Production of Nerve Growth Factor in Primary Cultured Astrocytes under Hypoxia/Reoxygenation

Objective: To investigate the secretion of nerve growth factor(NGF) in primary cultured astrocytes under hypoxia/reoxygenation(H/R). Methods: Primary cultured rats cortical astrocytes were used, which were randomly assigned into groups of normal control(N group) and hypoxia/reoxygenation(H/R group). Morphological observation of the astrocytes was detected by inverted phase contrast microscopy as an index of cellular damage under hypoxia. And tyrpan blue staining was applied to detect the cell injury rate at different time after reoxygenation. The levels of NGF protein were analyzed by enzyme-linked immune sorbent assay(ELISA). And q RT-PCR was used to determine the levels of NGF m RNA. Results: The astrocytes viability of the H/R group has no significant changed after 6 h hypoxia and 48 h reoxygenation. Compared with the N group, the NGF m RNA level of the H/R group was remarkably increased after 24 h reoxygenation and 6 h hypoxia(P0.01), and the level of NGF protein secretionof the H/R group increased significantly as well after 48 h reoxygenation and 6 h hypoxia(P0.01).Conclusion: Hypoxia/reoxygenation effectively increased the expression of NGF in primary cultured astrocytes.