Backgrounds.
Doxorubicin (DOX) is an effective therapeutic
drug for malignant
tumors; however, its clinical applications were limited by its side effects, especially the
cardiotoxicity caused by ROS-mediated p53 and MAPK signal pathways' activation-induced cell apoptosis.
Sanyang Xuedai mixture (
SYKT) has been reported as an
antioxidant agent and attenuated DOX-induced
cardiotoxicity by targeting ROS-mediated apoptosis, but the mechanisms are still not fully delineated. Objective. This study aimed at investigating whether
SYKT alleviated DOX-induced
cardiotoxicity by inhibiting ROS-mediated apoptosis and elucidating the role of ROS-mediated p53 and MAPK signal pathways' activation in this process. Materials and Methods. Identification, separation, and culture of mouse primary cardiomyocytes. Cells were treated with DOX (1 μM),
SYKT (30 mg/mL), or
SYKT coupled with DOX. The p53 inhibitor
Pifithrin-α (PFT-α),
p38/MAPK inhibitor SB203583 (SB), and JNK inhibitor
SP600125 (SP) were used as positive control. Western blot was employed to detected p53 and p38 as well as JNK expressions and the activation and translocation of Bax and
cytochrome C. Flow cytometer (FCM) was used to detect the mitochondrial membrane potential and cell apoptosis. Results. After separation and culture, 95% of cells showed positive cTnI expression, which indicated that mouse primary cardiomyocytes were successfully identified in our research. DOX activated p53 and MAPK signal pathways in a time-dependent manner, which were inactivated by being cotreated with
SYKT, PFT-α, or SB, respectively. DOX significantly decreased Bax and increased
cytochrome c expressions in the cytoplasm, whereas Bax was upregulated and
cytochrome c was downregulated in the mitochondria, which were reversed by
SYKT treatment. Besides, DOX reduced mitochondria membrane potential (
MMP) in cardiomyocytes compared to the control group;
SYKT recovered its
MMP and attenuated DOX-induced cardiomyocyte injury. Of note, DOX increased the expression levels of cleaved
caspase-3 as well as
poly ADP-ribose polymerase (PARP) and promoted cell apoptosis, which were also reversed by
SYKT treatment. Discussion and Conclusions. Our results indicated that
SYKT alleviated DOX-induced
cardiotoxicity by inhibiting p53 and MAPK signal pathways' activation-mediated apoptosis, and it might serve as a potential therapeutic agent for DOX-induced
cardiotoxicity.