition, the effect of CSE on eNOS expression was dissimilar between different passages. Conclusions This study indicated that CSE had effect on both cell viability and eNOS expression. Besides, a reduction in protein mass was matched with the decrease of cell viability due to CSE tress. Last but not least, the response of eNOS protein to different concentration of CSE at different passages was disparate, making the hypothesis about cell passage related inhibition of eNOS caused by CSE solution.

• Advances in nano research
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• 제12권5호
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• pp.501-514
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• 2022

This study aimed to investigate the effects and potential mechanisms of Chikusetsusaponin V (CsV) on endothelial nitric oxide synthase (eNOS) and vascular endothelial cell functions. Different concentrations of CsV were added to animal models, bovine aorta endothelial cells (BAECs) and human umbilical vein endothelial cells (HUVECs) cultured in vitro. qPCR, Western blotting (WB), and B ultrasound were performed to explore the effects of CsV on mouse endothelial cell functions, vascular stiffness and cellular eNOS mRNA, protein expression and NO release. Bioinformatics analysis, network pharmacology, molecular docking and protein mass spectrometry analysis were conducted to jointly predict the upstream transcription factors of eNOS. Furthermore, pulldown and ChIP and dual luciferase assays were employed for subsequent verification. At the presence or absence of CsV stimulation, either overexpression or knockdown of purine rich element binding protein A (PURA) was conducted, and PCR assay was employed to detect PURA and eNOS mRNA expressions, Western blot was used to detect PURA and eNOS protein expressions, cell NO release and serum NO levels. Tube formation experiment was conducted to detect the tube forming capability of HUVECs cells. The animal vasodilation function test detected the vasodilation functions. Ultrasonic detection was performed to determine the mouse aortic arch pulse wave velocity to identify aortic stiffness. CsV stimulus on bovine aortic cells revealed that CsV could upregulate eNOS protein levels in vascular endothelial cells in a concentration and time dependent manner. The expression levels of eNOS mRNA and phosphorylation sites Ser1177, Ser633 and Thr495 increased significantly after CsV stimulation. Meanwhile, CsV could also enhance the tube forming capability of HUVECs cells. Following the mice were gavaged using CsV, the eNOS protein level of mouse aortic endothelial cells was upregulated in a concentration- and time-dependent manner, and serum NO release and vasodilation ability were simultaneously elevated whereas arterial stiffness was alleviated. The pulldown, ChIP and dual luciferase assays demonstrated that PURA could bind to the eNOS promoter and facilitate the transcription of eNOS. Under the conditions of presence or absence of CsV stimulation, overexpression or knockdown of PURA indicated that the effect of CsV on vascular endothelial function and eNOS was weakened following PURA gene silence, whereas overexpression of PURA gene could enhance the effect of CsV upregulating eNOS expression. CsV could promote NO release from endothelial cells by upregulating the expression of PURA/eNOS pathway, improve endothelial cell functions, enhance vasodilation capability, and alleviate vessel stiffness. The present study plays a role in offering a theoretical basis for the development and application of CsV in vascular function improvement, and it also provides a more comprehensive understanding of the pharmacodynamics of CsV.

• 동의생리병리학회지
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• 제18권2호
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• pp.496-499
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• 2004

We have previously observed that Bambusae Caul is in Liquamen (BCL) stimulates the adipose conversion of 3T3-L1 cells and molecular chaperones were involved in the process of the assembly and replacement of laminin subunits in Bovine aortic endothelial cells(BAEC). Endothelial cells are exposed to continuous shear stress due to the blood flow. Heat shock protens(hsp) are a well-known stress response protein, namely, stress proteins. To investigate effects of BCL on the stress proteins induced by heating in endothelial cells, we have analyzed synthetic amounts of stress proteins in sodium dodecyl sulfate gel electrophoresis under reducing conditions. Under the condition of heating stress, BCL inhibited the synthesis of stress proteins in endothelial cells. These results suggest that BCL may have an important role for expression of stress proteins induced by heating in endothelial cells.

• 대한약학회:학술대회논문집
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• 대한약학회 2002년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.1
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• pp.154.2-154.2
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• 2002

• BMB Reports
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• 제28권4호
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• pp.301-305
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• 1995

Considering the stimulatory effects of ginsenosides from Panax ginseng C. A. Meyer on the release of nitric oxide from bovine aortic endothelial cells in vitro and vasodilatation of rabbit pulmonary artery in vivo, the present study is designed to investigate the mechanism of nitric oxide release by ginsenosides in calf pulmonary artery endothelial cells, Nitric oxide release was determined in endothelial cells treated with ginsenosides and compared with those of the receptor-dependent agonists, bradykinin and ADP and the receptor-independent calcium ionophore $A_{23187}$. The results showed that total saponin and ginsenoside $Rg_1$, not $Rb_1$, stimulated nitric oxide release measured as conversion to L-citrulline. The nitric oxide releasing properties of total saponin and ginsenoside $Rg_1$ were different; total saponin stimulated only conversion to L-citrulline, like $A_{23187}$, while ginsenoside $Rg_1$ stimulated both L-arginine transport and conversion to L-citrulline, as bradykinin or ADP did.

• Journal of Microbiology and Biotechnology
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• 제18권2호
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• pp.392-395
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• 2008

In an attempt to delineate the direct effect of arsenite-induced endothelial dysfunction on nitric oxide (NO) production, confluent bovine aortic endothelial cells (BAEC) were incubated with arsenite, and endothelial NO synthase expression and NO production were measured. Exposure of arsenite decreased NO production for up to 24h. This decrease was accompanied by decreases in cAMP, protein kinase A (PKA) activity, and furthermore, significant reduction of pCREB. In conclusion, this study is the first to demonstrate that exposure of arsenite decreases NO production by a reduction of pCREB and PKA activity that may be mediated by cAMP, leading to endothelial dysfunction.

• Journal of Microbiology and Biotechnology
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• 제16권3호
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• pp.475-479
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• 2006

Gentisyl alcohol isolated from Penicillium sp. has an antioxidative activity, protecting cells from oxidative stresses. From our in vitro angiogenesis assays with bovine aortic endothelial cells (BAECs), gentisyl alcohol was newly identified as a pro-angiogenic small molecule that induces new blood vessel formation of the cells. Gentisyl alcohol stimulated the proliferation of BAECs in a dose-dependent manner. Moreover, it induced in vitro angiogenesis of BAECs such as invasiveness, migration, and tube formation of the endothelial cells. Effects of gentisyl alcohol on invasion and tube formation were also dose-dependent. These results demonstrate that gentisyl alcohol could affect the angiogenic phenotypes of endothelial cells and be developed as a new small molecule with pro-angiogenic activity.

• 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.