Allopurinol protects human glomerular endothelial cells from high glucose-induced reactive oxygen species generation, p53 overexpression and endothelial dysfunction

Metformin is a first-line drug for the management of type 2 diabetes. Recent studies suggested cardioprotective effects of metformin against ischemia/reperfusion injury. However, it remains elusive whether metformin provides direct protection against hypoxia/reoxygenation (H/R) injury in cardiomyocytes under normal or hyperglycemic conditions. This study in H9C2 rat cardiomyoblasts was designed to determine cell viability under H/R and high-glucose (HG, 33 mM) conditions and the effects of cotreatment with various concentrations of metformin (0, 1, 5, and 10 mM). We further elucidated molecular mechanisms underlying metformin-induced cytoprotection, especially the possible involvement of AMP-activated protein kinase (AMPK) and Jun NH(2)-terminal kinase (JNK). Results indicated that 5 mM metformin improved cell viability, mitochondrial integrity, and respiratory chain activity under HG and/or H/R (P<0.05). The beneficial effects were associated with reduced levels of reactive oxygen species generation and proinflammatory cytokines (TNF-α, IL-1α, and IL-6) (P<0.05). Metformin enhanced phosphorylation level of AMPK and suppressed HG + H/R induced JNK activation. Inhibitor of AMPK (compound C) or activator of JNK (anisomycin) abolished the cytoprotective effects of metformin. In conclusion, our study demonstrated for the first time that metformin possessed direct cytoprotective effects against HG and H/R injury in cardiac cells via signaling mechanisms involving activation of AMPK and concomitant inhibition of JNK.

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Ethyl Rosmarinate Protects High Glucose-Induced Injury in Human Endothelial Cells

Molecules ◽

10.3390/molecules23123372 ◽

2018 ◽

Vol 23 (12) ◽

pp. 3372 ◽

Cited By ~ 3

Author(s):

Yan-Hui Shen ◽

Li-Ying Wang ◽

Bao-Bao Zhang ◽

Qi-Ming Hu ◽

Pu Wang ◽

...

Keyword(s):

Reactive Oxygen Species ◽

Endothelial Cells ◽

Protective Effect ◽

High Glucose ◽

Reactive Oxygen ◽

Annexin V ◽

Ros Generation ◽

Dependent Manner ◽

Oxygen Species ◽

Jnk Pathway

Ethyl rosmarinate (RAE) is one of the active constituents from Clinopodium chinense (Benth.) O. Kuntze, which is used for diabetic treatment in Chinese folk medicine. In this study, we investigated the protective effect of RAE on high glucose-induced injury in endothelial cells and explored its underlying mechanisms. Our results showed that both RAE and rosmarinic acid (RA) increased cell viability, decreased the production of reactive oxygen species (ROS), and attenuated high glucose-induced endothelial cells apoptosis in a dose-dependent manner, as evidenced by Hochest staining, Annexin V–FITC/PI double staining, and caspase-3 activity. RAE and RA both elevated Bcl-2 expression and reduced Bax expression, according to Western blot. We also found that LY294002 (phosphatidylinositol 3-kinase, or PI3K inhibitor) weakened the protective effect of RAE. In addition, PDTC (nuclear factor-κB, or NF-κB inhibitor) and SP600125 (c-Jun N-terminal kinase, or JNK inhibitor) could inhibit the apoptosis in endothelial cells caused by high glucose. Further, we demonstrated that RAE activated Akt, and the molecular docking analysis predicted that RAE showed more affinity with Akt than RA. Moreover, we found that RAE inhibited the activation of NF-κB and JNK. These results suggested that RAE protected endothelial cells from high glucose-induced apoptosis by alleviating reactive oxygen species (ROS) generation, and regulating the PI3K/Akt/Bcl-2 pathway, the NF-κB pathway, and the JNK pathway. In general, RAE showed greater potency than RA equivalent.

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Aspirin-triggered lipoxin A4 blocks reactive oxygen species generation in endothelial cells: A novel antioxidative mechanism

Thrombosis and Haemostasis ◽

10.1160/th06-06-0315 ◽

2007 ◽

Vol 97 (01) ◽

pp. 88-98 ◽

Cited By ~ 78

Author(s):

Christina Barja-Fidalgo ◽

Vany Nascimento-Silva ◽

Maria Arruda ◽

Iolanda Fierro

Keyword(s):

Reactive Oxygen Species ◽

Endothelial Cells ◽

Cardiovascular Diseases ◽

Reactive Oxygen Species Generation ◽

Reactive Oxygen ◽

Protective Role ◽

Ros Generation ◽

Oxygen Species ◽

Lipoxin A4 ◽

Pre Treatment

SummaryLipoxins and their aspirin-triggered carbon-15 epimers have emerged as mediators of key events in endogenous anti-inflammation and resolution. However, the implication of these novel lipid mediators on cardiovascular diseases such as hypertension, atherosclerosis, and heart failure has not been investigated. One of the major features shared by these pathological conditions is the increased production of reactive oxygen species (ROS) generated by vascular NAD(P)H oxidase activation. In this study, we have examined whether an aspirin-triggered lipoxin A4 analog (ATL-1) modulates ROS generation in endothelial cells (EC). Pre-treatment of EC with ATL-1 (1–100 nM) completely blocked ROS production triggered by different agents, as assessed by dihydrorhodamine 123 and hydroethidine. Furthermore, ATL-1 inhibited the phosphorylation and translocation of the cytosplamic NAD(P)H oxidase subunit p47phox to the cell membrane as well as NAD(P)H oxidase activity. Western blot and immunofluorescence microscopy analyses showed that ATL-1 (100 nM) impaired the redox-sensitive activation of the transcriptional factor NF-κB, a critical step in several events associated to vascular pathologies. These results demonstrate that ATL-1 suppresses NAD(P)H oxidase-mediated ROS generation in EC, strongly indicating that lipoxins may play a protective role against the development and progression of cardiovascular diseases.

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Probucol promotes high glucose-induced proliferation and inhibits apoptosis by reducing reactive oxygen species generation in Müller cells

International Ophthalmology ◽

10.1007/s10792-019-01130-8 ◽

2019 ◽

Vol 39 (12) ◽

pp. 2833-2842 ◽

Cited By ~ 3

Author(s):

Xuxia Zhou ◽

ShiBei Ai ◽

ZhongPing Chen ◽

ChenXiang Li

Keyword(s):

Reactive Oxygen Species ◽

High Glucose ◽

Reactive Oxygen Species Generation ◽

Reactive Oxygen ◽

Müller Cells ◽

Oxygen Species ◽

Muller Cells

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Adiponectin Suppression of High-Glucose-Induced Reactive Oxygen Species in Vascular Endothelial Cells: Evidence for Involvement of a cAMP Signaling Pathway

Diabetes ◽

10.2337/db05-1174 ◽

2006 ◽

Vol 55 (6) ◽

pp. 1840-1846 ◽

Cited By ~ 181

Author(s):

R. Ouedraogo ◽

X. Wu ◽

S.-Q. Xu ◽

L. Fuchsel ◽

H. Motoshima ◽

...

Keyword(s):

Reactive Oxygen Species ◽

Endothelial Cells ◽

Signaling Pathway ◽

High Glucose ◽

Vascular Endothelial Cells ◽

Reactive Oxygen ◽

Camp Signaling ◽

Vascular Endothelial ◽

Oxygen Species ◽

Camp Signaling Pathway

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High-Glucose-Induced Reactive Oxygen Species (ROS) Generation Is Reduced by Microtubular Agents in Microvascular Endothelial Cells

Free Radical Biology and Medicine ◽

10.1016/j.freeradbiomed.2014.10.231 ◽

2014 ◽

Vol 76 ◽

pp. S110

Author(s):

Domokos Gero ◽

Csaba Szabo

Keyword(s):

Reactive Oxygen Species ◽

Endothelial Cells ◽

High Glucose ◽

Reactive Oxygen ◽

Microvascular Endothelial Cells ◽

Ros Generation ◽

Oxygen Species ◽

Microvascular Endothelial

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Activation of NF-κB induced by H2O2 and TNF-α and its effects on ICAM-1 expression in endothelial cells

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.2000.279.2.l302 ◽

2000 ◽

Vol 279 (2) ◽

pp. L302-L311 ◽

Cited By ~ 87

Author(s):

Andrea L. True ◽

Arshad Rahman ◽

Asrar B. Malik

Keyword(s):

Gene Expression ◽

Reactive Oxygen Species ◽

Endothelial Cells ◽

Nuclear Dna ◽

Reactive Oxygen Species Generation ◽

Reactive Oxygen ◽

Intercellular Adhesion Molecule ◽

Oxygen Species ◽

Tnf Α

Reactive oxygen species have been proposed to signal the activation of the transcription factor nuclear factor (NF)-κB in response to tumor necrosis factor (TNF)-α challenge. In the present study, we investigated the effects of H2O2 and TNF-α in mediating activation of NF-κB and transcription of the intercellular adhesion molecule (ICAM)-1 gene. Northern blot analysis showed that TNF-α exposure of human dermal microvascular endothelial cells (HMEC-1) induced marked increases in ICAM-1 mRNA and cell surface protein expression. In contrast, H2O2 added at subcytolytic concentrations failed to activate ICAM-1 expression. Challenge with H2O2 also failed to induce NF-κB-driven reporter gene expression in the transduced HMEC-1 cells, whereas TNF-α increased the NF-κB-driven gene expression ∼10-fold. Gel supershift assay revealed the presence of p65 (Rel A), p50, and c-Rel in both H2O2- and TNF-α-induced NF-κB complexes bound to the ICAM-1 promoter, with the binding of the p65 subunit being the most prominent. In vivo phosphorylation studies, however, showed that TNF-α exposure induced marked phosphorylation of NF-κB p65 in HMEC-1 cells, whereas H2O2 had no effect. These results suggest that reactive oxygen species generation in endothelial cells mediates the binding of NF-κB to nuclear DNA, whereas TNF-α generates additional signals that induce phosphorylation of the bound NF-κB p65 and confer transcriptional competency to NF-κB.

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