Objective: To investigate the inhibition of IL-6 antisense oligonucleotide (ASON) on IL-6 expression by retinal pigment epithelium (RPE) cells on the basis of previous study.
Methods: Cultured human RPE cells was treated with IL-1 beta and IL-6 ASON. IL-6 mRNA and protein expression were detected by enzyme linked immunosorbent assay (ELISA), immunohistochemistry and in situ hybridization histochemistry (ISH).
Results: It was demonstrated that the IL-6 expression by RPE cells was dose and time dependent after the stimulation of IL-1 beta. IL-6 in the conditioned media or in the control group was 2.00 x 10(-6) g/L cells after the exposed to IL-1 beta for 8 hours. IL-6 ASON (2.00 x 10(-5) mol/L) obviously inhibited IL-6 (5.50 x 10(-7) g/L cells, t = 4.518, P < 0.01) production. Immunohistochemistry study showed dark blue staining in RPE cytoplasm after stimulation with IL-1 beta, while the cells treated with IL-6 ASON showed light staining in RPE cytoplasma. ISH displayed that there was a marked reduction in mRNA expression in IL-6 ASON treated group compared with that in the control group (t = 3.746, P < 0.01).
Conclusion: Cultured RPE cells express IL-6 protein and mRNA in dose and time dependent manners when RPE cells are stimulated with IL-1 beta. The expressions of IL-6 protein and mRNA can be significantly inhibited by IL-6 ASON in cultured human RPE cells.
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