Effect of N-n-butyl haloperidol iodide on ROS/JNK/Egr-1 signaling in H9c2 cells after hypoxia/reoxygenation

Abstract

Reactive oxygen species (ROS)-induced oxidative stress in cells is an important pathophysiological process during myocardial ischemia/reperfusion (I/R) injury, and the transcription factor Egr-1 is a master switch for various damage pathways during reperfusion injury. An in vitro model of myocardial I/R injury and H9c2 cardiomyoblast cells hypoxia/reoxygenation (H/R) was used to assess whether there is abnormal intracellular ROS/JNK/Egr-1 signaling. We also assessed whether N-n-butyl haloperidol (F2), which exerts protective effects during myocardial I/R injury, can modulate this pathway. H/R induced ROS generation, JNK activation, and increased the expression of Egr-1 protein in H9c2 cells. The ROS scavengers edaravone (EDA) and N-acetyl-L-cysteine (NAC) reduced ROS level, downregulated JNK activation, and Egr-1 expression in H9c2 cells after H/R. The JNK inhibitor SP600125 inhibited Egr-1 overexpression in H9c2 cells caused by H/R. F2 could downregulate H/R-induced ROS level, JNK activation, and Egr-1 expression in H9c2 cells in a dose-dependent manner. The ROS donor hypoxanthine-xanthine oxidase (XO/HX) and the JNK activator ANISO antagonized the effects of F2. Therefore, H/R activates ROS/Egr-1 signaling pathway in H9c2 cells, and JNK activation plays an important role in this pathway. F2 regulates H/R-induced ROS/JNK/Egr-1 signaling, which might be an important mechanism by which it antagonizes myocardial I/R injury.

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

“…Our former investigations have shown that the cardioprotection of F 2 may be linked to its capacity block calcium channels [20–22] and inhibition of the overexpression of early growth response gene-1, a major switch for multiple pathways of reperfusion injury [23–25]. Consistent with the attenuation of myocardial injury, we have also demonstrated that F 2 inhibits H/R-induced ROS generation in cardiomyocytes [26]. However, related studies of F 2 on cultured CMECs in vitro are limited.…”

“…The Increased ROS levels in the vasculature results in endothelial damage [37], which acts as the major etiological factor underlying I/R injury [38]. We showed in our former study that F 2 reduces the production of ROS after H/R in cardiomyocytes, which is accompanied by improved myocardial function [26]. Consistent with these findings, our present data demonstrated that F 2 dose-dependently suppresses the generation of ROS in H/R-challenged CMECs.…”

Age-associated up-regulation of EGR1 promotes granulosa cell apoptosis during follicle atresia in mice through the NF-κB pathway

“…31 A recent report revealed a convincing correlation between ROS and EGR1 expression and showed that intracellular ROS levels up-regulate EGR1 expression, and those researchers considered that EGR1 might act as the "damage hub" to mediate oxidative stress and the related ROS-induced damage during ischemia and reperfusion. 32 It has been established that the increase in ROS-induced damage during aging plays a vital role in ovarian aging. 33 The present study showed that EGR1 plays a role in promoting apoptosis in GCs under H 2 O 2 stimulation.…”

A microfluidic-based lid device for conventional cell culture dishes to automatically control oxygen level

“…However, conventional hypoxic cell culture devices, including a modular chamber or a gas incubator, must be opened again after hypoxic conditions have been established for experimental manipulations, such as confluency check, medium replacement and cytokine solution infusion [2]. Hypoxic cells eventually undergo reoxygenation when the culture dish is exposed to the normoxia atmosphere, resulting in reactive oxygen species generation and cell damage [3,4]. A glove box allows the manipulation of the culture dish without exposing it to the atmosphere, but it is very expensive and large; hence, it is often not available in small-scale laboratories [2].…”