A new oxidative stress model, 2,2-azobis(2-amidinopropane) dihydrochloride induces cardiovascular damages in chicken embryo

PLoS One. 2013;8(3):e57732. doi: 10.1371/journal.pone.0057732. Epub 2013 Mar 1.

Abstract

It is now well established that the developing embryo is very sensitive to oxidative stress, which is a contributing factor to pregnancy-related disorders. However, little is known about the effects of reactive oxygen species (ROS) on the embryonic cardiovascular system due to a lack of appropriate ROS control method in the placenta. In this study, a small molecule called 2,2-azobis(2-amidinopropane) dihydrochloride (AAPH), a free radicals generator, was used to study the effects of oxidative stress on the cardiovascular system during chick embryo development. When nine-day-old (stage HH 35) chick embryos were treated with different concentrations of AAPH inside the air chamber, it was established that the LD50 value for AAPH was 10 µmol/egg. At this concentration, AAPH was found to significantly reduce the density of blood vessel plexus that was developed in the chorioallantoic membrane (CAM) of HH 35 chick embryos. Impacts of AAPH on younger embryos were also examined and discovered that it inhibited the development of vascular plexus on yolk sac in HH 18 embryos. AAPH also dramatically repressed the development of blood islands in HH 3+ embryos. These results implied that AAPH-induced oxidative stress could impair the whole developmental processes associated with vasculogenesis and angiogenesis. Furthermore, we observed heart enlargement in the HH 40 embryo following AAPH treatment, where the left ventricle and interventricular septum were found to be thickened in a dose-dependent manner due to myocardiac cell hypertrophy. In conclusion, oxidative stress, induced by AAPH, could lead to damage of the cardiovascular system in the developing chick embryo. The current study also provided a new developmental model, as an alternative for animal and cell models, for testing small molecules and drugs that have anti-oxidative activities.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amidines / toxicity*
  • Animals
  • Cardiomegaly / chemically induced
  • Cardiomegaly / pathology*
  • Cardiovascular System / drug effects*
  • Cardiovascular System / pathology
  • Chick Embryo
  • Chorioallantoic Membrane / abnormalities
  • Chorioallantoic Membrane / blood supply*
  • Chorioallantoic Membrane / drug effects*
  • Lethal Dose 50
  • Models, Biological
  • Neovascularization, Physiologic
  • Oxidants / toxicity*
  • Oxidative Stress
  • Reactive Oxygen Species / agonists
  • Reactive Oxygen Species / metabolism
  • Ventricular Remodeling / drug effects

Substances

  • Amidines
  • Oxidants
  • Reactive Oxygen Species
  • 2,2'-azobis(2-amidinopropane)

Grants and funding

This work was supported by the grant from National Natural Science Foundation of China (30971493, 31071054 & 81102485) and BRAND’S Centre for Health and Nutritional Sciences in Singapore. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.