| Objective:To investigate ①if the gene expressions associated with 5-HTT, NET, BDNF, and IDO were modulated by CORT in primarily cultured hyppocampal neurons; ②if the gene expressions associated with 5-HTT, NET, BDNF, and IDO were also modulated by vaproate.Methods:1. Primarily cultured fetal hippocampal neurons were from SD rats with 18d-pregnancy. Immunohistochemical method was used to identify the purity of neurons.2. MTT assay was used to determine concentration-effect and time-effect of CORT on the cultured hippocampal neurons.3. RT-PCR and Western-blot were used to determine whether CORT and vaproate affected on the expressions of 5-HTT, NET, BDNF, IDO mRNA and protein through CORT receptor in the cultued hippocampal neurons, respectively.Results:1. The purity of primarily cultured hippocampal neurons was more than 90% with vivid appearance and function.2. MTT assay showed that cell viability was decreased in time-and concentration-dependent manner after co-incubation of hippocampal neurons with CORT; a significant decrease of cell viability was observed when CORT was used at 5μmol/L for 24h or 1μmol/L for 48h. There was no significant change of cell viability when the concentration of CORT was lower than 1 μmol/L3. RT-PCR and Western blot showed that CORT significantly decreased the expression of BDNF mRNA and protein but increased the expression of IDO mRNA and protein, from hippocampal neurons, which could be reversed by mifepristone rather than sodium valporate. However, CORT failed to change mRNA and protein expressions of monoamine transporters in hippocampal neurons.Conclusion:1. Corticosterone can down-regulate the expression of BDNF and up-regulate the expression of IDO through acting on corticosterone receptor, but can not change the expression of monoamine transporters in hippocampal neurons.2. Vaproate can not reverse the modulating effects of corticosterone on the expressions of both BDNF and IDO in hippocampal neurons... |
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