Renal
ischemia reperfusion injury contributes patho-physiological imbalance of
acute renal failure that comprises of generation of
reactive oxygen species,
nitric oxide and
peroxynitrite and
inflammation involving
cytokine/adhesion molecule cascade, finally leads to cell death.
Oxygen deprival associated with
ischemia that in turn lead to decline
ATP production is the characteristic feature usually addressed in the development of in vitro cell based ischemic model. In order to create
oxygen deficit in the cell lines different approaches like chemical induction, enzymatic induction and anaerobic chamber models are widely used. However efficiencies of these models were varied and the present study was aimed to compare the suitability of these models in creating in vitro
ischemia reperfusion in cell culture. In the chemical induced method we used different concentrations of
rotenone,
antimycin and
sodium azide to inhibit electron transport chain and thereby reduced the
ATP production, measured indirectly by cell viability assay. Among the chemical induced model,
antimycin mediated cell injury was more reliable for
ischemia reperfusion study. In the enzymatic model, comprises of
glucose oxidase (3mM/s) and
catalase (998 s(-1)
at 10:1 ratio) was used and found to be best among the three approaches as it can create injury in short experimental time and are reproducible. However anaerobic chamber method was not suitable for
ischemia reperfusion study as it need more time to induce significant cell injury.