Activated astrocytes have been implicated in the deleterious cascade of events immediately following traumatic brain injury (TBI). In this study, primary astrocytes were activated by a well-characterized stretch injury model of TBI. Nitrite production, an outcome measure for activation in astrocytes, increased significantly after injury. Using the cell-penetrating peptide (CPP) TAT, delivery of a mock-therapeutic, green fluorescent protein (GFP), was enhanced after mechanical injury of primary astrocytes. GAG production also increased in injured astrocytes, suggesting a mechanism for enhanced delivery, confirming findings published in an earlier study. These results highlight the ability of TAT to target activated astrocytes via increased GAG content, which could lead to potential TAT-based targeted therapeutics for TBI.