Abstract
The gap junction intercellular communication (GJIC) is considered to play an important role in intercellular communications, and hyperglycemia might cause gap junction degeneration. Alpha-lipoic acid (ALA) was commonly used for protection against diabetes and diabetic retinopathy, despite that whether it can ameliorate damages to retinal pigment epithelium (ARPE-19) under hyperglycemic stress remains unknown. In this study, ARPE-19 cells were incubated with 17.5, 25, 33, 50 mM glucose, and 100, 200, 500 μM alpha-lipoic acid at 37°C,5% CO2 for 8 days. Cell viability was determined by the Tryphan blue assay. The expression levels of Connexin 43 (Cx43), ZO-1, Occludin, RPE 65 were determined by Western blot analysis. The integrity gap junction was examined under immunofluorescence microscopy. Gap junction intercellular communication (GJIC) was determined by the scrape-load/dye transfer assay. The results showed that the size and shape of ARPE-19 cells were changed and the cells number were decreased under high glucose pressure and alpha-lipoic acid supplement, and alpha-lipoic acid could not restore gap junction proteins such as CX 43, ZO-1, Occludins’ expression, visual cycle associated protein RPE 65 had the same result, but their GJIC permeability was increased. In summary, alpha-lipoic acid could not help to improve ARPE-19 cells’ junction protein under hyperglycemia pressure and suggested that we should use alpha-lipoic acid in diary day to prevent diabetes happened instead of under diabetes condition.
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