Matrine attenuates oxidative stress and cardiomyocyte apoptosis in doxorubicin-induced cardiotoxicity via maintaining AMPK α/UCP2 pathway

Can Hu; Xin Zhang; Wenying Wei; Ning Zhang; Haiming Wu; Zhenguo Ma; Lingli Li; Wei Deng; Qizhu Tang

doi:10.1016/j.apsb.2019.03.003

Matrine attenuates oxidative stress and cardiomyocyte apoptosis in doxorubicin-induced cardiotoxicity via maintaining AMPK α/UCP2 pathway

Acta Pharm Sin B. 2019 Jul;9(4):690-701. doi: 10.1016/j.apsb.2019.03.003. Epub 2019 Mar 16.

Authors

Can Hu^{1

2

3}, Xin Zhang^{1

2

3}, Wenying Wei^{1

2

3}, Ning Zhang^{1

2

3}, Haiming Wu^{1

2

3}, Zhenguo Ma^{1

2

3}, Lingli Li^{1

2

3}, Wei Deng^{1

2

3}, Qizhu Tang^{1

2

3}

Affiliations

¹ Department of Cardiology, Renmin Hospital of Wuhan University, Wuhan 430060, China.
² Cardiovascular Research Institute of Wuhan University, Wuhan 430060, China.
³ Hubei Key Laboratory of Cardiology, Wuhan 430060, China.

Abstract

Oxidative stress and cardiomyocyte apoptosis are involved in the pathogenesis of doxorubicin (DOX)-induced cardiotoxicity. Matrine is well-known for its powerful anti-oxidant and anti-apoptotic capacities. Our present study aimed to investigate the effect of matrine on DOX-induced cardiotoxicity and try to unearth the underlying mechanisms. Mice were exposed with DOX to generate DOX-induced cardiotoxicity or normal saline as control. H9C2 cells were used to verify the effect of matrine in vitro. DOX injection triggered increased generation of reactive oxygen species (ROS) and excessive cardiomyocyte apoptosis, which were significantly mitigated by matrine. Mechanistically, we found that matrine ameliorated DOX-induced uncoupling protein 2 (UCP2) downregulation, and UCP2 inhibition by genipin could blunt the protective effect of matrine on DOX-induced oxidative stress and cardiomyocyte apoptosis. Besides, 5'-AMP-activated protein kinase α2 (Ampkα2) deficiency inhibited matrine-mediated UCP2 preservation and abolished the beneficial effect of matrine in mice. Besides, we observed that matrine incubation alleviated DOX-induced H9C2 cells apoptosis and oxidative stress level via activating AMPKα/UCP2, which were blunted by either AMPKα or UCP2 inhibition with genetic or pharmacological methods. Matrine attenuated oxidative stress and cardiomyocyte apoptosis in DOX-induced cardiotoxicity via maintaining AMPKα/UCP2 pathway, and it might be a promising therapeutic agent for the treatment of DOX-induced cardiotoxicity.

Keywords: 4-HNE, 4-hydroxynonenal; ACC, acetyl-CoA carboxylase; AMPKα; AMPKα, 5′-AMP-activated protein kinase α; ANOVA, analysis of variance; Apoptosis; BAX, BCL-2-associated X protein; BCA, bicinchoninic acid; BCL-2, B-cell lymphoma 2; C-caspase 3, cleaved-caspase3; CCK-8, cell counting kit 8; CK-MB, creatine kinase isoenzymes; DCFH-DA, 2′,7′-dichlorodihydrofluorescein diacetate; DHE, dihydroethidium; DMEM, Dulbecco׳s modified Eagle׳s medium; DOX, doxorubicin; FBS, fetal bovine serum; FS, fractional shortening; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; HW, heart weight; LDH, lactate dehydrogenase; MDA, malondialdehyde; Matrine; Oxidative stress; PPARs, peroxisomal proliferators-activated receptors; ROS, reactive oxygen species; SOD2, superoxide dismutase 2; T-caspase3, total-caspase3; TL, tibia length; TUNEL, TdT-mediated dUTP nick end-labelling; Top2, topoisomerase-II; UCP2; UCP2, uncoupling protein 2; cTnT, cardiac isoform of Tropnin T.