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Expression of Na+/H+ exchanger isoforms in normal human nasal epithelial cells and functional activity of Na+/H+ exchanger 1 in intracellular pH regulation

 Ji-Hyun Shin  ;  Wan Namkung  ;  Joo-Heon Yoon  ;  Min Goo Lee  ;  Jeung-Gweon Lee  ;  Kyung-Dong Lee  ;  Jong-Bum Yoo  ;  Jae Young Choi  ;  Chang-Hoon Kim 
 ACTA OTO-LARYNGOLOGICA, Vol.125(3) : 286-292, 2005 
Journal Title
Issue Date
Blotting, Western ; Cation Transport Proteins/antagonists & inhibitors ; Cation Transport Proteins/metabolism* ; Cell Differentiation ; Cells, Cultured ; Epithelial Cells/metabolism* ; Epithelial Cells/pathology ; Guanidines/pharmacology ; Humans ; Hydrogen-Ion Concentration ; Intracellular Fluid/chemistry* ; Membrane Proteins/antagonists & inhibitors ; Membrane Proteins/metabolism* ; Nasal Mucosa/cytology* ; Protein Isoforms/metabolism ; RNA, Messenger/metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Sodium-Hydrogen Exchanger 1 ; Sodium-Hydrogen Exchangers/antagonists & inhibitors ; Sodium-Hydrogen Exchangers/metabolism* ; Sulfones/pharmacology
CONCLUSIONS: Both the mRNA and protein of NHE1, -2 and -3 were expressed in NHNE cells. NHE1 is a major NHE isoform which is expressed in the basolateral membranes of NHNE cells.
OBJECTIVE: Na+/H+ exchangers are ubiquitous plasma membrane transport proteins implicated in the maintenance of intracellular pH. The aim of this study was to examine the expression of Na+/H+ exchangers as a function of the differentiation of normal human nasal epithelial (NHNE) cells. In addition, we investigated the functional activity of the Na+/H+ exchanger 1 (NHE1) in basolateral membranes.
MATERIAL AND METHODS: Cultured passage-2 NHNE cells were used. RNA and histological samples were collected on the day of confluence and on the 7th, 14th and 28th days after confluence in order to determine the effects of time. Cell lysates were collected on the day of confluence to investigate the presence of the proteins. Reverse transcriptase polymerase chain reaction and Western blotting were performed to investigate the presence of mRNA and protein, respectively. The functional activity of NHE1 was examined using 3-methylsulfonyl-4-piperidinobenzoyl guanidine methanesulfonate (HOE694), an NHE1-specific inhibitor, on the day of confluence.
RESULTS: The NHE1 mRNA expression level did not change as a function of differentiation. However, the NHE2 mRNA expression levels increased on the 7th, 14th and 28th days after confluence. The NHE3 mRNA expression levels increased on the 14th and 28th days after confluence. Western blot analysis confirmed the expression of NHE1 (91 kDa), NHE2 (90 kDa) and NHE3 (93 kDa). In addition, HOE694 inhibited basolateral NHE activity by 68% at 1 microM and by 85% at 5 microM in the NHNE cells.
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1. College of Medicine (의과대학) > Dept. of Otorhinolaryngology (이비인후과학교실) > 1. Journal Papers
1. College of Medicine (의과대학) > Dept. of Pharmacology (약리학교실) > 1. Journal Papers
1. College of Medicine (의과대학) > Yonsei Biomedical Research Center (연세의생명연구원) > 1. Journal Papers
Yonsei Authors
Kim, Chang Hoon(김창훈) ORCID logo https://orcid.org/0000-0003-1238-6396
Shin, Ji Hyun(신지현)
Yoo, Jong Bum(유종범)
Yoon, Joo Heon(윤주헌)
Lee, Min Goo(이민구) ORCID logo https://orcid.org/0000-0001-7436-012X
Lee, Jeung Gweon(이정권)
Choi, Jae Young(최재영) ORCID logo https://orcid.org/0000-0001-9493-3458
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