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Jennifer C. Howell, Eugene Chun, Hana Kim, P Michael Iuvone, Rashidul Haque; Curcumin (Diferuloylmethane) Alters H2O2-induced Expression Profiles of MicroRNAs in Human ARPE-19 Cells. Invest. Ophthalmol. Vis. Sci. 2012;53(14):1588.
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Oxidative stress (OS) has been implicated in many ocular diseases in which the RPE is considered a primary target. Several studies have shown that miRNAs play roles in a broad range of biological processes, including AMD, retinitis pigmentosa, retinoblastoma, and diabetic retinopathy. Alterations in miRNA expression may occur following exposure to stress-inducing agents, including H2O2. This study has attempted to unravel whether curcumin, a polyphenolic compound with many biological activities, alters H2O2-induced expression profiles of miRNAs in ARPE-19 cells.
The effect of curcumin on the H2O2-induced miRNA expression profile was assessed in ARPE-19 cells, which were treated with curcumin (20 µM) for 6 h prior to H2O2 (200 µM) for 18 h. Control cells with or without H2O2 and curcumin treatments were also maintained. miRNA expression profiling was carried out through real-time PCR array (Qiagen). The array contains a panel of 88 miRNAs, plus four housekeeping genes. Also, the effect of curcumin on the expression levels of selected antioxidant genes and proteins was tested by qRT-PCR and western blotting, respectively.
Given individually, curcumin and H2O2 significantly downregulated 20 and 18 miRNAs, and upregulated 6 and 29 miRNAs (p<0.05 compared with controls; fold change >2.0), respectively. Furthermore, curcumin pretreatment in cells exposed to H2O2 significantly reduced the expression of 17 H2O2-induced miRNAs. The members of the miR-30 family that are putative regulators of the antioxidant defense system were found to be reduced by curcumin. As determined by qRT-PCR and western blotting, curcumin increased the expression of catalase, GPx1, and GPx4 at the mRNA and protein levels, respectively.
The results demonstrated that curcumin can modulate the expression of H2O2-induced miRNAs that are putative regulators of the antioxidant defense system, as well as other genes that have been reported to be linked to ocular diseases.
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