Mono-(2-ethylhexyl) Phthalate Induces Differentiation To Oligodendrocytes In Rat C6Glioma Cells

Objective: Di-(2-ethylhexyl)-phthalate(DEHP) is the most widely used plasticizers indaily-life products.Recently,empirical data have shown that total exposures to DEHP andMEHP are higher in infants and young children than in adults.A experiment shows thatMEHP could inhibit proliferation and promote differentiation in PC12cells,and aepidemiological survey shows that higher total di-2-ethylhexyl phthalate(DEHP)metabolites at26w were associated with more nonoptimal reflexes. In this study we usedthe C6cells line to evaluate the influence of mono-(2-ethylhexyl) phthalate (MEHP), theactive metabolite of DEHP, on the differentiation of glioma cells in vitro.Methods: C6cells were cultured and exposed to MEHP at three consentrations (1mg/L,10mg/L,50mg/L,100mg/L),respectively for24h and48h,along with control group(0.01%DMSO).Then,we determined cell viability by MTT method, and cell apotosis by FACSanalysis.we observed Morphological Transformation of C6Cells,quantified neurite outgr-owth in differentiating C6cells,counted and ealeulated percentage of differentiated cells onan inverted light microscope fitted with phase contrast optics.we determined the expressionof myelin basic protein (MBP),fibrillary acidic protein (GFAP),Peroxisome ProliferatorActivated Receptor γ(PPARγ) by immunofluoresce-nce method and western blot.Resμlts:1. MTT and FACS analysis showed that MEHP had impact on C6cell viability andinhibition cell growth,but there were not dose-and time-dependent effect on cell apotosis. 2.on the inverted light microscope, most of the undifferentiated C6cells in the controlgroup took the shake of long fusiform, some of them are polygonal,and the cell body hasshort and thick processes.compared to the control group,the cell body was more smooth,more round,both ends of the protrusions became finer longer in the MEHP-treated groups;with the increase of MEHP dose and time, the percentage of differentiatedcells(extensions-positive) increase.3.immunofluorescence and western blot analysis showedthat there is no significant differences in the expression of PPARγ and GFAP,but MEHPincrease the expression of MBP.Conclusion: MEHP induces differentiation to oligodendrocytes in rat C6glioma cells.