The Influence Of Propentofylline On Cytokines Secretion Of Cerebral Cortical Astrocyte In Vitro

Objective To investigate the effect of PPF(propentofylline,PPF)on the cytokines(NGF,IL-1β)secretion and GFAP(glial fibrillary acidic protein,GFAP) expression of cerebral cortical astrocytes from newborn SD rat,and to learn whether LPS(lipopolysaccharide,LPS)could affect the influence of propentofylline on the secretion of astrocytes.Methods Primarily cultured astrocytes with a purity of more than 95%from neonatal SD rat cerebral cortical tissue were used.These astrocytes were plated into cell culture plates at a density of 1×10~5 cells/ml after passage three times.The cells were divided into 8 groups(n=6 in each group):control group,PPF10μmol/L group,PPF100μmol/L group,PPF1000μmol/L group,LPS1μg/mL group,PPF10μmol/ L+LPS1μg /mL group,PPF100μmol/L+LPS1μg/mL group and PPF1000μmol/L+ LPS 1μg/mL group.The cells were treated with PPF and/or LPS for 1 and 3 days.Sandwich ELISA was selected for quantitative determination of IL-1βand NGF protein in cell culture supernatant.Software Image-Pro Plus(Image-Pro Plus,IPP)was used for the measurement of IOD(integrated optical density,IOD)and AREA of GFAP positive region from immunohistochemistry stain.Results Compare with the results in the control group,PPF increase the secretion of NGF by AST(P＜0.01),LPS decrease the secretion of NGF(P＜0.05),PPF increase the secretion of NGF by AST which was activated by LPS(P＜0.01);Compare with the results in the control group,PPF decrease the secretion ofIL-1βby AST(P＜0.01),LPS increase the secretion of IL-1β(P＜0.01),PPF of high dosage groups(100,1000μmol/L)can decrease the secretion of IL-1βby AST which was activated by LPS(P＜0.01),but cannot the low dosage(10μmol/L)group.IOD and AREA of GFAP positive region could be diminished by PPF significantly compared with those of control group.LPS could increase IOD and AREA significantly compared with those of control group.Conclusion The results suggested that LPS may activate astrocytes,increase the secretion of IL-1βand decrease the secretion of NGF from astrocytes,enlarge IOD and AREA of GFAP positive region.Propentofytline might decrease the secretion of IL-1βand increase the secretion of NGF,diminish IOD and AREA.Propentofylline could lighten the activation of LPS to astrocytes.