一氧化氮信号通路对小鼠鼻腔纤毛运动的调控

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

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。

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.