Figure 6.

TMP attenuates H₂O₂-induced PRGC apoptosis via miR-182/mitochondrial apoptotic pathway. Cells were transfected with miR-182 inhibitor or inhibitor NC prior to treatment with TMP for 24 h and then exposed to H₂O₂. (A) Western blot analysis was conducted to determine the expression of apoptosis-related proteins (BNIP3, Bcl-2, cle-caspase-3, Bax and cle-PARP and Bcl-2). (B) Protein levels of cyto c in mitochondria and cytosol was measured using western blot analysis. β-actin and Cox IV were used as loading controls for the cytosolic and mitochondrial fractions, respectively. (C) Mitochondrial membrane potential levels in the treated PRGCs were analyzed using the JC-1 fluorescent probe. Data are presented as the mean of three replicates ± standard deviation. ^*P<0.05, ^**P<0.01 vs. Control group, ^##P<0.01 vs. H₂O₂ group, ^$$P<0.01 vs. H₂O₂ + TMP group. Bcl-2, B-cell lymphoma 2; Bax, Bcl-2-assoiated X protein; BNIP3, Bcl-2 interacting protein 3; cle, cleaved; Cox, IV, Complex IV; Cyto c, cytochrome c; miR, microRNA; PARP, poly(ADP-ribose)polymerase; PRGCs, primary retinal ganglion cells; TMP, tetramethylpyrazine.