Abstract: Substances of abuse, such as opiates, and astroglial-derived proinflammatory cytokines, such as interleukin (IL)-1beta, likely contribute to the neuroinflammatory and neurodegenerative processes observed in NeuroAIDS in injection drug users. Furthermore, uncontrolled synthesis and activation of complement component C3 in the brain can also lead to inflammation and neurodegeneration. We hypothesized that morphine may alter regulation of the C3 gene by IL-1beta in astrocytes. Our studies demonstrate that IL-1beta induces C3 promoter activity in a CAAT/enhancer-binding protein (C/EBP)-dependent manner. Inhibition of IL-1beta mediated C3 promoter activation by the dominant negative mutant of p38-alpha mitogen-activated protein kinase suggests that IL-1beta induces C3 expression through the activation of C/EBP. Morphine (0.01 microM) in combination with IL-1beta further induced C3 promoter activity. Similarly, the C/EBP-beta isoform liver activating protein and C/EBP-delta-induced C3 promoter activity were upregulated by morphine and IL-1beta. Taken together, this study illustrates that morphine modulates IL-1beta-mediated C3 expression in astrocytic cells.