Objective To explore the function of brain-lipid binding protein (BLBP) in cultured rat astrocytes in vitro. Methods After culturing rat astrocytes, BLBP small interference RNA (siRNA) was used to silence the expression of endogenous BLBP. Real-time PCR and Western blotting were applied to detect the level of BLBP and c-Myc; the cells viability was detected by using CCK-8 regent, the proliferated cells were labeled by Ki67, and the cells cycle was investigated by flow cytometry. Results The expressions of BLBP and c-Myc were significantly decreased after treatment of BLBP siRNA. BLBP gene silencing suppressed the cells viability and decreased the proportion of Ki67-positive cells. The result of flow cytometry showed that the insufficient expression of BLBP reduced the cells number of S phase. Conclusion The gene silencing of BLBP inhibits the cells proliferation of astrocytes in vitro. BLBP may regulate the proliferation process of astrocytes.