Tetramethylpyrazine Attenuates the Endotheliotoxicity and the Mitochondrial Dysfunction by Doxorubicin via 14-3-3γ/Bcl-2

Abstract

Doxorubicin (Dox) with cardiotoxicity and endotheliotoxicity limits its clinical application for cancer. The toxicitic mechanism involves excess ROS generation. 14-3-3s have the protective effects on various injured tissues and cells. Tetramethylpyrazine (TMP) is an alkaloid extracted from the rhizome of Ligusticum wallichii and has multiple bioactivities. We hypothesize that TMP has the protective effects on vascular endothelium by upregulating 14-3-3γ. To test the hypothesis, Dox-induced endotheliotoxicity was used to establish vascular endothelium injury models in mice and human umbilical vein endothelial cells. The effects of TMP were assessed by determining thoracic aortic strips' endothelium-dependent dilation (EDD), as well as LDH, CK, caspase-3, SOD, CAT, GSH-Px activities and MDA level in serum, apoptotic rate, and histopathological changes of vascular tissue (in vivo). Also, cell viability, LDH and caspase-3 activities, ROS generation, levels of NAD+/NADH and GSH/GSSG, MMP, mPTP opening, and apoptotic rate were evaluated (in vitro). The expression of 14-3-3γ and Bcl-2, as well as phosphorylation of Bad (S112), were determined by Western blot. Our results showed that Dox-induced injury to vascular endothelium was decreased by TMP via upregulating 14-3-3γ expression in total protein and Bcl-2 expression in mitochondria, activating Bad (S112) phosphorylation, maintaining EDD, reducing LDH, CK, and caspase-3 activities, thereby causing a reduction in apoptotic rate, and histopathological changes of vascular endothelium (in vivo). Furthermore, TMP increased cell viability and MMP levels, maintained NAD+/NADH, GSH/GSSG balance, decreased LDH and caspase-3 activities, ROS generation, mPTP opening, and apoptotic rate (in vitro). However, the protective effects to vascular endothelium of TMP were significantly canceled by pAD/14-3-3γ-shRNA, an adenovirus that caused knockdown 14-3-3γ expression, or ABT-737, a specific Bcl-2 inhibitor. In conclusion, this study is the first to demonstrate that TMP protects the vascular endothelium against Dox-induced injury via upregulating 14-3-3γ expression, promoting translocation of Bcl-2 to the mitochondria, closing mPTP, maintaining MMP, inhibiting RIRR mechanism, suppressing oxidative stress, improving mitochondrial function, and alleviating Dox-induced endotheliotoxicity.

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.…”

Vinegar/Tetramethylpyrazine Induces Nutritional Preconditioning Protecting the Myocardium Mediated by VDAC1

“…TMP is a promising candidate for managing cardiovascular and cerebrovascular diseases [10][11][12]. Our previous study also articulated that TMP has excellent protective function on a variety of the myocardium and blood vessel injuries [13][14][15][16]. In this study, we characterized the effect of vinegar/TMP pretreatment in alleviating A/R-induced myocardial damage by multiple functional, enzymatic, cellular, or molecular biological indicators (Figures 2-5, Table 2).…”

“…Previously, we have found that TMP has an excellent protection on a variety of the myocardium or blood vessel injuries [13][14][15][16].…”

“…NPC is a convenient and feasible substitute, as it provides similar myocardial protection through dietary intake of natural and nontoxic nutrients [20]. In previous studies, we have demonstrated the use of several nutrients (including TMP, ferulic acid, resveratrol, curcumin, astragaloside IV, and apigenin) for NPC to effectively and safely protect the myocardium and blood vessels against various injuries [14,[20][21][22][23][24]. Therefore, it could be concluded that in this study, the effects of vinegar/TMP pretreatment belong to NPC myocardial protection.…”

Bevacizumab‐Induced Mitochondrial Dysfunction, Endoplasmic Reticulum Stress, and ERK Inactivation Contribute to Cardiotoxicity

“…Previous studies have reported that targeted cancer therapies, such as doxorubicin [ 15 ], anthracycline [ 16 ], and cantharidin [ 17 ], can result in cardiovascular toxicities. The adverse effects of chemotherapy drugs may result from impaired mitochondrial function, increased oxidative stress [ 15 , 18 ], increased mitochondria-proteasome interactions [ 19 ], impaired mitochondrial autophagy [ 20 , 21 ], activation of mitochondrial inflammation signaling pathways such as NF- κ B [ 22 ], mitochondrial energy metabolic dysfunction [ 23 ], and mitochondrial apoptosis [ 24 ]. In addition, ER-mediated abnormalities in intracellular calcium signaling, protein misfolding as a result of ER stress, and ER-dependent cell apoptosis [ 25 – 27 ] can also contribute to cardiomyocyte damage during chemotherapy.…”