Retinoic acid enhances expression of neural specific genes in Sca-1^+ cells of mouse fetal liver through activating protein kinase C

摘要

BACKGROUND: Interstitial stem cell is characterized by multiple differentiations, and retinoic acid (RA) can induce differentiation of stromal cells into nerve tissue cells in fetal liver of mice, so, its signal transduction pathway should be discussed to trigger differentiation. OBJECTIVE: To study the effect of RA on expression of neural specific gene and its signal transduction in fetal liver of mice. DESIGN: Paired controlled study on the basis of cell. SETTING: Institute of Hematology, Medical College of Jinan University. MATERIALS: The experiment was completed in the Institute of Hematology, Medical College of Jinan University from April to December 2005. C57BL/6 mice, of clean grade, aged 8-10 weeks, weighting 20-35 g, 10 females and 4 males, were selected in this study. METHODS: Sca-1+ cells in fetal liver were prepared with MACS kit and cultured with DMEM + 10% fetal bovine serum (FBS). On the fourth day, it was added with or without protein kinase C (PKC) inhibitor chelerythrine chloride (3 μmol/L) and 5×10-7 mol/L RA for 24 hours, and then incubated in serum-free medium for 5 days. Expressions of genes were assayed by Western blotting and semi-quantitative reverse transcription polymerase chain reaction. MAIN OUTCOME MEASURES: Expression of neural specific gene NF-L, NF-H, BF-1 and TH. RESULTS: Expression of neural specific gene NF-L, NF-H, BF-1 and TH was significantly increased after treatment with RA and they were increased 5.06, 5.15, 4.63 and 3.33 times, respectively. However, chelerythrine chloride could inhibit expression of neural specific gene NF-L, NF-H, BF-1 and TH induced by RA. CONCLUSION: RA can promote the expression of neural specific genes in Sca-1+ cells of fetal liver, and its pathway may be related to PKC.