Effect Of Hypoxia On The Expressions Of VEGF Or BDNF MRNA And Protein In Cerebral Cortex Cells Of SD Rats

Objective:To observe the effect on the expressions of vascular endothelial growth factor (VEGF) mRNA and protein、brain derived nerve growth factor (BDNF) mRNA and protein in cerebral cortex cells under the environment of different oxygen concentration and time treatment.Methods:The primary cerebral cortex cells of SD rats were designed into two hypoxia group and two normoxia control groups after5days. Then, the cells of hypoxia groups were treated by3hours (3h) and6hours (6h) under the environment of1%and4%oxygen concentration respectively in hypoxia workstation, while the corresponding control groups remained in HHCP continuely. The ELISA method was used to detect VEGF protein content in cultural supernatants, and the level of intracellular VEGF mRNA was assayed by RT-PCR. protein expression and identification The phenotypc of cortex cells and VEGF or BDNF protein expression were identified by immunohistochemical double staining. The comparison of two groups use t test.Each experiment repeats3times independently.Results:1. Compared to the control groups, the level of VEGF mRNA and protein of3h groups had no significant difference under the environment of1%and4%oxygen concentration, but had increase trend, while the level of6h groups were up-regulated obviously, and the increases in1%oxygen concentration environment were greater.2. The level of BDNF mRNA and protein of3h groups had no heterogeneity in either1%and4%oxygen concentration from that in control group, but had a trend of rising, while the level of6h groups were increased significantly. Among the rest, the protein expressions of1%oxygen concentration increased more, and the mRNA of4%oxygen concentration increased greater.3. The cortex cells grow in colonies inordinately after processing of hypoxia.4.The protein fluorescence intensity of immunopositive Anti-VEGF or Anti-BDNF enhanced after hypoxia. Moreover, the overlap ratio of immunopositive GFAP astrocytes and immunopositive Anti-VEGF or Anti-BDNF cover78%and70%in all cells. Conclusion:1. The expressions of VEGF or BDNF mRNA and protein in cerebral cortex cells can be induced by hypoxia, and the response of cerebral cortex cells has related with the concentration and treatment time of hypoxia.2.Whether or not the beginning of cell inoculating density was suitable, which could be accounted for the phenomenon of cells grow in cluster.3. The expressions of VEGF and BDNF induced by hypoxia mainly derived from neurons and astrocytes, but more from the latter.