The Effects Of Interferon-alpha On Expressions Of Genes Associated With Depression In Human Neuroblastoma SH-sy5y Cells

BackgroundIFN-α as an antiviral,anticancer drugs widely used in clinical practice,but the long course application prone to depression and adverse reactions,a higher incidence of disease.At present the pathogenesis of depression is uncertain,during which the central monoamine neurotransmitter system dysfunction the most important hypothesis that,because the brain monoamine neurotransmitters such as serotonin(5-HT),noradrenaline(NE)and so on with low functional level,signaling between brain cells fails,thus emerging the depression-like behavior.It has recently been reported that the expression changes of adcy3,dgka,fam46 a,igsf4a,kiaa1539,marcks,raph1,psme1 in peripheral blood are closely related to depression.Therefore,to clarify IFN-α molecular mechanisms causing depression,which will be conducive to display the biggest effectiveness of IFN-α.ObjectiveTo investigate the effects of IFN-α on human neural cells adcy3,dgka,fam46 a,igsf4a,kiaa1539,marcks,raph1,psme1 expression levels.Methods1.Cell cultures:Put the SH-sy5 y cells seeded in RPMI1640 medium containing 10%inactivated calf serum containing 25 mMHepes,2mML glutamine,100U/mL penicillin,100ug/mL streptomycin cells placed in the 37℃,5%CO2,70%-80% humidified incubator to culture and the medium was changed once every other three days.2.Drug treatment : Cultured SH-sy5 y cells were divided into two groups :Experimental group concentration of 1000 IU / ml interferon-α-2b injection,while thecontrol group with PBS buffer with drug stimulation for 16 hours.3.Real Time PCR Detection:Extract the whole cellular RNA,reverse transcribed them into cDNA,and design primers at both ends according to known human expressed sequence of genes,with cDNA as a template to amplification,and then the Real Time PCR will detect the expression of genes of adcy3,dgka,fam46 a,igsf4a,kiaa1539,marcks,raph1,psme1.4.Statistical analysis : The results were expressed as mean ± standard deviation(x ±s),with the application of one-wayanova,and independent sample to test.And it will have the statistical meaning on the condition of P <0.05.Employ the GraphPad Prism 5 software for analyzing the results and make the histogram.ResultReal-time PCR showed,adcy,dgka,fam46,marcks,raph1,psme1 gene expression level in PBS group and IFN-α group was significantly different(P <0.05),adcy3 and marcks gene expression reduced than the PBS group,dgka,fam46 a,raph1 and psme1 gene expression increase than the PBS group,and there is no significant difference in expression while igsf4 a and kiaa1539 in the two groups(P >0.05).Conclusion1.IFN-α could significantly reduce the expression of adcy3 and marcks genes in SH-sy5 y cells;but it could significantly increase the expression of dgka,fam46 a,raph1and psme1 genes.2.IFN-α had no significant effect on the expression of igsf4 a and kiaa1539 genes in SH-sy5 y cells.