Title:iTRAQ-based Proteomic Analysis Unveils NCAM1 as a Novel Regulator in Doxorubicin-induced Cardiotoxicity and DT-010-exerted Cardioprotection
Volume: 20
Issue: 9
Author(s): Sijie Wang, Caipeng Xie, Huihui Hu, Pei Yu, Haijing Zhong*, Yuqiang Wang*Luchen Shan*
Affiliation:
- State Key Laboratory of Bioactive Molecules and Druggability Assessment, Jinan University, International Cooperative
Laboratory of Traditional Chinese Medicine Modernization and Innovative Drug Development of Chinese Ministry
of Education, Institute of New Drug Research, Jinan University College of Pharmacy, Guangzhou, China
- State Key Laboratory of Bioactive Molecules and Druggability Assessment, Jinan University, International Cooperative
Laboratory of Traditional Chinese Medicine Modernization and Innovative Drug Development of Chinese Ministry
of Education, Institute of New Drug Research, Jinan University College of Pharmacy, Guangzhou, China
- State Key Laboratory of Bioactive Molecules and Druggability Assessment, Jinan University, International Cooperative
Laboratory of Traditional Chinese Medicine Modernization and Innovative Drug Development of Chinese Ministry
of Education, Institute of New Drug Research, Jinan University College of Pharmacy, Guangzhou, China
Keywords:
iTRAQ-based proteomics analysis, doxorubicin, cardiotoxicity, neural cell adhesion molecule 1, DT-010, Sirt1/PGC-1α/HO-1.
Abstract:
Background: Doxorubicin (DOX) causes lethal cardiotoxicity, which limits its clinical
utility. The molecular mechanisms and effective strategies to combat its cardiotoxicity
need further exploration. DT-010, a novel conjugate of danshensu (DSS) and tetramethylpyrazine(
TMP), is considered a promising candidate for treating DOX-induced cardiotoxicity. In
this study, we aimed to investigate the underlying molecular mechanisms of DOX-induced
cardiotoxicity and the cardioprotective effects of DT-010.
Methods: Isobaric tags for relative and absolute quantitation (iTRAQ) in proteomics analysis
was employed to analyze the differentially expressed proteins in DOX-injuried hearts. Gene
ontology (GO) enrichment analysis and the Kyoto Encyclopedia of Genes and Genomes
(KEGG) pathway analysis were carried out to evaluated the potential mechanisms of DOXinduced
cardiotoxicity. The effects of NCAM1 on DOX-induced cardiotoxicity in H9c2 cells,
as well as the cardioprotection of DT-010 were assessed through NACM1siRNA transfection,
cell viability assay, cell apoptosis staining, reactive oxygen species measurement, and western
blotting.
Results: Proteomics analysis revealed that several signaling pathways, including the tricarboxylic
acid (TCA) cycle and oxidative phosphorylation, were involved in DOX-induced cardiotoxicity.
NCAM1 is one of the significantly changed proteins. DT-010 treatment regulated
NCAM1 protein expression. Silencing NCAM1 in DOX-treated H9c2 cells decreased cell viability,
increased cell apoptosis and reactive oxygen species (ROS) generation, and attenuated
the cardioprotective effects of DT-010. Furthermore, NCAM1 knockdown promoted p38 activation
and inhibited the expressions of peroxisome proliferator-activated receptor gamma coactivator-
1 alpha (PGC-1α) and heme oxygenase-1 (HO-1) in DOX-treated cells.
Conclusion: These findings indicate a definite role of NCAM1 in DOX-induced cardiotoxicity
and DT-010-exerted cardioprotection, which is mediated through the p38 and Sirt1/PGC-
1α/HO-1 pathway.
