Glutathione (GSH) is an important antioxidants that plays a critical role in neuroprotection. GSH depletion in neurons induces oxidative stress and thereby promotes neuronal damage, which in turn is regarded as a hallmark of the early stage of neurodegenerative diseases. The neuronal GSH level is mainly regulated by excitatory amino acid carrier 1 (EAAC1) and its inhibitor, glutamate transporter-associated protein 3-18 (GTRAP3-18). In this study, we found that GTRAP3-18 level was increased by the up-regulation of the microRNA miR-96-5p, which has been reported to decrease EAAC1 level in our previous study. Since the 3’-UTR region of GTRAP3-18 lacks the consensus sequence for miR-96-5p, an unidentified protein should intermediately regulate GTRAP3-18 expression by miR-96-5p. Here we discovered that neuro-oncological ventral antigen 1 (NOVA1) is an intermediate protein for GTRAP3-18 expression via miR-96-5p. Moreover, we show that the administration of a miR-96-5p-inhibiting nucleic acid to living mice by a drug delivery system decreased the level of GTRAP3-18 via NOVA1 and increased the levels of EAAC1 and GSH in mouse brain. These findings suggest that the delivery of a miR-96-5p inhibitor to the brain would efficiently increase the neuroprotective activity by increasing GSH levels via EAAC1, GTRAP3-18 and NOVA1.