Cell Stress and Chaperones

, Volume 23, Issue 2, pp 213–222 | Cite as

Sardine oil loaded vanillic acid grafted chitosan microparticles, a new functional food ingredient: attenuates myocardial oxidative stress and apoptosis in cardiomyoblast cell lines (H9c2)

  • K. V. Vishnu
  • K. K. Ajeesh Kumar
  • Niladri S. ChatterjeeEmail author
  • R. G. K. Lekshmi
  • P. R. Sreerekha
  • Suseela Mathew
  • C. N. Ravishankar
Original Paper


Fish oil has been widely recognized as an excellent dietary source of polyunsaturated n-3 fatty acids such as EPA and DHA. However, it can undergo oxidation easily resulting in the formation of toxic off flavor compounds such as hydroperoxides. These compounds adversely affect the nutritional quality and may induce several stress reactions in body. To solve this problem, a new antioxidant bio-material, vanillic acid-grafted chitosan (Va-g-Ch), was synthesized and used as a wall material for microencapsulation of fish oil. The sardine oil loaded Va-g-Ch microparticles could be a potential functional food ingredient considering the numerous health benefits of fish oil, chitosan, and vanillic acid. The current study aimed to investigate the possible protective effect of sardine oil-loaded Va-g-Ch microparticles against doxorubicin-induced cardiotoxicity and the underlying mechanisms. In vitro cytotoxicity evaluation was conducted using H9c2 cardiomyocytes. MTT assay revealed that effective cytoprotective effect was induced by a sample concentration of 12.5 μg/mL. Results of apoptosis by double fluorescent staining with acridine orange/ethidium bromide and caspase-3 evaluation by ELISA substantiated the above findings. Further, flow cytometric determination of membrane potential, relative expression of NF-κB by PCR, and ROS determination using DCFH-DA also confirmed the protective effect of encapsulated sardine oil against doxorubicin-induced cardiotoxicity. NF-κB expression was down-regulated nearly by 50% on cells treated with encapsulated sardine oil. Altogether, the results revealed that sardine oil-loaded Va-g-Ch microparticles demonstrated potential cell protection against doxorubicin-induced oxidative stress


Sardine oil Cardiotoxicity Doxorubicin Microencapsulated fish oil ROS scavenging NF-κB 


Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflicts of interest.

Supplementary material

12192_2017_834_MOESM1_ESM.docx (1 mb)
ESM 1 (DOCX 1037 kb)


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Copyright information

© Cell Stress Society International 2017

Authors and Affiliations

  • K. V. Vishnu
    • 1
  • K. K. Ajeesh Kumar
    • 1
  • Niladri S. Chatterjee
    • 1
    Email author
  • R. G. K. Lekshmi
    • 1
  • P. R. Sreerekha
    • 1
  • Suseela Mathew
    • 1
  • C. N. Ravishankar
    • 1
  1. 1.ICAR-Central Institute of Fisheries Technology (CIFT)KeralaIndia

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