Abstract
Abstract: :
Purpose: This study evaluated the effects of glucose on proliferation, migration, and proteinase expressions in human retinal pigment epithelial cells to discover the cause of diabetic retinal complications. Methods: Human retinal pigment epithelial cells (CRL 2302) were cultured in media containing 5.5mM, 11.0mM, and 16.5mM D–glucose to simulate diabetic condition. The present study performed a MTT analysis for the proliferation assay and counted a number of cells for the migration assay. In addition, the study conducted western blotting for the expression of protein of matrix metalloproteinase(MMP)–2, –9, tissue inhibitor of matrix metalloproteinase(TIMP)–1, –2 , and RT–PCR for the expression of mRNA of MMP–2,9, and TIMP–1, –2. Results: The findings of the MTT and cell–count indicated that raising concentration of glucose resulted in a significant decrease in cellular proliferation and migration. The results of the western blotting analysis illuminated that raising concentration of glucose significantly increased the expression of MMP–2 and –9 but significantly decreased the expression of TIMP–1 and –2. Moreover, the RT–PCR results indicated significant increases in the expression of mRNA of MMP–2 and –9 as well as in the expressions of mRNA of TIMP–1 and –2 by raising glucose concentration. Conclusions: Higher levels of glucose showed significant effect on the cellular proliferation, migration, and expression of MMP and TIMP in human retinal pigment epithelial cells. This study provides fundamentals in later work on the mechanism of retinal complication in diabetic patients.
Keywords: retinal pigment epithelium • enzymes/enzyme inhibitors • diabetes