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一氧化氮合酶底物L-精氨酸对气管纤毛摆动频率的调节

Regulation of tracheal ciliary beat frequency by nitric oxide synthase substrate L-arginine.

作者信息

Jiao Jian, Han Demin, Meng Na, Jin Shanzhe, Zhang Luo

机构信息

Laboratory of Otolaryngology, Head and Neck Surgery, Ministry of Education, Beijing Institute of Otolaryngology, Beijing, China.

出版信息

ORL J Otorhinolaryngol Relat Spec. 2010;72(1):6-11. doi: 10.1159/000265683. Epub 2010 Jan 22.

Abstract

OBJECTIVES

Our purpose was to investigate the role of nitric oxide (NO) synthase substrate L-arginine (L-arg) in the regulation of airway ciliary beat frequency (CBF).

METHODS

We studied the effects of L-arg on CBF of rabbit tracheal epithelial cells by using high-speed digital microscopy and determined the expression and distribution of neuronal nitric oxide synthase (nNOS), inducible nitric oxide synthase (iNOS) and endothelial nitric oxide synthase (eNOS) in intact tracheal mucosa and in isolated cilia by using immunohistochemistry and Western blot, respectively.

RESULTS

L-arg dose- and time-dependently increased CBF compared with the control groups, and the effect was blocked by pretreatment with the NOS inhibitor N(G)-nitro-L-arginine methyl ester (L-NAME). Immunoreactivity to 3 isoforms of NOS (nNOS, iNOS, and eNOS) was observed in cilia of epithelial cells, and the signal intensity of eNOS was stronger than that of both nNOS and iNOS. Western blot analysis of isolated ciliary preparations also detected all 3 isoforms of NOS, and the expression level is eNOS > nNOS > iNOS.

CONCLUSION

L-arg increases the CBF of rabbit tracheal epithelial cells via the NOS-NO signal pathway.

摘要

目的

我们的目的是研究一氧化氮(NO)合酶底物L-精氨酸(L-arg)在气道纤毛摆动频率(CBF)调节中的作用。

方法

我们通过高速数字显微镜研究了L-arg对兔气管上皮细胞CBF的影响,并分别使用免疫组织化学和蛋白质印迹法测定了完整气管黏膜和分离纤毛中神经元型一氧化氮合酶(nNOS)、诱导型一氧化氮合酶(iNOS)和内皮型一氧化氮合酶(eNOS)的表达和分布。

结果

与对照组相比,L-arg剂量和时间依赖性地增加了CBF,并且该作用被一氧化氮合酶抑制剂N(G)-硝基-L-精氨酸甲酯(L-NAME)预处理所阻断。在上皮细胞的纤毛中观察到对3种一氧化氮合酶同工型(nNOS、iNOS和eNOS)的免疫反应性,并且eNOS的信号强度强于nNOS和iNOS两者。对分离的纤毛制剂进行蛋白质印迹分析也检测到了所有3种一氧化氮合酶同工型,并且表达水平为eNOS>nNOS>iNOS。

结论

L-arg通过一氧化氮合酶-NO信号通路增加兔气管上皮细胞的CBF。

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