The Preliminary Study On The Genesis Of Jagged1 MRNA In Astrocyte Induced By TGF-β1

Objective:This study was designed to investigate on the genesis of Jagged1 mRNA in astrocyte induced by TGF-β1 and discusses possible mechanism for multiple sclerosis (MS) which may be as a new theoretical basis for MS.Methods:1. Astrocyte cell culture: With the 1 ~ 2 d C57BL/6 newborn mice, brainstems were stripped, digested, and by some other steps,the early cells were cultivated to poly-lysine-coated culture flasks, the purified astrocytes were collected by subculture. And use specific antibodies to detect GFAP+ astrocytes.2. TGF-β1 stimulation: Astrocytes activated by LPS were cultivated with TGF-β1 during 24 hours, then extracted the Jagged1 mRNA. The expression of Jagged1 mRNA was evaluated at 0 h,6 h,12 h and 24 h.Results:1. GFAP immunofluorescence staining showed that the GFAP positive cell were in majority.2. TGF-β1 induced the expression of Jagged1 mRNA in astrocytes: With stimulus duration extending, the expression of Jagged1 mRNA attenuated significantly.Conclusion:1. Successfully cultured primary astrocytes (Figure 1) and the purity was over 95%.2. The expression of Jagged1 mRNA in astrocytes that activated by LPS, which induced by TGF-β1, maybe lead to activation of Jagged1/Notch pathway and inhibit the repair of myelin sheath.