WANG Jingbo , HUI Yannian , HAN Quanhong , et al
  • Department of Ophthalmology，Xijing Hospital，Fourth Military Medical University，Xi′;
  • an 710032,China;
Pigment epithelium of eye／drug effects; Tumor necrosis factor; Lipopolysaccharide/analysis; Protein kinase／analysis
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Objective To investigate the effects of cytokines on the expression of syndecan-1 in cultured human retinal pigment epithelial (RPE) cells and the signal transduction pathway. Methods Reverse transcription polymerase chain reaction and immunofluorescence staining were used to detect the expression of syndecan-1 mRNA and protein in normal RPE cells. The expression of syndecan-1 in RPE cells stimulated by different cytokines was detected and quantitatively analyzed by image process of immunofluorescence. The stimulation included 7 and 35 ng/ml tumor necrosis factor (TNF)- alpha; for 24 hours, 1 and 6  mu;g/ml lipopolysaccharide (LPS) for 11 hours, 7 ng/ml TNF- alpha; for 0 to 24 hours (once per 2 hours, and 13 times in total), and 30% supernatant of monocyte/macrophage strain (THP-1 cells) for 3, 14 and 43 hours. The effect of 30% supernatant of THP-1 cells was assayed after pretreated by PD098059［the specific inhibitor of extracellular signal regulated kinase(ERK) 1/2］for 2 hours. After exposed to 30% supernatant of THP-1 cells for 3 hours and treated by 0.25% trypsin for 5 minutes, RPE cells attaching was evaluated by methyl thiazolyl tetrazolium assay. Results In normal human RPE cells, expressions of syndecan-1 mRNA and protein were detected, and b syndecan-1 positive yellowish green fluorescence was found in the cell membrane and cytoplasm while light green fluorescence was in the nucleus. As the concentration and stimulated time of TNF- alpha; or LPS increased, the fluorescence intensity decreased（P lt;0.01）, and after exposed to 30% supernatant of THP-1 cells, weaker fluorescence intensity was detected (P lt;0.001). Pretreatment with 50  mu;mol/L PD098059 for 2 hours partly inhibited the effect of THP-1 cells supernatant. After exposed to 30% supernatant of THP-1 cells for 3 hours, the number of attached cells decreased compared with the controls（P lt;0.05）. Conclusions TNF- alpha; and LPS down-regulate the expression of syndecan-1 in cultured human RPE cells. The supernatant of THP-1 cells down-regulates the expression of syndecan-1 and lessens the cells attaching, which is at least mediated by ERK 1/2 pathway. (Chin J Ocul Fundus Dis, 2006, 22: 113-116)

Citation： WANG Jingbo,HUI Yannian,HAN Quanhong,et al. Effects of cytokines on the expression of syndecan-1 in cultured human retinal pigment epithelial cells. Chinese Journal of Ocular Fundus Diseases, 2006, 22(2): 113-116. doi: Copy