The identification of the TRPM8 channel on primary culture of human nasal epithelial cells and its response to cooling

Shao Cheng Liu, Hsuan Hsuan Lu, Hueng Chuen Fan, Hsing Won Wang, Hang Kang Chen, Fei Peng Lee, Chong Jen Yu, Yueng Hsiang Chu

Research output: Contribution to journalArticle

Abstract

Background: It has been proposed that the transient receptor potential (TRP) channel Melastatin 8 (TRPM8) is a cold-sensing TRP channel. However, its presence and its role in the nasal cavity have not yet been fully studied. Methods: Immunohistology was used to study TRPM8 receptors in both the nasal mucosa tissue and the primary cultures of human nasal cells. Cells from primary cultures were immunostained with antibodies to TRPM8, mucin, cytokeratin (CK)-14, CK-18, and vimentin. Western blotting and real-time polymerase chain reaction (PCR) were used to determine the physiological role of TRPM8 in mucus production in the nasal cavity, with and without its agonist and antagonist. Results: The TRPM8 is clearly present in the epithelium, mucous glands, and vessels. No obvious TRPM8-immunoreactive cells were detected in the connective tissue. Immunostaining of cytospin preparations showed that epithelial cells test positive for CK-14, CK-18, TRPM8, and mucin 5AC (MUC5AC). Fibroblastic cells are stained negative for TRPM8. Secreted mucins in the cultured supernatant are detected after exposure to menthol and moderate cooling to 24°C. Both induce a statistically significant increase in the level of MUC5AC mRNA and mucin production. BCTC, a TRPM8 antagonist, has a statistically significant inhibitory effect on MUC5AC mRNA expression and MUC5AC protein production that is induced by menthol and moderate cooling to 24°C. Conclusions: The study demonstrates that TRPM8 is present in the nasal epithelium. When it is activated by moderate cooling to 24°C or menthol, TRPM8 induces the secretion of mucin. This study shows that TRPM8 channels are important regulators of mucin production. Therefore, TRPM8 antagonists could be used to treat refractory rhinitis.

Original languageEnglish
Article numbere7640
JournalMedicine (United States)
Volume96
Issue number31
DOIs
Publication statusPublished - Aug 1 2017

Fingerprint

Mucin 5AC
Mucins
Nose
Menthol
Epithelial Cells
Keratin-14
Keratin-18
Transient Receptor Potential Channels
Nasal Mucosa
Nasal Cavity
Messenger RNA
Primary Cell Culture
Vimentin
Mucus
Rhinitis
Connective Tissue
Real-Time Polymerase Chain Reaction
Epithelium
Western Blotting
Antibodies

Keywords

  • cold receptor
  • immunohistochemistry
  • menthol
  • mucin
  • TRPM8

ASJC Scopus subject areas

  • Medicine(all)

Cite this

The identification of the TRPM8 channel on primary culture of human nasal epithelial cells and its response to cooling. / Liu, Shao Cheng; Lu, Hsuan Hsuan; Fan, Hueng Chuen; Wang, Hsing Won; Chen, Hang Kang; Lee, Fei Peng; Yu, Chong Jen; Chu, Yueng Hsiang.

In: Medicine (United States), Vol. 96, No. 31, e7640, 01.08.2017.

Research output: Contribution to journalArticle

Liu, Shao Cheng ; Lu, Hsuan Hsuan ; Fan, Hueng Chuen ; Wang, Hsing Won ; Chen, Hang Kang ; Lee, Fei Peng ; Yu, Chong Jen ; Chu, Yueng Hsiang. / The identification of the TRPM8 channel on primary culture of human nasal epithelial cells and its response to cooling. In: Medicine (United States). 2017 ; Vol. 96, No. 31.
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AU - Liu, Shao Cheng

AU - Lu, Hsuan Hsuan

AU - Fan, Hueng Chuen

AU - Wang, Hsing Won

AU - Chen, Hang Kang

AU - Lee, Fei Peng

AU - Yu, Chong Jen

AU - Chu, Yueng Hsiang

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AB - Background: It has been proposed that the transient receptor potential (TRP) channel Melastatin 8 (TRPM8) is a cold-sensing TRP channel. However, its presence and its role in the nasal cavity have not yet been fully studied. Methods: Immunohistology was used to study TRPM8 receptors in both the nasal mucosa tissue and the primary cultures of human nasal cells. Cells from primary cultures were immunostained with antibodies to TRPM8, mucin, cytokeratin (CK)-14, CK-18, and vimentin. Western blotting and real-time polymerase chain reaction (PCR) were used to determine the physiological role of TRPM8 in mucus production in the nasal cavity, with and without its agonist and antagonist. Results: The TRPM8 is clearly present in the epithelium, mucous glands, and vessels. No obvious TRPM8-immunoreactive cells were detected in the connective tissue. Immunostaining of cytospin preparations showed that epithelial cells test positive for CK-14, CK-18, TRPM8, and mucin 5AC (MUC5AC). Fibroblastic cells are stained negative for TRPM8. Secreted mucins in the cultured supernatant are detected after exposure to menthol and moderate cooling to 24°C. Both induce a statistically significant increase in the level of MUC5AC mRNA and mucin production. BCTC, a TRPM8 antagonist, has a statistically significant inhibitory effect on MUC5AC mRNA expression and MUC5AC protein production that is induced by menthol and moderate cooling to 24°C. Conclusions: The study demonstrates that TRPM8 is present in the nasal epithelium. When it is activated by moderate cooling to 24°C or menthol, TRPM8 induces the secretion of mucin. This study shows that TRPM8 channels are important regulators of mucin production. Therefore, TRPM8 antagonists could be used to treat refractory rhinitis.

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KW - immunohistochemistry

KW - menthol

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KW - TRPM8

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