Abstract
Purpose: :
To identify differentiate genes concerned with apoptosis that are up–regulated or down–regulated in OLETF corneal keratocyte stimulated TNF–α
Methods: :
Primary corneal keratocyte cultures of OLETF (Otsuka Long Evans Tokushima Fatty, Japan) was performed, and treated with 20 ng/ml of TNF–α for 8 hours. Hybridization was carried out using Oligonucleotide microarray. A normal keratocyte sample was labeled with Cy3, and diabetic keratocytes sample was labeled with Cy5. The confirmation of gene was performed by RT–PCR
Results: :
Compared with control, diabetic corneal keratocyte had differently several genes like clusterin, lectin galactose binding soluble, angiotensin II type–1 receptor, calpain, neural cell adhesion molecule. TNF–α treated diabetic corneal keratocyte showed down–regulated the expression of 30 genes and up–regulated 21 genes from diabetic keratocyte. The newly expressed genes concerned with TNF–α were as bone morphogenetic protein, insulin–like growth factor, tissue inhibitor of metalloproteiinase, suppressor of cytokine signaling, androgen receptor–related apoptosis–associated protein.
Conclusions: :
TNF–α may stimulate to differing effects of apoptosis on diabetic corneal keratocytes. Studies to analyze the apoptotic significance of the identified differences of diabetes are needed.
Keywords: gene/expression • apoptosis/cell death • diabetes