Abstract
Purpose: :
To compare differentially expressed genes concerned with glucose between normal and diabetic keratocytes stimulated with and without interleukin-1α (IL-1α) and tumor necrosis factor-α (TNF-α).
Methods: :
Normal and diabetic keratocytes were primarily cultured, and treated with 20 ng/ml of IL-1α and TNF-α for 6 hours. Hybridization with cDNA from mRNA was carried out using oligonucleotide microarray. Identified genes were further evaluated by real-time PCR.
Results: :
Diabetic keratocytes showed the different gene expression of 8 down-regulated genes including App, Dgat1, Hdc and Sgk, and 5 up-regulated genes including Sirt1 and Fabp3. IL-1α treated diabetic keratocytes demonstrated 4 down-regulated genes of all 7 differentially expressed genes. Four down-regulated genes of all 5 differentially expressed genes after TNF-α treatment. Newly and commonly expressed genes containing all under-expressed 4 genes after treatment of both cytokines were Hmox1, Il-6, Npy, and Pygl. Real time PCR showed Hmox1, IL-6 and Pygl genes were significantly decreased and Sirt1 gene was significantly increased.
Conclusions: :
There were some differentially expressed genes related with glucose between normal and diabetic keratocytes. Also IL-1α and TNF-α may stimulate to differing effects of under-regulation on glucose such as Hmox1, IL-6, Npy, and Pygl.
Keywords: cornea: basic science • cornea: stroma and keratocytes • gene/expression