羧甲基壳聚糖对体外培养许旺细胞环磷腺苷/蛋白激酶A信号通路的影响

Carboxymethylated chitosan effect on cyclic adenosine monophosphate/protein kinase A signaling pathway in rat Schwann cells cultured in vitro

背景:已有研究证实羧甲基壳聚糖对许旺细胞增殖、分泌具有促进作用,但其对许旺细胞内环磷腺苷介导蛋白激酶A信号通路的影响仍需要进一步研究。目的:观察羧甲基壳聚糖对大鼠许旺细胞内环磷腺苷/蛋白激酶A信号通路的影响。方法:取第2代新生大鼠许旺细胞,以1×109 L-1的细胞浓度接种于6孔板,分4组培养,空白对照组加入PBS,实验组分别加入50,100,200 mg/L的羧甲基壳聚糖溶液培养。培养24 h后,检测许旺细胞内环磷腺苷浓度、蛋白激酶A活性及环磷腺苷反应元件结合蛋白m RNA的表达。结果与结论:与空白对照组比较,羧甲基壳聚糖呈剂量依赖性提高许旺细胞内环磷腺苷浓度(P<0.05),呈剂量依赖性增强许旺细胞内蛋白激酶A活性(P<0.05),呈剂量依赖性提高许旺细胞内环磷腺苷反应元件结合蛋白m RNA的表达(P<0.05)。表明羧甲基壳聚糖可增加许旺细胞内环磷腺苷浓度、促进蛋白激酶A活性,从而激活环磷腺苷/蛋白激酶A信号通路。

BACKGROUND： It has been confirmed that carboxymethylated chitosan has an promoting effect on Schwann cell proliferation and secretion,but its impact on the cyclic adenosine monophosphate-mediated protein kinase A signaling pathway in schwann cell still needs further study. OBJECTIVE： To investigate the effect of carboxymethylated chitosan on cyclic adenosine monophosphate/ protein kinase A signaling pathway in rat schwann cells. METHODS： The Schwann cells of the second generation neonatal rats were obtained and seeded in 6-well plate at a concentration of 1×109/L. These Schwann cells were cultured and divided into four groups. The Schwann cells in the control group were cultured by adding PBS. The Schwann cells in the experimental groups were cultured by adding 50,100 and 200 mg/L of carboxymethyl chitosan solution,respectively. After 24 hours,the concentration of cyclic adenosine monophosphate,protein kinase A activity and cyclic adenosine monophosphate response element binding protein m RNA expression were detected. RESULTS AND CONCLUSION： Compared with the control group,carboxymethyl chitosan increased cyclic adenosine monophosphate concentrations,the activity of protein kinase A and cyclic adenosine monophosphate response element binding protein m RNA expression within the Schwann cells in a dose-dependent manner（P〈0.05）. These results demonstrate that carboxymethyl chitosan can increase the concentration of cyclic adenosine monophosphate within the Schwann cells and promote protein kinase A activity,thereby activating cyclic adenosine monophosphate/protein kinase A signaling pathway.