Oxidative stress is implicated in functional recovery after brain damage caused by intracerebral hemorrhage (ICH). Histone deacetylase 6 (HDAC6) plays an important role in initiation of oxidative stress. However, studies have not explored the role and mechanism of HDAC6 in mediating oxidative stress in ICH. Our study revealed HDAC6 knockout mice are resistant to ICH-induced oxidative stress and the resultant neuronal apoptosis, as analyses performed through MDA and NADPH/NADP+ assays, ROS detection and determination of expression levels apoptosis-related protein through western blot. Further mechanistic studies showed that HDAC6 binds to Malate dehydrogenase 1 (MDH1) and mediates MDH1 acetylation on lysine residues at position 121 and 298. ICH associated damaging agents reduce the level of MDH1 acetylation by promoting the interplay between HDAC6 and MDH1. Whereas, acetylation of MDH1 increases when HDAC6 is inhibited. Functional verification of acetylation MDH1 demonstrates that non-acetylation mutant MDH1, rather than its acetylation resistant mutant, protects neuron from oxidative injury. In addition, HDAC6 inhibition failed to mediate brain damage alleviation when MDH1 was knockdown. Overall, the findings of the present study indicated the protect effect of HDAC6 inhibition resistance to brain damage during ICH requires MDH1 acetylation.