目的 研究体外激活的Notch信号系统对许旺细胞的调控作用.方法 培养成年SD大鼠坐骨神经许旺细胞,分别加入Notch信号激活剂Recombinant rat jagged1/FC chimera和抑制剂DAPT,空白对照加PBS缓冲液.通过免疫荧光检测细胞Notch信号蛋白激活状态,MTT检测细胞增殖情况,酶联免疫吸附试验检测细胞分泌NGF情况.结果 倒置显微镜下观察Notch激活组细胞生长优于其他各组;MTT检测该组促细胞增殖作用明显(P<0.05),且增殖效果在72 h内逐渐增强;ELISA显示Notch激活组细胞培养上清液中NGF蛋白浓度[(60.11±2.61)pg/ml]较对照组[(51.29±3.59)pg/ml]及抑制组[(43.43±2.82 pg/ml]明显升高(P<0.05).结论 一定剂量的Notch信号激活剂不仅促进体外培养的许旺细胞增殖,而且促进许旺细胞分泌NGF增多.
Objective To study the effect of activated Notch signaling system on Schwann cells in vitro. Methods Schwann cells were isolated from sciatic nerves of adult SD rats. Recombinant rat jagged1/FC chimera, an activator of the Notch signaling system, and γ-secretase inhibitor (DAPT), an inhibitor of the Notch signaling system, were added into the culture medium respectively. The cells in the medium added with phosphate buffered saline were used as control group. Then the cultured cells were collected. NICD, as a mark of activated Notch system, was detected by immunofluorescence methods. MTT method was used to calculate the growth curve. The level of NGF in the cell supernatant was assayed by using ELISA. Results Schwann cells in recombinant rat jagged/FC chimera group grew better than those in the control groups. MTT method showed that activated Notch signaling significantly promote the proliferation of the cells(P 〈 0.05). After cultured for 48 h, the amount of NGF in cell supernatant of recombinant rat jagged1/FC chimera group was significantly increased (P 〈 0.05). Conclusion When the Notch signaling system is activated, Schwann cells not only proliferate but also secret more NGF.