CYP2J2-derived epoxyeicosatrienoic acids protect against doxorubicin-induced cardiotoxicity by reducing oxidative stress and apoptosis via activation of the AMPK pathway

Figures

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  Fig. 1. Overexpression of CYP2J2 protected against cardiac dysfunction in DOX-treated mice. (A, B) Body weight and HW/TL ratio of mice in each group. (C–G) Representative echocardiographic and hemodynamic analyses in mice. (H, I) Relative cardiac mRNA levels of ANP and BNP in mice. Data are presented as the mean ± SEM (n = 7 per group). *P < 0.05 vs. GFP; #P < 0.05 vs. DOX + GFP.

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  Fig. 2. Overexpression of CYP2J2 alleviated cardiac atrophy and activated AMPK pathway in DOX-treated mice. (A, B) HE and Masson's trichrome staining of heart sections from mice. (C–E) Serum levels of cTnT, CK-MB, LDH in mice were determined. (F) Representative immunoblots and quantitative analysis of p-AMPKα, AMPKα, PGC-1α, and UCP2 in the hearts from different groups. Data are presented as the mean ± SEM (n = 7 per group). *P < 0.05 vs. GFP; #P < 0.05 vs. DOX + GFP.

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  Fig. 3. Overexpression of CYP2J2 decreased cardiac oxidative stress, mitochondrial dysfunction, and apoptosis in DOX-treated mice. (A) Representative images of DHE staining of cardiac ROS production. (B) MDA content, GSH level, and SOD enzyme activity in the heart. (C) Representative immunoblots and quantitative analysis of SOD1, SOD2, NOX2 and NOX4 levels in the heart from different groups. (D) Representative immunoblots and quantitative analysis of Bcl-2, Bax, C-caspase-3, and caspase-3 levels in the heart from different groups. Data are presented as the mean ± SEM (n = 7 per group). *P < 0.05 vs. GFP; #P < 0.05 vs. DOX + GFP.

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  Fig. 4. 14,15-EET suppressed oxidative stress, mitochondrial dysfunction, and apoptosis in DOX-exposed H9C2 cells. (A) DHE staining of H9C2 cells. (B) Representative immunoblots and quantitative analysis of SOD1, SOD2, NOX2, and NOX4 levels in H9c2 cells. (C) H9C2 cells apoptosis were detected via flow cytometry by use of Annexin V/PI staining. (D) Representative immunoblots and quantitative analysis of Bcl-2, Bax, C-caspase-3, and Caspase-3 expression in H9C2 cells. (E) Representative immunoblots and quantitative analysis of p-AMPKα, AMPKα, PGC-1α, and UCP2 levels in H9C2 cells. Data are presented as mean ± SEM (n = 3 per group). *P < 0.05 vs. DMSO; #P < 0.05 vs. DOX; &P < 0.05 vs. DOX+14,15-EET.

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  Fig. 5. 14,15-EET decreases oxidative stress, mitochondrial dysfunction and apoptosis in DOX-incubated H9C2 cells through the AMPK pathway. (A) DHE staining of H9C2 cells. (B) Representative immunoblots and quantitative analysis of SOD1, SOD2, NOX2, NOX4 levels in H9C2 cells. (C) H9C2 cell apoptosis was detected via flow cytometry by use of Annexin V/PI staining. (D) Representative immunoblots and quantitative analysis of Bcl-2, Bax, C-caspase-3, and Caspase-3 levels in H9C2 cells. (E) Representative immunoblots and quantitative analysis of PGC-1α, UCP2 expression levels in H9C2 cells. Data are presented as mean ± SEM (n = 3 per group). *P < 0.05 vs. si-con; #P < 0.05 vs. DOX + si-con; &P < 0.05 vs. DOX+14,15-EET + si-con.