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Changes in gene expression by deregulated microRNAs (miRNAs) are involved in the process of central nervous system neurodegeneration. Here, we report that mir\u2010145, a miRNA enriched in rat spinal neurons and astrocytes, was downregulated at 1 week and 1 month after SCI. Our <jats:italic>in vitro<\/jats:italic> studies using astrocytes prepared from neonatal spinal cord tissues indicated that potent inflammagen lipopolysaccharide downregulated mir\u2010145 expression in astrocytes, suggesting that SCI\u2010triggered inflammatory signaling pathways could play the inhibitory role in astrocytic mir\u2010145 expression. To induce overexpression of mir\u2010145 in astrocytes at the spinal cord lesion site, we developed a lentivirus\u2010mediated pre\u2010miRNA delivery system using the promoter of glial fibrillary acidic protein (GFAP), an astrocyte\u2010specific intermediate filament. The results indicated that astrocyte\u2010specific overexpression of mir\u2010145 reduced astrocytic cell density at the lesion border of the injured spinal cord. In parallel, overexpression of mir\u2010145 reduced the size of astrocytes and the number of related cell processes, as well as cell proliferation and migration. Through a luciferase reporter system, we found that GFAP and c\u2010myc were the two potential targets of mir\u2010145 in astrocytes. Together, the findings demonstrate the novel role of mir\u2010145 in the regulation of astrocytic dynamics, and reveal that the downregulation of mir\u2010145 in astrocytes is a critical factor inducing astrogliosis after SCI. 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