Evaluating the Effects of CDK Inhibitors in Ischemia–Reperfusion Injury Models
- 1.6k Downloads
CDK inhibitors have been used to induce protection in various experimental models. Kidney ischemia–reperfusion (I/R) is a form of acute kidney injury resulting in a cascade of cellular events prompting rapid cellular damage and suppression of kidney function. I/R injury, an inevitable impairment during renal transplant surgery, remains one of the major causes of acute kidney injury and represents the most prominent factor leading to delayed graft function after transplantation. Understanding the molecular events responsible for tubule damage and recovery would help to develop new strategies for organ preservation. This chapter describes procedures to study the effect of CDK inhibitors in the cellular I/R model developed from an epithelial cell line deriving from pig kidney proximal tubule cells (LLC–PK1). We briefly describe methods for determining the protective effect of CDK inhibitors such as activation of caspase 3/7, western blot analysis, gene silencing, and immunoprecipitation.
Key wordsCDK inhibitors Kidney injury Ischemia–reperfusion Renal preservation
- 2.Iyirhiaro GO, Brust TB, Rashidian J, Galehdar Z, Osman A, Phillips M, Slack RS, Macvicar BA, Park DS (2008) Delayed combinatorial treatment with flavopiridol and minocycline provides longer term protection for neuronal soma but not dendrites following global ischemia. J Neurochem 105(3):703–713PubMedCrossRefGoogle Scholar
- 12.Canela N, Orzaez M, Fucho R, Mateo F, Gutierrez R, Pineda-Lucena A, Bachs O, Perez-Paya E (2006) Identification of an hexapeptide that binds to a surface pocket in cyclin A and inhibits the catalytic activity of the complex cyclin-dependent kinase 2-cyclin A. J Biol Chem 281(47):35942–35953PubMedCrossRefGoogle Scholar
- 14.Taniguchi Y, Pippin JW, Hagmann H, Krofft RD, Chang AM, Zhang J, Terada Y, Brinkkoetter P, Shankland SJ (2012) Both cyclin I and p35 are required for maximal survival benefit of cyclin-dependent kinase 5 in kidney podocytes. Am J Physiol Renal Physiol 302(9):F1161–F1171PubMedCentralPubMedCrossRefGoogle Scholar