关键词: 地黄多糖；骨髓间充质干细胞；Notch信号通路；诱导；神经样细胞

Abstract:

Objective   To explore the effect of rehmannia glutinosa olysaccharide (RGP) on rat bone marrow mesenchymal stem cells (BMSCs) differentiation into neuron-like cells and Notch signaling pathway. Methods   Rat BMSCs were experimented as the object, the P3-P5 generations of BMSCs were randomly divided into the control group and RGP group(0.2mg/mL), and induced and cultured for 7 days. Immunocytochemistry assay and Western blotting method were used to detect Notch1, Jagged1 protein and intracellular domain of Notch 1(NICD) expressions at 4 point in time (0,1,3 and 7day) after the induction. The molecular mRNA expression and changes related to the Notch signaling pathway were analyzed by Real-time PCR. Results   Immunofluorescence assay showed that the Notch1 protein expression of BMSCs decreased over time in the RGP group, positive cell percentage decreased likewise, which was significantly different from the control group(P＜0.01). The Jagged1 positive cell rate of RGP group increased from induced end 0d to 1d after the induction, decreased significantly from 1d to 7d, and there was significant difference between each time point comparison(P＜0.05), also different from the control group(P＜0.05). Western blotting results were similar to that of the immunofluorescence method. Real-time PCR showed that Notch1 mRNA expression decreased over time. Presenilin1 decreased and then went up mildly afterwards, Hes1 decreased, Mash1 increased, and Jagged1 increased then decreased in the RGP group, which were significantly different from the control group at all durations(P＜0.01). Conclusion    RGP might restrain Notch1 protein expression during BMSCs differentiations into neuron-like cells in vitro, and influence the related molecular mRNA expression of the Notch signaling pathway.

Key words: Rehmannia glutinosa polysaccharide; Bone marrow mesenchymal stem cells; Notch signal pathway; Induction; Nerve-like cells