The protective effects of Egr-1 antisense oligodeoxyribonucleotide on cardiac microvascular endothelial injury induced by hypoxia–reoxygenationThis paper is one of a selection of papers published in this special issue entitled “Second International Symposium on Recent Advances in Basic, Clinical, and Social Medicine” and has undergone the Journal's usual peer review process.

Abstract

Early growth response 1 (Egr-1) over-expression has been demonstrated in myocardial ischemia-reperfusion injury, which is closely associated with endothelial dysfunction. In the present study we investigated the expression of Egr-1 on cultured cardiac microvascular endothelial cells (CMECs) to help define the mechanism of myocardial ischemia-reperfusion injury. A model of cultured CMECs exposed to hypoxia-reoxygenation was developed in which synthesized Egr-1 sense and antisense oligodeoxyribonucleotide were transfected into the cells. The expression of Egr-1 was examined by Western blot analysis. Lactate dehydrogenase, malondialdehyde, superoxide dismutase, tumor necrosis factor alpha, and intercellular adhesion molecule 1 were measured after hypoxia-reoxygenation to assess cell function and injury. Cell morphology, cell viability, and neutrophil adhesion to the CMECs were measured to assess the degree of injury and inflammation. Only cells transfected with Egr-1 antisense oligodeoxyribonucleotide showed a significant reduction in Egr-1 protein expression following hypoxia-reoxygenation. Consistent with the down-regulation of Egr-1 expression, other forms of cell injury were significantly reduced in this group of cells, as evidenced by less alteration in cell morphology, a decrease in expression of tumor necrosis factor alpha and intercellular adhesion molecule 1, improved cell survival, and reduced neutrophil adhesion.

N-n-butyl Haloperidol Iodide Protects against Hypoxia/Reoxygenation Injury in Cardiac Microvascular Endothelial Cells by Regulating the ROS/MAPK/Egr-1 Pathway

“…Recent studies by our laboratory suggest that oxidative stress (induced by excessive ROS) and Egr-1 expression are key factors responsible for H/R damage in CMECs. Furthermore, ROS and Egr-1, might crosstalk to mediate H/R injury of CMECs (Zhou et al, 2010a,b; Zhang et al, 2015). Demonstration that an activated ROS/Egr-1 pathway is behind the mechanism of H/R injury in CMECs would be another important step for prevention and therapy of myocardial I/R injury.…”

“…The study was approved by the Institutional Animal Care and Use Committee of Shantou University Medical College. Rat CMECs were isolated as described (Zhou et al, 2010a) and cultured in DMEM supplemented with 10% FBS, ECGS (15 mg/L) and heparin sodium (6.25 U/ml) at 37°C under 5% CO 2 . CMECs were identified by antibody against CD31, which is constitutively expressed on the surface of CMECs.…”

“…CMECs were identified by antibody against CD31, which is constitutively expressed on the surface of CMECs. Hypoxia was induced as before with some modifications (Zhou et al, 2010a). Briefly, CMECs were cultured in hypoxia solution in an air-tight chamber saturated with pure N 2 at 37°C for 1, 2, 3, or 4 h; the culture conditions were then returned to normal for 1 h of reoxygenation.…”

“…Previous studies on CMECs, as well as H9c2 cells, revealed that oxidative stress and Egr-1 expression contribute to H/R injury (Zhou et al, 2010b). Moreover, damage resulting from oxidative stress decreases in the presence of Egr-1 antisense oligonucleotide, indicating that the ROS/Egr-1 pathway might exist in CMECs and be responsible for H/R injury in CMECs and even heart tissue (Zhou et al, 2010a). However, a ROS/Egr-1 pathway in CMECs has yet to be identified.…”

N-n-butyl haloperidol iodide ameliorates hypoxia/reoxygenation injury through modulating the LKB1/AMPK/ROS pathway in cardiac microvascular endothelial cells

“…CMECs were isolated from the left ventricles of hearts from neonatal rats, as previously described [40], with slight modifications. Briefly, the left ventricles were finely minced and digested with 0.1% trypsin for twice, for 5 min at 37°C each time, in a shaking water bath.…”