• Toxicological Research
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• v.11 no.2
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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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• v.11 no.2
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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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• v.29
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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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• v.12 no.5
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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.

• Journal of Physiology & Pathology in Korean Medicine
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• v.18 no.2
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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.

• Proceedings of the PSK Conference
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• 2002.04a
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• pp.154.2-154.2
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• 2002

• BMB Reports
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• v.28 no.4
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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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• v.18 no.2
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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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• v.16 no.3
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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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• v.30 no.6
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• pp.553-558
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• 1997

Arterial allografts have known advantages over prosthetic vascular conduit for treatment of heart valvular disease, congenital heart disease and aortic disease. Cell viability may play a role in determining the longterm outcome of allografts. Endothelial cell is one important part in determining the allograft viability. To evaluate the viability of endothelial cells using current allograft preservation technique, porcine heart valve leaflets and arterial wall were subjected to collagenase digestion. Single endothelial cell suspension was labeled with GSA-PITC(Griffonia simplicifolia agglutininfluorescein isothiocyan te), a vascular, endothelial cell specific marker. The cell suspension was washed and incubated with Pl(Propidium iodide), which does not bind with viable cells, Endothelial cell viability was evaluated by calculating the percentage of GSA-FITC(+) and Pl(-) group using flowcytometric analysis. Allografts were treated with $4^{\circ}C$ antibiotic solo!ion for 24 hours for sterilization. After this, half of allografts were stored in $4^{\circ}C$ RPMI 1640 with HEPES buffer culture medium with 10% fetal bovine serum for 1 to 14 days(Group I). Another half of allografts were cryopreserved with a currently used technique (Group II). During the procurement and sterilization of arterial allografts, 22.8% and 24.4% of endothelial cell viability declined, respectively. In Group I, 11.9% of endothelial cell viability declined further steadily during 14 days of storage. In Group II, 13.7% of endothelial cell viability declined. These results show that largest loss of endothelial cell viability occurs during the nitial process. After 14 days of arterial allograft storage under $4^{\circ}C$ nutrient medium or cryopreservation, about 40% of endothelial cell viability is maintained. There were no differences between the endothelial cell viability from aortic valve leaflet, pulmonic valve leaflets, aortic wall and pulmonic wall.