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Jing Feng, Yanrong Jiang, ; Serum amyloid A supports RF/6A cells under chemical hypoxia and high glucose. Invest. Ophthalmol. Vis. Sci. 2014;55(13):2246.
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We investigated the expression of serum amyloid A (SAA) in the eyes with diabetic retinopathy. We searched for the effects of high glucose and hypoxia on the expression of SAA in RF/6A cells in vitro, and for the effects of exogenous SAA on the proliferation, migration and apoptosis of RF/6A cells.
A total of 26 eyes of 26 patients with DR, and 10 eyes of 10 patients with non-diabetic ocular disease were enrolled. Multiplex bead assay was used to examine the serum amyloid A level in aqueous humor.Localization of SAA was detected using immunofluorescence. The mRNA and protein of SAA in the RF/6A cells were measured using real-time PCR and western blot. CCK-8 assay was performed to examine the proliferative ability, transwell chamber assay and Annexin/PI staining were used to detect cell migration and apoptosis.
Significantly higher concentrations of SAA were found in the aqueous humor of DR patients than those in the aqueous humor of the control patients (p<0.01). Immunofluorescence analysis showed SAA were expressed in RF/6A cells in vitro. The expression of SAA mRNA and protein were increased induced by hypoxia and high concentration glucose (p<0.01). By CCK-8 assay, after 24h, the values were significant different with those of control groups either (p<0.01). Results of apoptosis confirmed the apoptosis rates of cells exposed to SAA for 24h were higher than those exposed to its control.
The expression of SAA was higher in DR patients. SAA was unregulated by high glucose concentration and hypoxia, which promoted cell proliferation, migration and inhibited apoptosis.
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