一氧化氮信号通路对小鼠鼻腔纤毛运动的调控

目的观察一氧化氮(nitric oxide,NO)信号系统对小鼠鼻腔纤毛运动的调控。方法 酶消化法培养小鼠鼻腔上皮细胞,采用高速数字化显微成像技术对纤毛摆动进行定量分析,观察10mmol/LL-精氨酸(L-arginine,L-arg)对上皮细胞纤毛摆动的影响;采用免疫组化方法检测NO信号系统分子内皮型一氧化氮合酶(endothelial nitric oxide synthase,eNOS)、鸟苷酸环化酶(guanylate cyclase,GC)、蛋白激酶G(protein kinase G,PKG)的表达。结果 ①Hanks平衡盐溶液(Hank’s balanced salt solution,HBSS)对照组小鼠鼻纤毛摆动在10min内无明显变化;②10mmol/L L-arg处理组纤毛摆动频率(ciliary beat frequency,CBF)加药后2min即出现明显增加,在检测的各个时间点,CBF与加药前相比差异有统计学意义;③免疫组化结果显示NO信号通路中主要信号分子eNOS、GC、PKG在小鼠鼻腔纤毛中均有明显表达。结论 NO-环磷酸鸟苷(cyclic guanosine monophosphate,cGMP)-PKG信号通路参与了小鼠鼻纤毛运动的调控。

Regulatory Effect of Nitric Oxide Signal Pathway on Ciliary Motility in Mouse Nasal Epithelial Cells

Objective To investigate the effects of nitric oxide(NO) signal pathway on ciliary beat frequency(CBF) in mouse nasal epithelial cells.
Methods Cultured mouse nasal epithelial cells were treated with 10 mmol/L L-arginine, the CBF was measured by high-speed digital microscopy. In addition, expression of endothelial nitric oxide synthase(eNOS), guanylate cyclase(GC) and protein kinase G(PKG) were detected by immunohistochemistry.
Results ① CBF of epithelial cells in Hank’s balanced salt solution(HBSS) control group showed no significant changes in 10 min. ② CBF increased with treatment of 10 mmol/L L-arginine at 2 min and significant changes were observed at all the time points detected. ③ Immunoreactivity to eNOS, GC and PKG were observed in cilia of epithelial cells.
Conclusion NO-cGMP-PKG signal pathway participates in the regulating mechanism of ciliary motility in mouse nasal epithelial cells.