The Preliminary Study on the Genesis of Jagged1 mRNA in Astrocyte Induced by TGF-β1
|School||Nanjing Medical University|
|Keywords||Astrocytes Jagged1 Transforming growth factor - beta1|
Objective: In this study, astrocytes cultured in vitro model of transforming growth factor - beta1 ( TGF - beta1 ) Jagged1 gene astrocytes , and to explore its possible pathogenesis of multiple sclerosis ( MS ) provide a new theoretical basis for the MS demyelination occurs . Methods: 1. Astrocytes in vitro culture system : the newborn 1 to 2 d C57BL / 6 mice neonatal rat cerebral cortex sterile , remove the meninges , digestive , over the screen and other steps , made ??the initial cell the suspension inoculated flasks polylysine , subcultured to obtain purified astrocytes . Specific antibodies to detect astrocytes and glial fibrillary acidic protein (GFAP) . (2) the establishment of TGF-beta1 -induced astrocyte express Jagged1 system : thoroughly lipopolysaccharide ( LPS ) - activated astrocytes , astrocytes from the resting state into the activated state , and then give the TGF-beta1 cultured for 24 h and extracted astrocytes mRNA , observed at each time point ( 0, 6, 12 and 24 h ) Jagged1 mRNA level . Results: 1 . Cytoskeleton of astrocytes cultured astrocytes ( GFAP ) immunofluorescence staining , showing the vast majority of cells positive for GFAP . 2.TGF - beta1 promote astrocytes Jagged1 mRNA Jagged1 mRNA levels gradually decay astrocytes ( P lt ; 0.05 ) and with prolonged incubation time . Conclusion : 1. Successful primary cultured astrocytes (Fig. 1) . After passage pure culture more than 95% purity . 2.TGF - beta1 to make the activated astrocytes Jagged1 mRNA may cause abnormal Jagged1/Notch pathway activation , which led to the myelin repair disorders .