N-n-Butyl Haloperidol Iodide Ameliorates Oxidative Stress in Mitochondria Induced by Hypoxia/Reoxygenation through the Mitochondrial c-Jun N-Terminal Kinase/Sab/Src/Reactive Oxygen Species Pathway in H9c2 Cells

Abstract

Both c-Jun N-terminal kinase (JNK) and reactive oxygen species (ROS) play important roles in myocardial ischemia/reperfusion (I/R) injury. Our previous studies suggest that N-n-butyl haloperidol iodide (F2) exerts cardioprotection by reducing ROS production and JNK activation caused by I/R. In this study, we hypothesized that there is a JNK/Sab/Src/ROS pathway in the mitochondria in H9c2 cells following hypoxia/reoxygenation (H/R) that induces oxidative stress in the mitochondria and that F2 exerts mitochondrial protective effects during H/R injury by modulating this pathway. The results showed that H/R induced higher-level ROS in the cytoplasm on the one hand and JNK activation and translocation to the mitochondria by colocalization with Sab on the other. Moreover, H/R resulted in mitochondrial Src dephosphorylation, and subsequently, oxidative stress evidenced by the increase in ROS generation and oxidized cardiolipin in the mitochondrial membranes and by the decrease in mitochondrial superoxide dismutase activity and membrane potential. Furthermore, treatment with a JNK inhibitor or Sab small interfering RNA inhibited the mitochondrial translocation of p-JNK, decreased colocalization of p-JNK and Sab on the mitochondria, and reduced Src dephosphorylation and mitochondrial oxidative stress during H/R. In addition, Src dephosphorylation by inhibitor PP2 increased mitochondrial ROS production. F2, like inhibitors of the JNK/Sab/Src/ROS pathway, downregulated the H/R-induced mitochondrial translocation of p-JNK and the colocalization of p-JNK and Sab on the mitochondria, increased Src phosphorylation, and alleviated the above-mentioned mitochondrial oxidative stress. In conclusion, F2 could ameliorate H/R-associated oxidative stress in mitochondria in H9c2 cells through the mitochondrial JNK/Sab/Src/ROS pathway.

Src-family Protein Tyrosine Kinases: A promising target for treating Cardiovascular Diseases

“…In other words, Src tyrosine kinase was inactivated in MIRI [66]. phosphorylated Src expression in the mitochondria decreased with an increased mitochondrial ROS level after H/R treatment through JNK/Sab/Src/ROS pathway [67]. And, the activation of Src concentrates on various components of the reperfusion injury salvage kinase pathway that has been proved as a potential target for protecting the heart from ischemic/reperfusion during the period of ischemic preconditioning [68].…”

Wenxin Granule Ameliorates Hypoxia/Reoxygenation-Induced Oxidative Stress in Mitochondria via the PKC-δ/NOX2/ROS Pathway in H9c2 Cells

“…Mitochondria are abundantly expressed in the myocardium as an energy supporter for myocardial contraction [51,52]. Mitochondrial homeostasis is considered a vital target in cardiac I/R therapy, due to their insufficient supply of energy, excessive generation of ROS, and releasing cytochrome C and other proapoptotic factors [53,54]. We investigated whether PKC-δ/NOX2derived ROS contributed to the stimulation of mitochondrial dysfunction in response to H/R treatments in H9c2 cells.…”

Dexmedetomidine Protects Human Cardiomyocytes Against Ischemia-Reperfusion Injury Through α2-Adrenergic Receptor/AMPK-Dependent Autophagy

“…Another concern is how the in vitro H/R process imitates the in vivo I/R process. According to current publications, the H/R models used for in vitro study have varied (Riquelme et al, 2016;Wang et al, 2018;Chu et al, 2019;Fernández et al, 2020;Ma et al, 2020 Jan). They can be divided into two categories.…”

“…They can be divided into two categories. One is physical hypoxia, which places the cells under a hypoxic condition (Wang et al, 2018;Chu et al, 2019;Ma et al, 2020 Jan). The other is chemical hypoxia, in which oxygen scavengers deplete oxygen Gao et al, 2020).…”