中国神经再生研究(英文版) ›› 2020, Vol. 15 ›› Issue (2): 352-360.doi: 10.4103/1673-5374.265558

• 原著:颅神经损伤修复保护与再生 • 上一篇    下一篇

内皮素促进大鼠嗅黏膜细胞的增殖

  

  • 出版日期:2020-02-15 发布日期:2020-05-25
  • 基金资助:

    法国国家农业研究所(INRA)资助

Endothelin increases the proliferation of rat olfactory mucosa cells

Bertrand Bryche1, 2, Audrey Saint-Albin1, Claire Le Poupon Schlegel1, Christine Baly1, Patrice Congar1, Nicolas Meunier1, 2   

  1. 1 Neurobiologie de l’olfaction, Institut National de Recherche Agronomique (INRA), Université Paris-Saclay, Jouy-en-Josas, France
    2 Université de Versailles Saint-Quentin en Yvelines, Département de Biologie, Versailles, France
  • Online:2020-02-15 Published:2020-05-25
  • Contact: Nicolas Meunier, PhD,nicolas.meunier@inra.fr.
  • Supported by:
    This work was funded by the Institut National de la Recherche Agronomique (INRA).

摘要:

嗅黏膜为了保持完整性,是动物在整个生命中不断更新的少数神经区之一;在调节这种更新的因子中,内皮素在大鼠嗅上皮中起抗凋亡因子的作用。为探讨内皮素是否也可以作为增殖因子,实验将嗅黏膜进行原代培养,发现内皮素的早期干预促进其生长;而用内皮素受体拮抗剂BQ123和BQ788的混合物干预则抑制了原代培养物的生长,但不影响细胞死亡水平。然后实验使用钙成像,RT-qPCR和蛋白质水平检测显示,内皮素由原代培养物局部合成。此外,大鼠鼻内滴注内皮素受体拮抗剂导致表达细胞增殖标志物增殖细胞核抗原的嗅粘膜细胞减少,且只有短期(1d)内皮素受体拮抗剂鼻内滴注才能降低嗅粘膜细胞的增殖细胞核抗原水平。总之,实验结果表明内皮素有促进嗅黏膜细胞增殖作用,并且该作用是动态调节的。实验于2012年11月28日获得地方伦理委员会(CNREEA#45;协议批准#12-058)的批准。

orcid: 0000-0002-2769-647X (Nicolas Meunier)

关键词:

嗅上皮, 嗅觉, 细胞培养, 自分泌因子, 细胞动力学, 内皮素, 嗅粘膜原代培养, 嗅觉基底细胞

Abstract: The olfactory mucosa holds olfactory sensory neurons directly in contact with an aggressive environment. In order to maintain its integrity, it is one of the few neural zones which are continuously renewed during the whole animal life. Among several factors regulating this renewal, endothelin acts as an anti-apoptotic factor in the rat olfactory epithelium. In the present study, we explored whether endothelin could also act as a proliferative factor. Using primary culture of the olfactory mucosa, we found that an early treatment with endothelin increased its growth. Consistently, a treatment with a mixture of BQ123 and BQ788 (endothelin receptor antagonists) decreased the primary culture growth without affecting the cellular death level. We then used combined approaches of calcium imaging, reverse transcriptase-quantitative polymerase chain reaction and protein level measurements to show that endothelin was locally synthetized by the primary culture until it reached confluency. Furthermore, in vivo intranasal instillation of endothelin receptor antagonists led to a decrease of olfactory mucosa cell expressing proliferating cell nuclear antigen (PCNA), a marker of proliferation. Only short-term treatment reduced the PCNA level in the olfactory mucosa cells. When the treatment was prolonged, the PCNA level was not statistically affected but the expression level of endothelin was increased. Overall, our results show that endothelin plays a proliferative role in the olfactory mucosa and that its level is dynamically regulated. This study was approved by the Comité d’éthique en expérimentation animale COMETHEA (COMETHEA C2EA -45; protocol approval #12-058) on November 28, 2012.

Key words: autocrine factor, cell culture, cellular dynamics, endothelin, olfaction, olfactory basal cells, olfactory epithelium, olfactory mucosa primary culture