Figure 3.
ACh administration decreased H/R-induced mtROS levels and UPRmt via M3AChR. (A) ACh diminished H/R-induced mtROS levels in a dose-dependent manner. In the mitochondrial ROS assay, MitoSOX Red fluorescence was colocalized with that of MitoTracker Green. Scale bar, 50 μm. (B and C) ACh inhibited the expression of HSP60 and LONP1 in a dose-dependent manner. Open bar, normoxia; filled bar, H/R. (D) ACh increased the reduction in the nuclear DNA-encoded ATP5A/mitochondrial DNA-encoded MTCO1 ratio. (E) ACh treatment decreased the upregulation of HSP60 and LONP1 expression induced by H/R. The beneficial effect of ACh was abolished by the M3AChR antagonist 4-DAMP (10−6M). Open bar, normoxia; filled bar, H/R. The data expressed as mean ± SEM in each bar graph represent the average of 4 independent experiments. ***P < 0.001 vs. Con; ##P < 0.01 vs. H/R. ###P < 0.001 vs. H/R; &&P < 0.01 vs. H/R + ACh; &&&P < 0.001 vs. H/R + ACh.