scholarly journals Enhancement of Porphyromonas gingivalis-Lipopolysaccharide Induced MCP-1 Expression by High Glucose in Human Endothelial Cells

2020 ◽  
pp. 1-6
Author(s):  
Kazuo Sonoki ◽  
Kazuo Sonoki ◽  
Kosuke Muraoka ◽  
Hisako Hikiji
Keyword(s):  
Endothelial Cells ◽  
Vitamin C ◽  
Periodontal Disease ◽  
Mrna Expression ◽  
Porphyromonas Gingivalis ◽  
High Glucose ◽  
Synergistic Effects ◽  
Diabetic Patients ◽  
Simultaneous Stimulation ◽  
Porphyromonas Gingivalis Lipopolysaccharide

To investigate the synergistic effects of periodontal disease and diabetes mellitus on atherosclerosis, we evaluated the monocyte chemoattractant protein-1 (MCP-1) expression of endothelial cells induced by Porphyromonas gingivalis-lipopolysaccharide (LPS) and high glucose. We also tested whether antioxidants could inhibit the MCP-1 expression induced by the simultaneous stimulation of LPS and high glucose. Human umbilical vein endothelial cells (HUVECs) were stimulated with various concentrations of P. gingivalis-LPS (0.1, 1.0, and 10 µg/mL) in normal glucose (5.5 mM), with high glucose (10 mM and 20 mM), and with 0.1 µg/mL P. gingivalis-LPS in high glucose. MCP-1 expressions were measured by realtime RT-PCR and ELISA. Vitamin C (100 µM) and vitamin E (50 µM) were administered before simultaneous stimulation with 0.1 µg/mL P. gingivalis-LPS and 20 mM glucose. LPS ≥ 1.0 μg/mL increased the expression of MCP-1 mRNA and protein compared to unstimulated HUVECs. High glucose in the culture medium increased the MCP-1 mRNA expression slightly but significantly at 2 hr of incubation, but the MCP-1 protein level was not increased. Simultaneous stimulation with 0.1 μg/mL LPS and high glucose increased the expression of MCP-1 mRNA and protein compared to unstimulated HUVECs. By contrast, pre-incubation of vitamin C or E inhibited the increase of MCP-1 mRNA expression induced by 0.1 μg/mL LPS and 20 mM glucose. Our finding that high glucose enhanced the MCP-1 expression with even a low level of LPS suggests that caution is advisable regarding the atherogenicity of diabetic patients with periodontal disease.

scholarly journals Chrysin Inhibits High Glucose-Induced Migration on Chorioretinal Endothelial Cells via VEGF and VEGFR Down-Regulation

2020 ◽  
Vol 21 (15) ◽  
pp. 5541
Author(s):  
Zhen-Yu Liao ◽  
I-Chia Liang ◽  
Hsin-Ju Li ◽  
Chia-Chun Wu ◽  
Huey-Ming Lo ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Mrna Expression ◽  
Mtt Assay ◽  
High Glucose ◽  
Biological Activities ◽  
Diabetic Patients ◽  
Vegf Receptor ◽  
Expression Levels ◽  
Phosphorylated Akt ◽  
Scratch Wound

Background: Diabetes mellitus (DM) is a chronic inflammatory disease, which causes multiple complications. Diabetic retinopathy (DR) is among these complications and is a dominant cause of vision loss for diabetic patients. Numerous studies have shown that chrysin, a flavonoid, has many biological activities such as anti-oxidation and anti-inflammation. However, it is rarely used in ocular diseases. In this study, we examined the inhibitory effects of flavonoid on high glucose induced migration of chorioretinal endothelial cells (RF/6A cells) and its mechanism. Materials and methods: The viability of RF/6A cells treated with chrysin was examined with a 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay. The migration of RF/6A cells was assessed by the transwell migration and scratch wound assays. The expression of AKT, ERK, vascular endothelial growth factor (VEGF), HIF−1α and MMP-2 were determined by western blotting. To observe the mRNA expression of VEGF receptor (VEGFR), qRT-PCR, was utilized. Results: The results showed that chrysin can dose-dependently inhibit the RF/6A cell migration in vitro transwell and the scratch wound assays which are induced by high glucose. After pretreatment of RF/6A cells with different concentrations of chrysin, they did not produce any cytotoxicity in MTT assay. Moreover, chrysin down-regulated both phosphorylated AKT and ERK, as well as attenuated the expression levels of MMP-2. It also decreased the expression of the VEGF transcription factor and VEGF. Furthermore, it was shown that chrysin could suppress the protein and mRNA expression levels of VEGFR. Conclusion: The results indicate that chrysin could down-regulate the phosphorylation of AKT, ERK and MMP-2 and reduce the effects of VEGF and VEGFR in a high glucose environment. It further inhibits the high glucose-induced migration of RE/6A cells. Therefore, chrysin may have the potential for visual protection.

scholarly journals Effects of High Glucose Concentration on Pericyte-Like Differentiated Human Adipose-Derived Mesenchymal Stem Cells

2021 ◽  
Vol 22 (9) ◽  
pp. 4604
Author(s):  
Giuliana Mannino ◽  
Anna Longo ◽  
Florinda Gennuso ◽  
Carmelina Daniela Anfuso ◽  
Gabriella Lupo ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Mrna Expression ◽  
Inflammatory Cytokines ◽  
High Glucose ◽  
Angiogenic Factors ◽  
Diabetic Patients ◽  
Ros Production ◽  
Migration Ability ◽  
And Migration

A pericyte-like differentiation of human adipose-derived mesenchymal stem cells (ASCs) was tested in in vitro experiments for possible therapeutic applications in cases of diabetic retinopathy (DR) to replace irreversibly lost pericytes. For this purpose, pericyte-like ASCs were obtained after their growth in a specific pericyte medium. They were then cultured in high glucose conditions to mimic the altered microenvironment of a diabetic eye. Several parameters were monitored, especially those particularly affected by disease progression: cell proliferation, viability and migration ability; reactive oxygen species (ROS) production; inflammation-related cytokines and angiogenic factors. Overall, encouraging results were obtained. In fact, even after glucose addition, ASCs pre-cultured in the pericyte medium (pmASCs) showed high proliferation rate, viability and migration ability. A considerable increase in mRNA expression levels of the anti-inflammatory cytokines transforming growth factor-β1 (TGF-β1) and interleukin-10 (IL-10) was observed, associated with reduction in ROS production, and mRNA expression of pro-inflammatory cytokines interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α), and angiogenic factors. Finally, a pmASC-induced better organization of tube-like formation by retinal endothelial cells was observed in three-dimensional co-culture. The pericyte-like ASCs obtained in these experiments represent a valuable tool for the treatment of retinal damages occurring in diabetic patients.

ErbB2 Expression on Left Ventricular Epicardial Endothelial Cells and CD105+ Cells is Decreased in Patients with Diabetes Mellitus.

2021 ◽  
Author(s):  
Joanne T. deKay ◽  
Joshua Carver ◽  
Bailey Shevenell ◽  
Angela M. Kosta ◽  
Sergey Tsibulnikov ◽  
...  
Keyword(s):  
Diabetes Mellitus ◽  
Endothelial Cells ◽  
Cell Surface ◽  
High Glucose ◽  
Artery Bypass ◽  
Surface Expression ◽  
Left Ventricular ◽  
Cell Surface Expression ◽  
Erbb Receptors ◽  
Diabetic Patients

Abstract Background We investigated the cell surface expression of ErbB receptors on left ventricular (LV) epicardial endothelial cells and CD105+ cells obtained from cardiac biopsies of patients undergoing coronary artery bypass grafting surgery (CABG). Methods Endothelial cells and CD105+ non-endothelial cells were freshly isolated from LV epicardial biopsies obtained from 15 subjects with diabetes mellitus (DM) and 8 controls. The expression of ErbB recepotrs was examined using multiparametric flow cytometry. Human microvascular endothelial cells (HMEC-1) and LV epicardial CD105+ non-endothelial cells were used to determine the effect of high glucose on ADAM10-dependent cleavage of ErbB receptors. Results We found that diabetes mellitus (DM) and high levels of hemoglobin A1C are associated with reduced expression of ErbB2 on both endothelial cells and CD105+ non-endothelial cells. To determine if the expression of ErbB2 receptors is regulated by glucose levels, we examined the effect of high glucose in HMEC-1 and LV epicardial CD105+ non-endothelial cells, using a novel flow cytometric approach to simultaneously determine the total level, cell surface expression, and phosphorylation of ErbB2. Incubation of cells in the presence of 25 mM D-glucose resulted in decreased cell surface expression of ErbB2. We also found high expression of a disintegrin and metalloproteinase domain-containing protein 10 (ADAM10) on both endothelial cells and CD105+ non-endothelial cells. Inhibition of ADAM10 prevented the high glucose-dependent decrease in the cell surface expression of ErbB2. Conclusions We suggest that high glucose depresses ErbB receptor signaling in endothelial cells and cardiac progenitor cells via the promotion of ADAM10-dependent cleavage of ErbB2 at the cell surface, thus contributing to vascular dysfunction and adverse remodeling seen in diabetic patients.

Thiamine pyrophosphate diminishes nitric oxide synthesis in endothelial cells

2020 ◽  
pp. 1-9
Author(s):  
Susana Alcázar-Leyva ◽  
Estrella Zapata ◽  
Demetrio Bernal-Alcántara ◽  
Patricia Gorocica ◽  
Noé Alvarado-Vásquez
Keyword(s):  
Nitric Oxide ◽  
Cell Proliferation ◽  
Endothelial Cells ◽  
High Glucose ◽  
Thiamine Pyrophosphate ◽  
Diabetic Patients ◽  
Protective Agent ◽  
Nitric Oxide Synthesis ◽  
Umbilical Cord Vein ◽  
Cells Cultured

Abstract. Although thiamine pyrophosphate (TPP) is considered a protective agent for endothelial cells, it is still unknown if this is associated with nitric oxide (NO) synthesis. Our aim was to evaluate the synthesis of NO in endothelial cells incubated with TPP and high glucose concentrations. Endothelial cells from the umbilical cord vein from newborns (n = 20), were incubated with 5, 15 or 30 mmol/L glucose, in absence or presence of 0.625 mg/ml of TPP. Our results showed a significant increase in cell proliferation (> 40%; P < 0.05), and cell viability (> 90%; P < 0.001) after 48 h in endothelial cells cultured with glucose plus TPP. Likewise, in the presence of glucose and TPP an important rise in the consumption of glucose by the endothelial cells was observed after 24 h (> 7%; P < 0.001) and 48 h (> 10%; P < 0.05). Additionally, the levels of lactate after incubation with glucose and TPP showed only slight variations after 48 h (P < 0.05). However, these changes were clearly different from those observed in the absence of TPP. Interestingly, we found that the changes mentioned were linked with reduced levels of nitrites both at 24 h (< 171 pmol/μg protein; P < 0.001), and 48 h (< 250 pmol/μg protein; P < 0.05), which was associated with a reduced expression of mRNA of eNOS in endothelial cells incubated with TPP and high glucose. In conclusion, the presence of TPP regulates the consumption of glucose and the synthesis of NO, which would explain its protective effect in the endothelium of diabetic patients.

scholarly journals High-glucose-induced metallothionein expression in endothelial cells: an endothelin-mediated mechanism

2001 ◽  
Vol 281 (3) ◽  
pp. C899-C907 ◽  
Author(s):  
Margarita D. Apostolova ◽  
Shali Chen ◽  
Subrata Chakrabarti ◽  
M. George Cherian
Keyword(s):  
Oxidative Stress ◽  
Endothelial Cells ◽  
Mrna Expression ◽  
High Glucose ◽  
Umbilical Vein ◽  
Cell Permeability ◽  
Extracellular Glucose ◽  
Dose Dependent Increase ◽  
Dose Dependent ◽  
Dependent Pathway

Vascular endothelial cells are constantly exposed to oxidative stress and must be protected by physiological responses. In diabetes mellitus, endothelial cell permeability is impaired and may be increased by high extracellular glucose concentrations. It has been postulated that metallothionein (MT) can protect endothelial cells from oxidative stress with its increased expression by cytokines, thrombin, and endothelin (ET)-1. In this study, we demonstrate that high glucose concentration can induce MT expression in endothelial cells through a distinct ET-dependent pathway. Exposure of human umbilical vein endothelial cells (HUVEC) to increasing concentrations of glucose resulted in a rapid dose-dependent increase in MT-2 and ET-1 mRNA expression. MT expression may be further augmented with addition of ET-1. Preincubation of the cells with the specific ETB antagonist BQ-788 blocked MT-2 mRNA expression more effectively than the ETA inhibitor TBC-11251. High glucose also increased immunoreactive MT protein expression and induced translocation of MT into the perinuclear area. Perinuclear localization of MT was related to high-glucose-induced reorganization of F-actin filaments. These results demonstrate that an increase in extracellular glucose in HUVEC can lead to a rapid dose-dependent increase in MT-2 mRNA expression and to perinuclear localization of MT protein with changes to the cytoskeleton. These effects are mediated via the ET receptor-dependent pathway.

scholarly journals Effect of Nicotine and Porphyromonas gingivalis Lipopolysaccharide on Endothelial Cells In Vitro

PLoS ONE ◽  
2014 ◽  
Vol 9 (5) ◽  
pp. e96942 ◽  
Author(s):  
Na An ◽  
Oleh Andrukhov ◽  
Yan Tang ◽  
Frank Falkensammer ◽  
Hans-Peter Bantleon ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Porphyromonas Gingivalis ◽  
Porphyromonas Gingivalis Lipopolysaccharide

scholarly journals Proteasome subunit-α type-6 protein is post-transcriptionally repressed by the microRNA-4490 in diabetic nephropathy

2018 ◽  
Vol 38 (5) ◽  
Author(s):  
Ying Feng ◽  
Ming-yue Jin ◽  
Dong-wei Liu ◽  
Li Wei
Keyword(s):  
Diabetic Nephropathy ◽  
Protein Expression ◽  
Mrna Expression ◽  
High Glucose ◽  
Renal Cortex ◽  
Luciferase Assay ◽  
Diabetic Patients ◽  
Proteasome Subunit ◽  
High Glucose Condition ◽  
Glucose Condition

A common complication of both type I and type II diabetes is nephropathy, characterized by accumulation of extracellular matrix in the glomerular mesangium. This indicates a central role of mesangial cells in the pathophysiology of diabetic nephropathy. Using the proteomic approach, it was earlier elucidated in a rat model that the proteasome subunit-α type-6 protein (PSMA6) is suppressed in the renal cortex in nephropathic kidney. However, the underlying mechanism effecting suppression of PSMA6 protein in the renal cortex is not yet known. Twenty diabetic patients were enrolled and the expression level of PSMA6 in them was detected by immunohistochemistry. The protein and mRNA expression levels of PSMA6 in NRK-52E cells under high glucose condition were determined by Western blot and quantitative real-time PCR, respectively. Dual luciferase assay was used to detect the relationship of PSMA6 and miR-4490. Our results show that PSMA6 protein is down-regulated in patients with diabetic nephropathy compared with healthy control. Using the NRK-52E cell line cultured under high glucose condition as an in vitro model of diabetic nephropathy, we show that loss of PSMA6 protein expression occured independent of changes the in PSMA6 mRNA expression. We next elucidate that PSMA6 mRNA is post-transcriptionally regulated by the microRNA (miRNA)-4490, whose expression is inversely correlated to PSMA6 protein expression. Using reporter assays we show that PSMA6 is a direct target of the miR-4490. Exogenous manipulation of miR-4490 levels modulated expression of PSMA6, indicating that miR-4490 can be tested as a biomarker for nephropathy in diabetic patients.

scholarly journals Potential Effect of Polyphenolic-Rich Fractions of Corn Silk on Protecting Endothelial Cells against High Glucose Damage Using In Vitro and In Vivo Approaches

Molecules ◽  
2021 ◽  
Vol 26 (12) ◽  
pp. 3665
Author(s):  
Nurraihana Hamzah ◽  
Sabreena Safuan ◽  
Wan Rosli Wan Ishak
Keyword(s):  
Endothelial Cells ◽  
Protective Effect ◽  
High Glucose ◽  
Vascular Complications ◽  
Diabetic Rats ◽  
Diabetic Patients ◽  
Dependent Manner ◽  
Corn Silk

Endothelial cell dysfunction is considered to be one of the major causes of vascular complications in diabetes. Polyphenols are known as potent antioxidants that can contribute to the prevention of diabetes. Corn silk has been reported to contain polyphenols and has been used in folk medicine in China for the treatment of diabetes. The present study aims to investigate the potential protective role of the phenolic-rich fraction of corn silk (PRF) against injuries to vascular endothelial cells under high glucose conditions in vitro and in vivo. The protective effect of PRF from high glucose toxicity was investigated using human umbilical vein endothelial cells (HUVECs). The protective effect of PRF was subsequently evaluated by using in vivo methods in streptozotocin (STZ)-induced diabetic rats. Results showed that the PRF significantly reduced the cytotoxicity of glucose by restoring cell viability in a dose-dependent manner. PRF was also able to prevent the histological changes in the aorta of STZ-induced diabetic rats. Results suggested that PRF might have a beneficial effect on diabetic patients and may help to prevent the development and progression of diabetic complications such as diabetic nephropathy and atherosclerosis.

Sign in / Sign up

Export Citation Format

Share Document