Nerolidol Attenuates Oxidative Stress, Inflammation, and Apoptosis by Modulating Nrf2/MAPK Signaling Pathways in Doxorubicin-Induced Acute Cardiotoxicity in Rats

Figures

• 

  Figure 1. (A–C) Effect of NERO on myocardial injury markers; columns do not share a common symbol (*, **) and differ significantly from each other (* p < 0.05 vs. normal control, ** p < 0.05 vs. DOX control). (D). Histopathology of the myocardium: Normal control rat’s heart has normal architecture of the myocardium; NERO alone treated rat’s heart also shows normal intact muscle fibers without any pathological changes; DOX-injected rat’s heart shows extensive muscle fiber degradation with inflammatory cells; NERO treated DOX-challenged rats shows a reduced muscle fiber degradation without inflammatory cells. CK- creatine kinase; LDH- lactate dehydrogenase; NOR-normal; NERO-nerolidol; DOX- doxorubicin; ND- rats administered with a single intraperitoneal dose of DOX and orally treated with NERO.

• 

  Figure 2. (A–D) Effect of NERO on lipid peroxidation and oxidative stress. Lipid peroxidation and antioxidant status in the heart. Each column is mean ± SEM for eight rats in each group; columns do not share a common symbol (*, **) and differ significantly with each other (* p < 0.05 vs. normal control, ** p < 0.05 vs. DOX control). (E,F). Western immunoblot analysis and densitometric analysis for SOD1, SOD2, p67-phox, Nrf2, HO-1, and Keap-1. Values are expressed as mean ± SEM (n = 3); columns do not share a common symbol (*, **) and differ significantly with each other (* p < 0.05 vs. normal control, ** p < 0.05 vs. DOX control). MDA- malondialdehyde; SOD 1- superoxide dismutase 1; SOD 2- superoxide dismutase 2; HO-1-heme oxygenase-1; GSH-reduced glutathione; Nrf 2- Nuclear factor erythroid 2-related factor 2; NOR-normal; NERO-nerolidol; DOX- doxorubicin; ND- rats administered with a single intraperitoneal dose of DOX and orally treated with NERO.

• 

  Figure 3. (A–E) Levels/expressions of TNF-α, IL-6, and IL-1β in the myocardium. Each column is mean ± SEM for eight rats in each group for ELISA and 3 rats in each group for immunoblotting. Columns do not share a common symbol (*, **) and differ significantly with each other (* p < 0.05 vs. normal control, ** p < 0.05 vs. DOX control). Representative images of Western immunoblot analysis and densitometric analysis for TNF-α, IL-6, and IL-1β in the myocardium. TNF-α- tumor necrosis factor α; IL-6- interleukin 6; IL-1β- interleukin 1β; NOR-normal; NERO-nerolidol; DOX- doxorubicin; ND- rats administered with a single intraperitoneal dose of DOX and orally treated with NERO.

• 

  Figure 4. (A,B) Western immunoblotting and densitometric analysis for iNOS, COX-2, NF-κB-P65, p-NF-κBP65, IκBα and p-IκBα, t-IKKα, p-IKKα, t-JNK, p-JNK, P38, and p-P38. The values are expressed as the mean ± SEM (n = 3); columns do not share a common symbol (*, **) and differ significantly with each other (* p < 0.05 vs. normal control, ** p < 0.05 vs. DOX control).iNOS-inducible nitric oxide synthase; COX-2- cyclooxygenase-2; p-NF-κB-p65- phospho nuclear factor kappa-B-p65; t-JNK- t-Jun-amino-terminal kinase; p-JNK- p-Jun-amino-terminal kinase; p-IKKα- IκB kinase-α; P38-tumor protein; p-P38-phosphorilated p38; NOR-normal; NERO-nerolidol; DOX- doxorubicin; ND- rats administered with a single intraperitoneal dose of DOX and orally treated with NERO.

• 

  Figure 5. (A,B). Representative images of Western immunoblot analysis and densitometry for γ-H2AX, P53, Bax, Bcl2, Bcl-xL, active caspase-3, active caspase-9, APAF-1, cleaved PARP, and cytochrome-C. The values are expressed as the mean ± SEM (n = 3); columns do not share a common symbol (*, **) and differ significantly with each other (* p < 0.05 vs. normal control, ** p < 0.05 vs. DOX control). H2AX- H2A histone family member X; Bcl-2- B-cell lymphoma 2; Bax- Bcl2-associated-X- protein; Bcl-xL- B-cell lymphoma extra-large; P53- tumor protein; APAF-1- apoptotic protease activating factor 1; Cyt-C- cytochrome-C; CL-PARP-cleaved poly (ADP-ribose) polymerase; Casp-3-caspase 3; Cl-Casp-3- cleaved caspase-3; Casp-9-caspase 9; Cl-Casp-9- cleaved caspase-9; NOR-normal; NERO-nerolidol; DOX- doxorubicin; ND- rats administered with a single intraperitoneal dose of DOX and orally treated with NERO.