scholarly journals Exosomal circHIPK3 Released from Hypoxia-Pretreated Cardiomyocytes Regulates Oxidative Damage in Cardiac Microvascular Endothelial Cells via the miR-29a/IGF-1 Pathway

2019 ◽  
Vol 2019 ◽  
pp. 1-28 ◽  
Author(s):  
Yan Wang ◽  
Ranzun Zhao ◽  
Weiwei Liu ◽  
Zhenglong Wang ◽  
Jidong Rong ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Endothelial Cells ◽  
Ectopic Expression ◽  
Bioactive Molecules ◽  
Microvascular Endothelial Cells ◽  
Circular Rnas ◽  
Protective Effects ◽  
Microvascular Endothelial ◽  
Cardiac Microvascular Endothelial Cells

Background/Aims. Circular RNAs (circRNAs) are a class of endogenous noncoding RNAs that regulate gene expression in eukaryotes. Recently, exosomes from cardiomyocytes (CMs) have been found to facilitate cell proliferation and survival by transporting various bioactive molecules, including circRNA. However, the functions of exosomal circRNAs are not clear. The present research is aimed at determining whether circHIPK3 released from hypoxia-pretreated CMs is transferred into cardiac microvascular endothelial cells (CMVECs) by exosomes and becomes functionally active in the CMVECs under oxidative stress conditions. Methods. Quantitative polymerase chain reactions were conducted to detect the expression pattern of circHIPK3 in CMVECs under oxidative stress. Annexin V-FITC/propidium iodide (PI) staining assays, TUNEL assays, ROS assays, and Western blot analysis were conducted to detect the role of exosomal circHIPK3 in CMVEC function in vitro. Luciferase activity assays and RNA immunoprecipitation studies were conducted in vitro to reveal the mechanism of circHIPK3-mediated CMVEC function. Results. circHIPK3 expression was significantly upregulated in hypoxic exosomes (HPC-exos) compared with normoxic exosomes (Nor-exos). Moreover, HPC-exos induced stronger antioxidant effects than Nor-exos. The silencing or overexpression of circHIPK3 changed CMVEC survival under oxidative conditions in vitro. Furthermore, circHIPK3 silencing in HPC-exos abrogated the protective effects of HPC-exos in CMVECs, as shown by increased levels of apoptosis, ROS, MDA, and proapoptotic proteins. circHIPK3 acted as an endogenous miR-29a sponge to sequester and inhibit miR-29a activity, which led to increased IGF-1 expression. The ectopic expression of miR-29a mimicked the effect of circHIPK3 silencing in CMVECs in vitro. Conclusions. circHIPK3 in HPC-exos plays a role in CMVECs under oxidative conditions through miR-29a-mediated IGF-1 expression, leading to a decrease in oxidative stress-induced CMVECs dysfunction. These data suggest that the exosomal circRNA in CMs is a potential target to control CMVECs dysfunction under oxidative conditions.

Isolation and characterization of human and rat cardiac microvascular endothelial cells

1993 ◽  
Vol 264 (2) ◽  
pp. H639-H652 ◽  
Author(s):  
M. Nishida ◽  
W. W. Carley ◽  
M. E. Gerritsen ◽  
O. Ellingsen ◽  
R. A. Kelly ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Endothelial Cell ◽  
Microvascular Endothelial Cells ◽  
Microvascular Endothelium ◽  
Adult Rat ◽  
Isolation And Characterization ◽  
Ventricular Tissue ◽  
Microvascular Endothelial ◽  
Cardiac Microvascular Endothelial Cells

Although reciprocal intercellular signaling may occur between endocardial or microvascular endothelium and cardiac myocytes, suitable in vitro models have not been well characterized. In this report, we describe the isolation and primary culture of cardiac microvascular endothelial cells (CMEC) from both adult rat and human ventricular tissue. Differential uptake of fluorescently labeled acetylated low-density lipoprotein (Ac-LDL) indicated that primary isolates of rat CMEC were quite homogeneous, unlike primary isolates of human ventricular tissue, which required cell sorting based on Ac-LDL uptake to create endothelial cell-enriched primary cultures. The endothelial phenotype of both primary isolates and postsort subcultured CMEC and their microvascular origin were determined by characteristic histochemical staining for a number of endothelial cell-specific markers, by the absence of cells with fibroblast or pericyte-specific cell surface antigens, and by rapid tube formation on purified basement membrane preparations. Importantly, [3H]-thymidine uptake was increased 2.3-fold in subconfluent rat microvascular endothelial cells 3 days after coculture with adult rat ventricular myocytes because of release of an endothelial cell mitogen(s) into the extracellular matrix, resulting in a 68% increase in cell number compared with CMEC in monoculture. Thus biologically relevant cell-to-cell interactions can be modeled with this in vitro system.

scholarly journals Lipoic acid antagonizes paraquat-induced vascular endothelial dysfunction by suppressing mitochondrial reactive oxidative stress

2019 ◽  
Vol 8 (6) ◽  
pp. 918-927 ◽  
Author(s):  
Li Pang ◽  
Ping Deng ◽  
Yi-dan Liang ◽  
Jing-yu Qian ◽  
Li-Chuan Wu ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Nitric Oxide ◽  
Endothelial Cells ◽  
Endothelial Dysfunction ◽  
Lipoic Acid ◽  
Microvascular Endothelial Cells ◽  
Ros Generation ◽  
Protective Effects ◽  
Migration Ability ◽  
Microvascular Endothelial

Abstract Paraquat (PQ) is a widely used herbicide in the agricultural field. The lack of an effective antidote is the significant cause of high mortality in PQ poisoning. Here, we investigate the antagonistic effects of alpha lipoic acid (α-LA), a naturally existing antioxidant, on PQ toxicity in human microvascular endothelial cells (HMEC-1). All the doses of 250, 500 and 1000 μM α-LA significantly inhibited 1000 μM PQ-induced cytotoxicity in HMEC-1 cells. α-LA pretreatment remarkably diminished the damage to cell migration ability, recovered the declined levels of the vasodilator factor nitric oxide (NO), elevated the expression level of endothelial nitric oxide synthases (eNOS), and inhibited the upregulated expression of vasoconstrictor factor endothelin-1 (ET-1). Moreover, α-LA pretreatment inhibited reactive oxygen species (ROS) generation, suppressed the damage to the mitochondrial membrane potential (ΔΨm) and mitigated the inhibition of adenosine triphosphate (ATP) production in HMEC-1 cells. These results suggested that α-LA could alleviate PQ-induced endothelial dysfunction by suppressing oxidative stress. In summary, our present study provides novel insight into the protective effects and pharmacological potential of α-LA against PQ toxicity in microvascular endothelial cells.

scholarly journals Protective Effects of Scutellarin on Human Cardiac Microvascular Endothelial Cells against Hypoxia-Reoxygenation Injury and Its Possible Target-Related Proteins

2015 ◽  
Vol 2015 ◽  
pp. 1-13 ◽  
Author(s):  
Meina Shi ◽  
Yingting Liu ◽  
Lixing Feng ◽  
Yingbo Cui ◽  
Yajuan Chen ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Expression Profiles ◽  
Interaction Network ◽  
Treated Group ◽  
Microvascular Endothelial Cells ◽  
Protective Effects ◽  
Microvascular Endothelial ◽  
Related Proteins ◽  
Cardiac Microvascular Endothelial Cells ◽  
Hypoxia Reoxygenation

Scutellarin (SCU) is one of the main components of traditional Chinese medicine plantErigeron breviscapus (Vant.)Hand.-Mazz. In this paper, we studied the protective effects of SCU on human cardiac microvascular endothelial cells (HCMECs) against hypoxia-reoxygenation (HR) injury and its possible target-related proteins. Results of MTT assay showed that pretreatment of SCU at doses of 1, 5, and 10 μM for 2 h could significantly inhibit the decrease in cell viability of HCMECs induced by HR injury. Subcellular fractions of cells treated with vehicle control, 1 μM SCU, HR injury, or 1 μM SCU + HR injury were separated by ultracentrifugation. The protein expression profiles of cytoplasm and membrane/nuclei fractions were checked using protein two-dimensional electrophoresis (2-DE). Proteins differentially expressed between control and SCU-treated group, control and HR group, or HR and SCU + HR group were identified using mass spectrometry (MS/MS). Possible interaction network of these target-related proteins was predicted using bioinformatic analysis. The influence of SCU on the expression levels of these proteins was confirmed using Western blotting assay. The results indicated that proteins such as p27BBP protein (EIF6), heat shock 60 kDa protein 1 (HSPD1), and chaperonin containing TCP1 subunit 6A isoform (CCT6A) might play important roles in the effects of SCU.

Reaktion der Endothelzellen auf kurzzeitig physiologisch erhöhte hydrostatische Drücke und oxidativen Stress in vitro

2018 ◽  
Vol 236 (09) ◽  
pp. 1122-1128
Author(s):  
Carolina Mann ◽  
Solon Thanos ◽  
Katrin Brockhaus ◽  
Franz H. Grus ◽  
Norbert Pfeiffer ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Endothelial Cells ◽  
Endotheliale Dysfunktion ◽  
Microvascular Endothelial Cells ◽  
Brain Microvascular Endothelial Cells ◽  
Microvascular Endothelial

Zusammenfassung Hintergrund Im Rahmen der Pathogenese des Glaukoms wird die endotheliale Dysfunktion zunehmend diskutiert. Peripapilläre Blutungen sind diagnostisch wegweisend. Die Korrelation von Glaukomerkrankungen mit vaskulärem Dysregulationssyndrom ist eindeutig. Ziel dieser Studie ist es, die genaue Reaktion der Endothelzellen auf erhöhten hydrostatischen und oxidativen Stress zu untersuchen. Material und Methoden In vitro wurden primär dissoziierte BMECs (brain microvascular endothelial cells) für 3 Tage normalem und leicht erhöhtem hydrostatischem Druck von 60 und 120 mmHg in einer Druckkammer ausgesetzt. Zusätzlich wurden sowohl druckbelastete als auch nicht druckbelastete Zellen oxidativem Stress in Form von geringen Konzentrationen H2O2 ausgesetzt. Ein Live/Dead Assay wurde durchgeführt, um die Zellviabilität zu messen. Morphologisch wurden die Zellen mit immunhistochemischer Aktinfärbung beurteilt. Ergebnisse Interessanterweise zeigten die Endothelzellen sowohl unter 60 mmHg als auch unter 120 mmHg kein vermehrtes Absterben im Vergleich zu den Zellen ohne Belastung. Auch morphologisch zeigten sich keine großen Unterschiede. Gegenüber oxidativem Stress wurden alle Zellen schon bei kleinen Mengen geschädigt. Keinen Unterschied konnte man zwischen oxidativem Stress ohne vorherige Druckbelastung und oxidativem Stress mit vorheriger Druckbelastung von 120 mmHg für 3 Tage feststellen. Schlussfolgerung Wir konnten keinen direkten Effekt in Form von vermehrtem Zelluntergang der Endothelzellen auf erhöhten hydrostatischen Druck feststellen. Allerdings zeigt die Reaktion auf die geringen Konzentrationen von oxidativem Stress, dass die Zellen im Rahmen der Pathogenese des Glaukoms doch in Mitleidenschaft gezogen werden. Der oxidative Stress scheint hier eine besondere Rolle zu spielen.

scholarly journals Aspirin Enhances the Protection of Hsp90 from Heat-Stressed Injury in Cardiac Microvascular Endothelial Cells Through PI3K-Akt and PKM2 Pathways

Cells ◽  
2020 ◽  
Vol 9 (1) ◽  
pp. 243 ◽  
Author(s):  
Xiaohui Zhang ◽  
Bixia Chen ◽  
Jiaxin Wu ◽  
Junzhou Sha ◽  
Bo Yang ◽  
...  
Keyword(s):  
Heart Failure ◽  
Endothelial Cells ◽  
Heat Stress ◽  
Resistance Mechanism ◽  
Resistance Mechanisms ◽  
Microvascular Endothelial Cells ◽  
Microvascular Endothelial Cell ◽  
Microvascular Endothelial ◽  
Cardiac Microvascular Endothelial Cells

Heat stress (HS) often causes sudden death of humans and animals due to heart failure, mainly resulting from the contraction of cardiac microvasculature followed by myocardial ischemia. Cardiac microvascular endothelial cells (CMVECs) play an important role in maintaining vasodilatation. Aspirin (ASA) is well known for its protective abilities of febrile animals. However, there is little knowledge about molecular resistance mechanisms of CMVECs and which role ASA may play in this context. Therefore, we used a heat stress model of rat cardiac microvascular endothelial cell cultures in vitro and investigated the cell injuries and molecular resistance mechanism of CMVECs caused by heat stress, and the effect of aspirin (ASA) on it. HS induced severe pathological damage of CMVECs and cellular oxidative stress and dysfunction of NO release. Hsp90 was proven to be indispensable for resisting HS-injury of CMVECs through PI3K-Akt and PKM2 signaling pathways. Meanwhile, PKM2 functioned in reducing Akt phosphorylation. ASA treatment of CMVECs induced a significant expression of Hsp90, which promoted both Akt and PKM2 signals, which are beneficial for relieving HS damage and maintaining the function of CMVECs. Akt activation also promoted HSF-1 that regulates the expression of Hsp70, which is known to assist Hsp90′s molecular chaperone function and when released to the extracellular liquid to protect myocardial cells from HS damage. To the best of our knowledge, this is the first study to show that HS damages CMVECs and the protection mechanism of Hsp90 on it, and that ASA provides a new potential strategy for regulating cardiac microcirculation preventing HS-induced heart failure.

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