目的观察去甲肾上腺素对骨髓间充质干细胞增殖的影响及其作用途径。方法将BMSCs细胞随机分为3组:①正常对照组;②去甲肾上腺素(10-6~10-4mol/L)处理组;③酚妥拉明(10-6mol/L)采+去甲肾上腺素(10-5mol/L)处理组。采用3H-TdR掺入实验检测不同实验组对BMSCs增殖的影响;免疫组化和免疫印记法检测去甲肾上腺素对蛋白激酶C的作用。结果①10-6~10-4mol/L的去甲肾上腺素作用8h后均促进了骨髓间充质干细胞的增殖,并且在10-5mol/L时去甲肾上腺素对骨髓间充质干细胞的促增殖作用最为显著;②酚妥拉明阻断了去甲肾上腺素对骨髓间充质干细胞的促增殖作用;③去甲肾上腺素诱导蛋白激酶C从细胞质到细胞膜转位,酚妥拉明抑制了去甲肾上腺素诱导的蛋白激酶C激活。结论去甲肾上腺素能够促进骨髓间充质干细胞的增殖,并且这种促增殖作用是通过肾上腺素能受体、蛋白激酶C信号通路来调节的。
Objective To investigate the effects of norepinephrine on the proliferation of bone marrow mesenchymal stem cells (BMSCs) and the related signaling molecules involved in BMSCs. Methods 3 H- thymidine incorporation assay was performed to investigate the effect of norepinephrine on BMSCs proliferation, in the presence of various concentrations ( 10^-6 to 10^-4 tool/L) for 8 h or with phentolamine ( 10^-6 tool/L) for 30 min. Effect of norepinephrine on PKC activation was detected by immunofluorescent staining and Western blot analysis. Results Norepinephrine treatment ( 10^-6 to10^-4 tooL/L) for 8 h increased the 3H-thymidine incorporation in a dose-dependent manner. The maximal stimulatory effect was observed at 10^ -5 mol/L of norepinephrine. The stimulatory effect of norepinephrine on cell proliferation was blocked by co-incubation with phentolamine. Norepinephrine induced PKC translocation to the cell membrane, which was blocked by pretreatment with phentolamine for 30 min. Conclusion This stimulatory effect of norepinephrine on cell proliferation is mediated through AR/PKC-dependent signaling.