Chick embryo partial ischemia model: a new approach to study ischemia ex vivo.
BACKGROUND
Ischemia is a pathophysiological condition due to blockade in blood supply to a specific tissue thus damaging the physiological activity of the tissue. Different in vivo models are presently available to study ischemia in heart and other tissues. However, no ex vivo ischemia model has been available to date for routine ischemia research and for faster screening of anti-ischemia drugs. In the present study, we took the opportunity to develop an ex vivo model of partial ischemia using the vascular bed of 4(th) day incubated chick embryo.


RESULTS
Ischemia was created in chick embryo by ligating the right vitelline artery using sterile surgical suture. Hypoxia inducible factor- 1 alpha (HIF-1alpha), creatine phospho kinase-MB and reactive oxygen species in animal tissues and cells were measured to confirm ischemia in chick embryo. Additionally, ranolazine, N-acetyl cysteine and trimetazidine were administered as an anti-ischemic drug to validate the present model. Results from the present study depicted that blocking blood flow elevates HIF-1alpha, lipid peroxidation, peroxynitrite level in ischemic vessels while ranolazine administration partially attenuates ischemia driven HIF-1alpha expression. Endothelial cell incubated on ischemic blood vessels elucidated a higher level of HIF-1alpha expression with time while ranolazine treatment reduced HIF-1alpha in ischemic cells. Incubation of caprine heart strip on chick embryo ischemia model depicted an elevated creatine phospho kinase-MB activity under ischemic condition while histology of the treated heart sections evoked edema and disruption of myofibril structures.


CONCLUSIONS
The present study concluded that chick embryo partial ischemia model can be used as a novel ex vivo model of ischemia. Therefore, the present model can be used parallel with the known in vivo ischemia models in understanding the mechanistic insight of ischemia development and in evaluating the activity of anti-ischemic drug.
Majumder
Syamantak
S
Vascular Biology Lab, AU-KBC Research Centre, Anna University, Chennai, India.
Ilayaraja
M
M
Seerapu
Himabindu Reddy
HR
Sinha
Swaraj
S
Siamwala
Jamila H
JH
Chatterjee
Suvro
S
D000083:Acetanilides / Q000494:pharmacology; D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D001808:Blood Vessels / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002642:Chick Embryo; D052279:Creatine Kinase, MB Form / Q000378:metabolism; D004195:Disease Models, Animal; D004487:Edema / Q000150:complications / Q000473:pathology; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D005978:Glutathione / Q000378:metabolism; D006041:Goats; D006262:Health; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000378:metabolism; D007511:Ischemia / Q000473:pathology*; D009206:Myocardium / Q000201:enzymology / Q000473:pathology; D009389:Neovascularization, Pathologic / Q000378:metabolism; D009928:Organ Specificity / Q000187:drug effects; D010879:Piperazines / Q000494:pharmacology; D000069458:Ranolazine; D017382:Reactive Oxygen Species / Q000378:metabolism; D013997:Time Factors; D014656:Vascular Surgical Procedures / Q000379:methods*
D000083:Acetanilides; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D010879:Piperazines; D017382:Reactive Oxygen Species; D000069458:Ranolazine; D052279:Creatine Kinase, MB Form; D005978:Glutathione