目的:观察去甲肾上腺素(norepinephrine,NE)对骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)增殖的影响及其作用途径。方法:分离培养正常大鼠BMSCs,采用3H-TdR掺入实验检测不同浓度的NE(10-7-10-4M)作用8h及10-5M的NE作用不同时间(0-24h)BMSCs细胞增殖情况,realtimeRT-PCR检测肾上腺素能受体α1A-AR,α1B-AR和α1D-ARmRNA表达变化情况。结果:10-7-10-4M的NE作用8h后均促进了BMSCs细胞的增殖,并且在10-5M时NE对BMSCs的促增殖效应最为显著;正常组BMSCs细胞的α1A-AR,α1B-AR,α1D-ARmRNA表达维持在较低水平,加入10-5M的NE作用后α1-AR三个亚型mRNA表达水平均有不同程度的升高(P〈0.05)。结论:NE能够促进BMSCs的增殖,并且这种促增殖作用是通过AR依赖的信号通路来调节的。
Objective: To observe the effects of norepinephrine on bone marrow mesenchymal stem cells proliferation and the re- lated signaling molecules involved in BMSCs. Methods: To investigate whether norepinephrine induced BMSCs proliferation, [3H] thymidine incorporation into BMSCs was tested in the presence of various concentrations of norepinephrine (10-7-10-4 M) for 8 h or a fixed concentration of norepinephrine (10-5 M) at various time points (0-24h). The levels of α1A-AR,α1B-AR and α1D-AR mRNA ex- pression were assessed by real-time RT-PCR. Results: Norepinephrine treatment (10-7-10-4 M) for 8 h increased [3H]thymidine incorpo- ration in a dose-dependent manner. The maximal stimulatory effect was observed at 10-5 M of norepinephrine. BMSCs maintained in basal medium had low levels of α1A-AR, α1B-AR and α1D-AR mRNA expression. Norepinephrine upregulated mRNA expression of different AR subtypes in BMSCs (P 〈 0.05). Conclusion: This stimulatory effect of norepinephrine on cell proliferation was mediated through AR-dependent signaling.