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

Binger Lu,Yicun Chen,Yanmei Zhang,Bin Wang,Fuchun Zheng,Ganggang Shi,Shuping Zhong,Fenfei Gao

doi:10.18632/oncotarget.9186

Abstract

Endothelial cells are highly sensitive to hypoxia and contribute to myocardial ischemia/reperfusion injury. We have reported that N-n-butyl haloperidol iodide (F2) can attenuate hypoxia/reoxygenation (H/R) injury in cardiac microvascular endothelial cells (CMECs). However, the molecular mechanisms remain unclear. Neonatal rat CMECs were isolated and subjected to H/R. Pretreatment of F2 leads to a reduction in H/R injury, as evidenced by increased cell viability, decreased lactate dehydrogenase (LDH) leakage and apoptosis, together with enhanced AMP-activated protein kinase (AMPK) and liver kinase B1 (LKB1) phosphorylation in H/R ECs. Blockade of AMPK with compound C reversed F2-induced inhibition of H/R injury, as evidenced by decreased cell viability, increased LDH release and apoptosis. Moreover, compound C also blocked the ability of F2 to reduce H/R-induced reactive oxygen species (ROS) generation. Supplementation with the ROS scavenger N-acetyl-L-cysteine (NAC) reduced ROS levels, increased cell survival rate, and decreased both LDH release and apoptosis after H/R. In conclusion, our data indicate that F2 may mitigate H/R injury by stimulating LKB1/AMPK signaling pathway and subsequent suppression of ROS production in CMECs.

Highlights

Myocardial ischemia/reperfusion (I/R) injury is a universal cardiovascular disease with major cause of morbidity, mortality all around the world and great cost to the society [1]
We have reported that N-n-butyl haloperidol iodide (F2) can attenuate hypoxia/reoxygenation (H/R) injury in cardiac microvascular endothelial cells (CMECs)
CMECs are sensitive to hypoxic injury, which influences the prognosis, development, and pathogenesis of cardiovascular disease [29]

Summary

Introduction

Myocardial ischemia/reperfusion (I/R) injury is a universal cardiovascular disease with major cause of morbidity, mortality all around the world and great cost to the society [1]. Cardiac microvascular endothelial cells (CMECs) play a pivotal role in the development, contractile performance, and rhythmicity of heart [2]. It is well known that the endothelium, especially the microvascular endothelium, is highly susceptible to hypoxia and plays a crucial role during I/R injury [3]. During hypoxia/reoxygenation (H/R), endothelial cells exhibit a proinflammatory phenotype, including the induction of vasoconstrictive agents, leukocyte adhesion molecules and procoagulant factors [4]. Rats subjected to I/R injury manifest increased apoptosis of CMECs [5]. Therapeutic strategies focusing on the maintenance of endothelial cell function have been shown to alleviate myocardial injury [4, 6]

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