Click
here to close Hello! We notice that
you are using Internet Explorer, which is not supported by Echinobase
and may cause the site to display incorrectly. We suggest using a
current version of Chrome,
FireFox,
or Safari.
Mar Drugs
2018 Dec 20;171:. doi: 10.3390/md17010002.
Show Gene links
Show Anatomy links
Spinochrome D Attenuates Doxorubicin-Induced Cardiomyocyte Death via Improving Glutathione Metabolism and Attenuating Oxidative Stress.
Yoon CS
,
Kim HK
,
Mishchenko NP
,
Vasileva EA
,
Fedoreyev SA
,
Stonik VA
,
Han J
.
???displayArticle.abstract???
Doxorubicin, an anthracycline from Streptomyces peucetius, exhibits antitumor activity against various cancers. However, doxorubicin is cardiotoxic at cumulative doses, causing increases in intracellular reactive oxygen species in the heart. Spinochrome D (SpD) has a structure of 2,3,5,6,8-pentahydroxy-1,4-naphthoquinone and is a structural analogue of well-known sea urchin pigment echinochrome A. We previously reported that echinochrome A is cardioprotective against doxorubicin toxicity. In the present study, we assessed the cardioprotective effects of SpD against doxorubicin and determined the underlying mechanism. ¹H-NMR-based metabolomics and mass spectrometry-based proteomics were utilized to characterize the metabolites and proteins induced by SpD in a human cardiomyocyte cell line (AC16) and human breast cancer cell line (MCF-7). Multivariate analyses identified 12 discriminating metabolites (variable importance in projection > 1.0) and 1814 proteins from SpD-treated AC16 cells. Proteomics and metabolomics analyses showed that glutathione metabolism was significantly influenced by SpD treatment in AC16 cells. SpD treatment increased ATP production and the oxygen consumption rate in D-galactose-treated AC16 cells. SpD protected AC16 cells from doxorubicin cytotoxicity, but it did not affect the anticancer properties. With SpD treatment, the mitochondrial membrane potential and mitochondrial calcium localization were significantly different between cardiomyocytes and cancer cell lines. Our findings suggest that SpD could be cardioprotective against the cytotoxicity of doxorubicin.
Figure 2. Mass spectrometry-based proteomics of SpD-treated AC16 cells. Liquid Chromatography–Mass Spectrometry-MS (LC-MS/MS) spectrometry-based proteomics detected proteins from SpD (10 μM, 24 h) (A) and SpD/doxorubicin (B) treated AC16 cells; (C) Search Tool for the Retrieval of Interacting Genes/Proteins (STRING) analysis showed that altered metabolic proteins clustered around “mitochondria” which are represented as red colored nodes. All filled nodes represent the 3D structures of proteins are known; and, (D) The top 10 influenced metabolic pathways are shown from the STRING analysis (Kyoto Encyclopedia of Genes and Genomes (KEGG) database).
Figure 3. Normalization of 1H-NMR aquired metabolite concentrations. The concentrations of metabolites were normalized by log-transformation followed by Pareto scaling (mean-centered and divided by the square root of the standard deviation of each variable). Changes of metabolites are represented as ratios of control metabolites.
Figure 4. Volcano plots for SpD-induced metabolic changes compared with controls (n = 3). Metabolites are considered significant if log2(fold change) > 1.2. The p-value threshold was 0.05. The significantly changed metabolites included acetate, glutamine, myo-inositol, glutathione, taurine, O-phosphocholine, and sn-Glycero-3-phosphocholine (GPC).
Figure 5. 1H-NMR metabolomics for SpD-treated AC16 cells. (A) Principal component analysis (PCA) indicated that metabolites from the SpD-treated (10 μM, 24 h) group were significantly different from those in the control group; (B) Heat-map analysis of metabolites with variable importance in projection (VIP) score > 1.0. The logarithmic fold changes are shown below. GPC, sn-glycero-3-phosphocholine; (C) The loading plots from orthogonal partial least-squares discriminant analysis (OPLS-DA) for SpD metabolites compared with the control group.
Figure 6. Network analysis of metabolites altered by SpD (10 μM, 24 h) treatment of AC16 cells. Networks of metabolites according to their Pearson’s correlation coefficients were drawn using Cytoscape program. The networks with significantly increased GPC and decreased acetate are marked as red lines. (A) control and (B) SpD-treated cell metabolites; and, (C) Pathway impact analysis shows the most affected metabolic pathways affected by SpD. Varying colors from yellow to red represent metabolites’ significance in the data.
Figure 8. SpD caused increased ATP production and oxygen consumption rate (OCR) in AC16 cells. (A) SpD (10 μM) increased ATP production in AC16 cells. D-galactose (10 mM) was added to reduce cytosolic glycolytic ATP production. By changing energy metabolism in cardiomyocytes by replacing glucose with galactose, high concentrations of galactose could prevent ATP production except that of mitochondria by oxidative phosphorylation (OXPHOS); (B) SpD (10 μM) increased OCR. Antimycin A (Ant, 1 μM, a Complex III inhibitor) was used as a cell-based negative control and glucose oxidase (GOx, 1 mg/mL) was used as a cell-free positive control. (C) SpD increased ATP levels under H2O2 induced oxidative stress. (D) SpD increased ATP production in the presence of doxorubicin (0.1 μM). * p < 0.05 compared with untreated controls, # p < 0.05 compared with the D-galactose-treated group.
Figure 9. SpD attenuated doxorubicin-induced mitochondrial membrane potential and mitochondrial calcium changes in AC16 cells. (A) Mitochondrial membrane potential (Δψm) in AC16 cells was indicated by TMRE fluorescence. The cells were treated with doxorubicin (0.1–1.0 μM) with/without SpD (10 μM); (B) The intensity of tetramethylrhodamine (TMRE) staining was measured using fluorometry at 550Ex/590Em nm. Doxorubicin decreased mitochondrial membrane potential in a dose-dependent manner. Co-treatment with SpD attenuated the membrane potential loss; (C) Mitochondrial calcium was localized using rhod-2, a selective indicator for mitochondrial Ca2+; and, (D) The intensity of rhod-2 was measured at 552Ex/581Em nm. Doxorubicin induced diffusion of mitochondrial Ca2+ to the cytosolic space but SpD co-treatment attenuated the Ca2+ diffusion. * p < 0.05 compared with the doxorubicin-treated group.
Adams,
Cardiomyocyte apoptosis induced by Galphaq signaling is mediated by permeability transition pore formation and activation of the mitochondrial death pathway.
2000, Pubmed
Adams,
Cardiomyocyte apoptosis induced by Galphaq signaling is mediated by permeability transition pore formation and activation of the mitochondrial death pathway.
2000,
Pubmed
Agafonova,
Evaluation of effects of histochrome and mexidol on structural and functional characteristics of the brain in senescence-accelerated OXYS rats by magnetic resonance imaging.
2011,
Pubmed
Aguer,
Galactose enhances oxidative metabolism and reveals mitochondrial dysfunction in human primary muscle cells.
2011,
Pubmed
Al-Ghamdi,
Time and dose dependent study of doxorubicin induced DU-145 cytotoxicity.
2008,
Pubmed
Anderson,
Distribution of spinochrome pigments in echinoids.
1969,
Pubmed
,
Echinobase
Arcamone,
Adriamycin, 14-hydroxydaunomycin, a new antitumor antibiotic from S. peucetius var. caesius.
1969,
Pubmed
Bachur,
Nuclear catalyzed antibiotic free radical formation.
1982,
Pubmed
Chhunchha,
Curcumin abates hypoxia-induced oxidative stress based-ER stress-mediated cell death in mouse hippocampal cells (HT22) by controlling Prdx6 and NF-κB regulation.
2013,
Pubmed
Cortés-Funes,
Role of anthracyclines in the era of targeted therapy.
2007,
Pubmed
Diogo,
Cardiac mitochondrial dysfunction during hyperglycemia--the role of oxidative stress and p66Shc signaling.
2013,
Pubmed
Fakhruddin,
Diabetes-Induced Reactive Oxygen Species: Mechanism of Their Generation and Role in Renal Injury.
2017,
Pubmed
Feng,
CaMKII is a nodal signal for multiple programmed cell death pathways in heart.
2017,
Pubmed
Franco,
Anticancer and cardio-protective effects of liposomal doxorubicin in the treatment of breast cancer.
2018,
Pubmed
Hosios,
Acetate metabolism in cancer cells.
2014,
Pubmed
Jeong,
Echinochrome A protects mitochondrial function in cardiomyocytes against cardiotoxic drugs.
2014,
Pubmed
,
Echinobase
Kase,
Remodeling of oxidative energy metabolism by galactose improves glucose handling and metabolic switching in human skeletal muscle cells.
2013,
Pubmed
Knowles,
Production and utilization of acetate in mammals.
1974,
Pubmed
Kotze,
A novel untargeted metabolomics correlation-based network analysis incorporating human metabolic reconstructions.
2013,
Pubmed
Kraskiewicz,
Partial XBP1 knockdown does not affect viability of oligodendrocyte precursor cells exposed to new models of hypoxia and ischemia in vitro.
2011,
Pubmed
Kuznetsov,
Changes in mitochondrial redox state, membrane potential and calcium precede mitochondrial dysfunction in doxorubicin-induced cell death.
2011,
Pubmed
Lane,
Mitochondrial respiration and redox coupling in articular chondrocytes.
2015,
Pubmed
Lebedev,
Antioxidant properties, autooxidation, and mutagenic activity of echinochrome a compared with its etherified derivative.
2001,
Pubmed
Lebedev,
Echinochrome, a naturally occurring iron chelator and free radical scavenger in artificial and natural membrane systems.
2005,
Pubmed
Lefrak,
A clinicopathologic analysis of adriamycin cardiotoxicity.
1973,
Pubmed
Magnelli,
Phorbol esters attenuate the expression of p53 in cells treated with doxorubicin and protect TS-P53/K562 from apoptosis.
1995,
Pubmed
McCurrach,
bax-deficiency promotes drug resistance and oncogenic transformation by attenuating p53-dependent apoptosis.
1997,
Pubmed
Mohamed,
Protective effect of nebivolol on doxorubicin-induced cardiotoxicity in rats.
2018,
Pubmed
Nagaoka,
Poor prognosis of colorectal cancer in patients over 80 years old is associated with down-regulation of tumor suppressor genes.
2003,
Pubmed
Pang,
Hyperglycemia inhibits capacitative calcium entry and hypertrophy in neonatal cardiomyocytes.
2002,
Pubmed
Pilco-Ferreto,
Influence of doxorubicin on apoptosis and oxidative stress in breast cancer cell lines.
2016,
Pubmed
Pozharitskaya,
Antiallergic effects of pigments isolated from green sea urchin (Strongylocentrotus droebachiensis) shells.
2013,
Pubmed
,
Echinobase
Rajaram,
Potential benefits and harms of NADPH oxidase type 4 in the kidneys and cardiovascular system.
2019,
Pubmed
Singh,
Hyperglycemia-induced cardiac contractile dysfunction in the diabetic heart.
2018,
Pubmed
Sinha,
Adriamycin-stimulated hydroxyl radical formation in human breast tumor cells.
1987,
Pubmed
Siveski-Iliskovic,
Probucol protects against adriamycin cardiomyopathy without interfering with its antitumor effect.
1995,
Pubmed
Studneva,
Protective Effects of a Novel Agonist of Galanin Receptors Against Doxorubicin-Induced Cardiotoxicity in Rats.
2019,
Pubmed
Takemura,
Doxorubicin-induced cardiomyopathy from the cardiotoxic mechanisms to management.
2007,
Pubmed
Thorn,
Doxorubicin pathways: pharmacodynamics and adverse effects.
2011,
Pubmed
Volpe,
Cellular death, reactive oxygen species (ROS) and diabetic complications.
2018,
Pubmed
Wang,
Doxorubicin induces apoptosis in normal and tumor cells via distinctly different mechanisms. intermediacy of H(2)O(2)- and p53-dependent pathways.
2004,
Pubmed
Worley,
Multivariate Analysis in Metabolomics.
2013,
Pubmed
Wu,
Hypoxia-induced autophagy contributes to the invasion of salivary adenoid cystic carcinoma through the HIF-1α/BNIP3 signaling pathway.
2015,
Pubmed
