Protective Effects of Oxymatrine on Vascular Endothelial Cells from High-Glucose-Induced Cytotoxicity by Inhibiting the Expression of A2B Receptor

Background/Aims: Diabetes mellitus (DM) has become an increasingly epidemic metabolic disease. Vascular endothelial cells play a key role in developing the cardiovascular complications of DM. The A2B receptor is expressed in vascular endothelial cells, and may help regulate the function of endothelial cells. The aim of this study was to investigate the protective effects of oxymatrine (OMT) on human umbilical vein endothelial cells (HUVECs) from high glucose-induced cytotoxicity. Methods: Homology modeling and molecular docking analysis were used to detect the binding sites between the adenosine A2B receptor and OMT. HUVECs were cultured with control (5.5 mM) or elevated glucose (22.2 mM) in the presence or absence of 3 µM OMT or A2B siRNA for 3 days. The MTS cell viability assay was used to measure the toxicity of high glucose on HUVECs and the protective effect of OMT or A2B siRNA. The expression of the adenosine A2B receptor and CCL5 in HUVECs was detected with real-time quantitative PCR (qPCR) and Western blotting methods in each group. Levels of IL-1β and TNF-α were measured using an enzyme-linked immunosorbent assay (ELISA) kit, and the concentration of NO was detected with the nitrate reductase method. Monocyte chemotactic activity in each group was detected using Transwell chambers. Furthermore, the phosphorylation of p38 and ERK1/2 in each group was observed through the Western blotting method. Results: Homology modeling and molecular docking analysis showed that OMT contains well-fitted binding sites to the A2B receptor. After chronic culture at high glucose, the rate of cell viability was significantly lower than that of the control group. After co-treatment with OMT or A2B siRNA, cell viability was significantly increased compared with the high-glucose group. The results from real-time quantitative RT-PCR (qRT-PCR) and Western blotting indicated that high glucose could increase the expression of A2B receptors in HUVECs, an effect that was inhibited by OMT. In addition, the results revealed that the expression of CCL5, IL-1β and TNF-α was increased in the high-glucose group, and that the NO produced by HUVECs decreased due to hyperglycemia; however, co-culture with OMT or A2B siRNA abolished these effects. Meanwhile, the chemotaxis activity of monocytes to HUVECs cultured in high-glucose medium was enhanced 2.59-fold compared to the control cells. However, the inflammatory reactions in HUVECs were completely relieved by co-treatment with OMT or A2B siRNA. Moreover, the phosphorylation of p38 and ERK1/2 in HUVECs in the high-glucose group was significantly higher than that of the control group; these effects were reversed after co-treatment with OMT or A2B siRNA. Conclusion: OMT may protect the HUVECs from high glucose-induced cytotoxicity through inhibitting the expression of A2B receptor and inflammatory factors as well as decreasing the phosphorylation of p38 and ERK1/2.

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High glucose enhances CD39 expression in vascular endothelial cells

Asian Biomedicine ◽

10.5372/1905-7415.0802.291 ◽

2014 ◽

Vol 8 (2) ◽

pp. 283-287

Author(s):

Sudawadee Kongkhum ◽

Mudtika Fungkrajai ◽

Sompoch Prajan ◽

Narisa Kengtrong Bordeerat ◽

Kanyanath Piumngam ◽

...

Keyword(s):

Endothelial Cells ◽

Blood Glucose ◽

Cell Viability ◽

High Glucose ◽

Inflammatory Process ◽

Vascular Endothelial Cells ◽

Vascular Endothelial ◽

High Blood Glucose ◽

Glucose Levels ◽

Blood Glucose Levels

Abstract Background: Diabetes mellitus (DM) patients lose their ability to control normal blood glucose levels, resulting in high blood glucose levels (hyperglycemia). Hyperglycemia causes DM complications. This involves responses of vascular endothelial cells (VECs) to hyperglycemia, affecting inflammatory process and platelet activity. Ecto-enzyme CD39 is expressed on VECs, catalyzing the hydrolysis of ATP and ADP to AMP and, consequently, regulating inflammatory process and platelet activation. Objective: We studied whether high glucose concentration has an effect on CD39 expression on VECs. Methods: Cultured human umbilical vein endothelial cells (HUVEC) were used as a model of study. HUVEC were cultured in different glucose conditions (4, 9, 24, and 34 mM) for 24 hours. Cell viability was assessed using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT)-based assay and expression of CD39 was examined by using SDS-PAGE and western blot techniques. Results: HUVEC were cultured in normal (4 and 9 mM) or high (24 and 34 mM) glucose concentrations for short term (24 hours). The results showed that high glucose (24 and 34 mM) reduced cell viability to 89.5 ± 11.3 and 86.3 ± 13.5 (mean ± SD), compared with control (4 mM), respectively. High glucose also induced increases in CD39 expression in HUVEC. Conclusion: High glucose decreases cell viability and increases CD39 expression in HUVEC, suggesting involvement of CD39 in cell responses to high glucose.

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Curcumin suppresses oxidative stress via regulation of ROS/NF-κB signaling pathway to protect retinal vascular endothelial cell in diabetic retinopathy

10.21203/rs.3.rs-27239/v1 ◽

2020 ◽

Author(s):

Jiang Huang ◽

Quanyong Yi ◽

Yao Chen ◽

Yi Li ◽

Guoxu Xu ◽

...

Keyword(s):

Oxidative Stress ◽

Flow Cytometry ◽

Endothelial Cells ◽

Diabetic Retinopathy ◽

High Glucose ◽

Glucose Concentration ◽

Vascular Endothelial Cells ◽

Control Group ◽

Vascular Endothelial ◽

High Glucose Concentration

Abstract Background: Diabetic retinopathy (DR) is one of the most serious complications of diabetes mellitus and a leading blindness disease in the world. The retinal vascular endothelial cells can be damaged by oxidative stress even in the early stage of diabetic retinopathy. NF-κB is a key transcription factor in cell apoptosis and oxidative stress. Curcumin can relieve oxidative stress induced by high glucose. This study aimed to investigate the effect of curcumin on the rat retinal vascular endothelial cells (RRVECs) in DR and to deduce the possible molecular mechanism. Methods: The cultured RRVECs were identified by both of vWF and CD31 expression.The RRVECs were divided into four groups: the normal control group, the osmolarity control group, the high glucose group and the curcumin treatment group (High glucose +Curcumin). We observed the different morphological changes in the groups by transmission electron microscopy. Oxidative stress was detected by flow cytometry. The Activation of ROS/NF-κB signal pathway was detected by electrophoretic Mobility Shift Assay (EMSA), immunohistochemistry and western-blot; the apoptosis of RRVECs was tested by flow cytometry. Results: We found that curcumin reduced the reactive oxygen species (ROS) and relieved the apoptosis in RRVECs exposed to the high glucose by flow cytometry. We detected that the increased activity of NF-κB and phosphorylated NF-κB in RRVECs induced by high glucose concentration was significantly suppressed by curcumin. Furthermore, based on the assay quantifying the level of apoptosis-related proteins, including bcl-2 and bax, we illustrated that curcumin could decrease the apoptosis of RRVECs induced by high glucose concentration. Conclusion: We concluded that ROS/NF-κB signaling pathway played an important role in the progress of DR and curcumin could suppress the oxidative stress via regulation of NF-κB signal to protect the RRVECs in DR.

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TPTEP1 suppresses high glucose-induced dysfunction in retinal vascular endothelial cells by interacting with STAT3 and targeting VEGFA

Acta Diabetologica ◽

10.1007/s00592-020-01663-w ◽

2021 ◽

Author(s):

Xiaoping Sun ◽

Yuebing Lu ◽

Tao Lei

Keyword(s):

Endothelial Cells ◽

High Glucose ◽

Vascular Endothelial Cells ◽

Vascular Endothelial

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The Use of Fiber-Reinforced Scaffolds Cocultured with Schwann Cells and Vascular Endothelial Cells to Repair Rabbit Sciatic Nerve Defect with Vascularization

BioMed Research International ◽

10.1155/2013/362918 ◽

2013 ◽

Vol 2013 ◽

pp. 1-7 ◽

Cited By ~ 4

Author(s):

Hongyang Gao ◽

Yang You ◽

Guoping Zhang ◽

Feng Zhao ◽

Ziyi Sha ◽

...

Keyword(s):

Endothelial Cells ◽

Sciatic Nerve ◽

Nerve Fiber ◽

Schwann Cells ◽

Vascular Endothelial Cells ◽

Neurological Function ◽

Control Group ◽

Vascular Endothelial ◽

Fiber Reinforced ◽

3D Scaffolds

To explore the feasibility of biodegradable fiber-reinforced 3D scaffolds with satisfactory mechanical properties for the repair of long-distance sciatic nerve defect in rabbits and effects of vascularized graft in early stage on the recovery of neurological function, Schwann cells and vascular endothelial cells were cocultured in the fiber-reinforced 3D scaffolds. Experiment group which used prevascularized nerve complex for the repair of sciatic nerve defect and control group which only cultured with Schwann cells were set. The animals in both groups underwent electromyography to show the status of the neurological function recovery at 4, 8, and 16 weeks after the surgery. Sciatic nerve regeneration and myelination were observed under the light microscope and electron microscope. Myelin sheath thickness, axonal diameter, and number of myelinated nerve fiber were quantitatively analyzed using image analysis system. The recovery of foot ulcer, the velocity of nerve conduction, the number of regenerating nerve fiber, and the recovery of ultrastructure were increased in the experimental group than those in the control group. Prevascularized tissue engineered fiber-reinforced 3D scaffolds for the repair of sciatic nerve defects in rabbits can effectively promote the recovery of neurological function.

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