Glial cells are mediating the main activation of the central nervous system (CNS), being astrocytes the mayor glial cells in the brain. Glial activation may result beneficial since it could promote tissue repair and pathogen elimination. However, excessive glial activation mechanism can also have do harm to the tissue. β-1,4-Galactosyltransferase I (β-1,4-GalT-I) is a key inflammatory mediator that participates in the initiation and maintenance of inflammatory reaction in some diseases. Moreover, CDK11(p58) has been reported to be associated with β-1,4-GalT-I. We have found that CDK11(p58) and β-1,4-GalT-I are induced in lipopolysaccharide (LPS)-challenged rat primary astrocytes in a affinis dose- and time-dependent manner. CDK11(p58) regulates the expression of β-1,4-GalT-I by interacting with it. After the knockdown of CDK11(p58) expression, the expression of β-1,4-GalT-I decreases, and astrocyte activation downregulates. Inversely, the expression of β-1,4-GalT-I increases, and astrocyte activation enhances due to the overexpression of CDK11(p58). Knockdown of β-1,4-GalT-I reduces the activation potentiation caused by the overexpression of CDK11(p58), illustrating the function of CDK11(p58) to promote astrocyte activation depends on β-1,4-GalT-I. The interaction between CDK11(p58) and β-1,4-GalT-I to upregulate astrocyte activation is related to activating p38 and JNK pathways. These findings indicated that the functional interaction between CDK11(p58) and β-1,4-GalT-I may play an important role during astrocyte activation after LPS administration.