中华皮肤科杂志 ›› 2004, Vol. 37 ›› Issue (9): 534-536.

• 论著 • 上一篇    下一篇

降钙素基因相关肽对HaCaT细胞一氧化氮合酶表达的影响

聂本勇, 范青源, 郑茂荣, 陶苏江, 顾军, 高春芳, 牟贤龙   

  1. 第二军医大学长海医院皮肤科, 上海200433
  • 收稿日期:2003-09-18 出版日期:2004-09-15 发布日期:2004-09-15
  • 基金资助:
    国家自然科学基金资助项目(3000149)

Effect of CGRP on Expression of Neuronal Nitric Oxide Synthase in HaCaT Cells

NIE Ben-yong, FAN Qing-yuan, ZHENG Mao-rong, TAO Su-jiang, GU Jun, GAO Chun-fang, MU Xian-long   

  1. Department of Dermatology, Changhai Hospital, Second Military Medical University, Shanghai 200433, China
  • Received:2003-09-18 Online:2004-09-15 Published:2004-09-15

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摘要： 目的 研究降钙素基因相关肽对体外培养角质形成细胞HaCaT株神经型一氧化氮合酶(NOS)mRNA、蛋白表达和NO释放的调节作用。方法 应用NO试剂盒(酶法)检测培养细胞上清液中NO水平,RT-PCR和免疫组化(SP)方法检测HaCaT细胞神经型NOSmRNA和蛋白的表达水平。结果 体外正常培养的HaCaT细胞表达和释放基础水平的神经型NOS和NO,降钙素基因相关肽上调HaCaT细胞神经型NOS表达和NO释放,降钙素基因相关肽受体抑制剂CGRP-8-37抑制降钙素基因相关肽的作用。结论 降钙素基因相关肽诱导角质形成细胞表达神经型NOS并促进神经型NO释放。

关键词: 降钙素基因相关肽, 一氧化氮合酶, 一氧化氮

Abstract: Objective To investigate the regulatory effect of calcitonin gene-related peptide (CGRP) on the mRNA and protein expression of neuronal nitric oxide synthase (NOS), as well as the release of nitric oxide (NO) in cultured human keratinocyte line HaCaT. Methods NO level in the supernatant of cell culture medium was detected with an enzymatic NO detecting kit, the mRNA expression of neuronal NOS was studied with reverse transcription polymerase chain reaction(RT-PCR), and the protein expression of neuronal NOS was studied with immunochemical technique(SP). Results Compared with that in normal culture condition, the mRNA and protein expression of neuronal NOS and the release of NO was significantly upregulated by CGRP in HaCaT cells. Whereas, the expression of neuronal NOS and the release of NO in HaCaT cells induced by CGRP was inhibited by CGRP-8-37, an inhibitor of CGRP receptor. Conclusion The expression of neuronal NOS in keratinocytes and the release of NO from keratinocytes could be upregulated by CGRP.

Key words: Calcitonin gene-related peptide, Nitric-oxide synthase, Nitric oxide