Tanshinone IIA Pretreatment Protects H9c2 Cells against Anoxia/Reoxygenation Injury: Involvement of the Translocation of Bcl‐2 to Mitochondria Mediated by 14‐3‐3η

Abstract

Tanshinone IIA is an important component that is isolated from danshen (Salvia miltiorrhiza), which is known to be beneficial for cardiovascular health. In this study, we determined the effects of Tanshinone IIA and its underlying mechanisms of action in an anoxia/reoxygenation (A/R) cell line model. Prior to inducing A/R injury, rat cardiomyocyte-derived cell line H9c2 was stimulated with 8 μM of Tanshinone IIA for 48 hours. When compared with the A/R group, the Tanshinone IIA treatment significantly increased cell viability and decreased lactate dehydrogenase activity. Tanshinone IIA upregulated 14-3-3η expression and facilitated Bcl-2 translocation to the mitochondrial outer membrane, which bound with voltage-dependent anion channel 1. In addition, pretreatment with Tanshinone IIA reduced the generation of reactive oxygen species and cytochrome c release, inactivated caspase-3, prevented mitochondrial permeability transition pore opening, and reduced the percentage of apoptotic cells. Moreover, treatment with Tanshinone IIA reduced the level of malondialdehyde, thereby increasing the activity of superoxide dismutase and glutathione peroxidase. Silencing the expression of 14-3-3η by adenovirus blocked the above-mentioned results. These novel findings showed that pretreatment with Tanshinone IIA alleviated H9c2 cell damage against A/R injury and was associated with upregulation of 14-3-3η, thereby facilitating Bcl-2 translocation to the mitochondrial outer membrane and preventing mitochondrial permeability transition pore opening, decreasing cytochrome c release, preventing caspase-3 activation, and restraining apoptosis.

Doxorubicin Induces Endotheliotoxicity and Mitochondrial Dysfunction via ROS/eNOS/NO Pathway

“…In previous studies, we confirmed that 1 mM Dox for 24 h could induce myocardial cytotoxicity (He et al, 2018). Cell viability and LDH leakage generally serve as indexes of cell injury (Zhang et al, 2018). The MTS assay results showed that the viability of the Dox groups was significantly lower than that of the control group, and LDH activity of the Dox groups was significantly higher relative to that observed in the control group (P < 0.01, Figures 4A, B).…”

Capsaicin Alleviates the Deteriorative Mitochondrial Function by Upregulating 14-3-3η in Anoxic or Anoxic/Reoxygenated Cardiomyocytes

“…14-3-3 is the molecular target of many active ingredients of plants. We have confirmed that 14-3-3 assists PKCε, Bcl-2, and other functional proteins to locate to mitochondria and protect cardiomyocytes and vascular endothelial cells against multiple injuries [7,9,[40][41][42][43]. Further studies are needed to define specific mechanism(s) of action for Cap-activated 14-3-3η in anoxia and A/R injured cardiomyocytes.…”

“…Functionally, together with partner proteins, 14-3-3 regulates phosphorylation and dephosphorylation, kinase activity, and cellular location of proteins that may participate in cell proliferation, differentiation, survival, transformation, and apoptosis [37,38]. Our previous study demonstrated that 14-3-3η is activated in ischemia/hypoxia injury while 14-3-3γ activation is linked to infectious/inflammatory lesions [7][8][9]39]. 14-3-3 is the molecular target of many active ingredients of plants.…”

“…Reactive oxygen species (ROS) participate in several pathophysiologic processes (e.g., cellular damage, aging, and apoptosis) during the above injury [4][5][6]. This injury causes excessive ROS generation, resulting in severe myocardial damage [3][4][5][6][7][8][9]. However, ignoring the close relationship between redox balance and ROS in cellular pathological con-ditions often prevents clinical trials from recognizing the significance of decreasing disease risk and progression.…”

Tetrahydroxystilbene Glucoside Ameliorates Infrasound-Induced Central Nervous System (CNS) Injury by Improving Antioxidant and Anti-Inflammatory Capacity

“…Apoptosis-Related Genes. Bcl-2 [38], BAX [39], and caspase-3 [40,41] have been reported to be associated with oxidative stress and apoptosis. Infrasound treatment reduced relative mRNA level of Bcl-2 and a sharp increase in relative mRNA levels of bax and caspase-3.…”