PRDX2 silencing alleviates reactive hyperplasia of Müller glial cells in diabetic retinopathy by inhibiting the RhoA/ROCK signaling pathway

Figure 2 PRDX2 is upregulated in ocular tissues of patients with diabetic retinopathy and high-glucose-induced Müller cells. A: Vitreous PRDX2 protein in proliferative diabetic retinopathy (DR; n = 9) vs non-diabetic controls (n = 6; ELISA); B: Retinal PRDX2 expression in six DR patients vs five healthy controls (GSE53257); C: PRDX2 gene expression in the single cell type clusters in the human retina from the Human Protein Atlas; D-L: RMC-1 cells were treated with high glucose (25 mmol/L) or normal glucose (5.5 mmol/L) for 24 hours or 48 hours to mimic diabetes-related stress; Apoptosis by Annexin V/PI flow cytometry. The bar histogram shows the quantitative results of flow cytometry. Living cell populations are clustered in the Q4 quadrant; apoptotic or necrotic cells are in the Q2 and Q3 quadrants (D and E). Cell viability of rMC-1 cells by CCK8 assay (F). Migration of rMC-1 cells by wound-healing assay (G and H; magnification 100 ×; scale bars: 100 μm). The bar histogram shows the percentage of the cell migration area. Photomicrographs (40 × and 100 ×) show the invasive ability of rMC-1 cells as measured by the Transwell chamber assay (I and J). Lactate content of rMC-1 cells (K). mRNA expression of PRDX2 and PRDX6 measured by quantitative PCR (L). All values are presented as means ± SEM. Each experiment was independently repeated 3-5 times. P value for trends assessed using the t-test or ordinary one-way analysis of variance. ^aP < 0.05; ^bP < 0.01; ^cP < 0.001; ^dP < 0.0001. PDR: Proliferative diabetic retinopathy; DR: Diabetic retinopathy; NG: Normal glucose; HG: High glucose.