Fluoxetine Hydrochloride In The Regulation Of Neural Stem Cell Signaling Pathways And Downstream Gene Expression In Experimental Research

Objective To apply selective specific inhibitor(LY294002) of PI3K to incubate the rat neural stem cells to research the effect of PI3K signal transduction pathway in neural stem cells.Methods To isolate,culture the NSCs derived from E15-16 rats,carry out WTS-8 assay,BrdU andβTubulin-Ⅲimmunofluorescent staining after incubated NSCs in different concentration of LY294002.Results There was not obvious effect in the survival,proliferation and differentiation of NSCs incubated by low concentration LY294002.The viability of the NSCs decreased significantly than the control(P＜0.05),the amount of BrdU andβTubulin-Ⅲpositive cells were notably fewer than the control(P＜0.05) after they were incubated by the higher concentration LY294002.Conclusion The PI3K signal transduction pathway played a vital role in the survival,proliferation and differentiation of NSCs. Objective To apply selective specific inhibitor(PD98059) of MAPKK/MEK to incubate the rat neural stem cells to research the effect of MAPK signal transduction pathway in neural stem cells.Methods To isolate,culture the NSCs derived from E15-16 rats,carry out WTS-8 assay,BrdU andβTubulin-Ⅲimmunofluorescent staining after incubated NSCs in different concentration of PD98059.Results There was not obvious effect in the survival,proliferation and differentiation of NSCs incubated by low concentration PD98059.The viability of the NSCs decreased significantly than the control(P＜0.05 ),the amount of BrdU andβTubulin-Ⅲpositive cells were notably fewer than the control(P＜0.05) after they were incubated by the higher concentration PD98059.Conclusion The MAPK signal transduction pathway played a vital role in the survival,proliferation and differentiation of NSCs. Objective To apply the phosphatidylinositol-3-kinase(PI3-K) inhibitor(LY294002),the mitogen-activated protein kinase kinase(MEK) inhibitor(PD98059) and antidepressant drugs fluoxetine(the selective serotonin reuptake inhibitors) to incubate the rat neural stem cells to research the effect of the role of PI3K/AKT and MAPK/ERK and signaling pathways in fluoxetine treated neural stem cells.Methods To isolate,culture the NSCs derived from E15-16 rats and carried out the Western Blotting detection after respectively incubated NSCs with the phosphatidylinositol-3-kinase(PI3-K) inhibitor (LY294002),the mitogen-activated protein kinase kinase(MEK) inhibitor (PD98059) and fluoxetine.Results The fluoxetine can activated AKT,ERK1/2 protein expression of rats NSCs.Conclusion The fluoxetine can activated AKT,ERK1/2 protein expression of rats NSCs and there was cross-talk between PI3K/AKT pathway and MAPK/ERK pathway at the level of AKT-ERK1/2 in the process of fluoxetine activated AKT,ERK1/2 protein. Objective To apply the phosphatidylinositol-3-kinase(PI3-K) inhibitor(LY294002),the mitogen-activated protein kinase kinase(MEK) inhibitor(PD98059) and antidepressant drugs fluoxetine(the selective serotonin reuptake inhibitors) to incubate the rat neural stem cells to research the expression of downstream genes in fluoxetine-treated neural stem cells.Methods To isolate,culture the NSCs derived from E15-16 rats and carried out the real-time RT-PCR detection after respectively incubated NSCs with the phosphatidylinositol-3-kinase(PI3-K) inhibitor (LY294002),the mitogen-activated protein kinase kinase(MEK) inhibitor (PD98059) and fluoxetine.Results The fluoxetine can upregulated the mRNA expression of BDNF,GDNF,LIF and BCL-2 in rats NSCs and can not regulated the mRNA expression of CNTF and ILGF1/2.Conclusion The mRNA expression of BDNF,GDNF,LIF and BCL-2 can be up-regulated by fluoxetine through PI3K and(or) MAPK mediated pathway in NSCs.BDNF,GDNF,LIF and BCL-2 not CNTF and ILGF1/2 were the downstream signaling molecule targets of fluoxetine.