KISEP Rhinology Korean J Otolaryngol 2006;49:307-12 사람정상코점막상피세포에서겔형성점소 (Gel-Forming Mucin) 분비에대한 Prostaglandin E 2 의효과 김진국 1 김창훈 2,3 김규보 2 장현종 1 김현직 2 윤주헌 2,3,4 Effect of Prostaglandin E 2 on Gel-forming Mucin Secretion in Normal Human Nasal Epithelial ells Jin Kook Kim, MD 1, hang Hoon Kim, MD 2,3, Kyubo Kim, MD 2, Hyun Jong Jang, MD 1, Hyun Jik Kim, MD 2 and Joo Heon Yoon, MD 2,3,4 1 Department of Otorhinolaryngology-Head and Neck Surgery, KonKuk University ollege of Medicine, Seoul and 2 Department of Otorhinolaryngology 3 The Airway Mucin Institute 4 Brain Korea 21 Project for Medical Sciences, Yonsei University ollege of Medicine, Seoul, Korea ABSTRAT Background and Objectives:Recently, Prostaglandin E 2 ( ) was found to induce MU5A production via an agonist of E-prostanoid (EP2/EP4), but not EP1/EP3, in normal human airway epithelium. However, the receptor that mediates MU5A has not been determined. This study was aimed to investigate the MU5A mucin gene and mucin secretion by and its receptors in cultured normal human nasal epithelial cells. Materials and Method:After treatment with and/or antagonist, gel-forming mucin mrna expression was determined by reverse transcription-polymerase chain reaction. Total mucin and MU5A mucin levels were measured using an immuno-dot blotting assay. Results: increased the MU5A gene expressions and MU5A mucin, but not the expressions of other gel-forming mucin genes. An EP2 receptor antagonist (AH6809) did not suppress the -induced MU5A gene expression or MU5A mucin. However, an EP4 receptor antagonist (AH23848) significantly suppressed the level of -induced MU5A gene expression and MU5A mucin. onclusion:these findings indicate that induces MU5A gene expression and mucin secretion via EP4 receptor in cultured normal human nasal epithelial cells. (Korean J Otolaryngol 2006;49:307-12) KEY WORDS:Prostaglandin E 2 Mucin Secretion Prostaglandin receptor. 307
로유도된점소유전자발현과분비 - - - Table 1. Designed primers used for PR amplification MU5A MU5B MU6 MU7 EP2 EP4 2M Name Sequence Annealing temperature ycle Size F- GA AA TA TT TG GG TG R- GA T AT TT T GT GTT F- AT A GAG AT GT A A R- TA A TGG TT GTG TG TA F- TA T AT A AA AA R- GGA GAA GAA GGA AAA AGA G F- A A TA ATT TT R- TA TTG T A AT GT F- GA GA GG AA GTG GT T R- GA GA GG GG TG GTA GA F-TG G AAG GAG AG AAG GAG A R-GA GGT GG GAG AAT GAG GAA GGA F- TG G TA TT T TTT TG G R- GT TA ATG TT GA T A TTA A 60 33 360 bp 55 30 338 bp 55 30 293 bp 55 30 209 bp 60 30 409 bp 60 30 468 bp 55 28 360 bp 308 Korean J Otolaryngol 2006;49:307-12
김진국외 - - t EP2 EP4 2M 2 7 14 21 Days after confluence Fig. 1. EP2 gene expression increased as a function of mucociliary differentiation. However, EP4 gene expression was not altered as a function of differentiation. 110-7 M AH 6809 110-5 M AH 23848 110-5 M MU5A 1.8 1.6 MU5B MU6 MU7 2M Relative intensities 1.4 1.2 0.1 0.8 0.6 0.4 0.0 AH6809 AH23848 Fig. 2. Effects of prostaglandin E 2 on gel-forming mucin gene expressions and inhibitory effects of receptor antagonists. increased only MU5A gene expression, but not the expression other gelforming mucin genes. An EP2 receptor antagonist AH6809 did not suppress -induced MU5A gene expression. However, an EP4 receptor AH23848 antagonist significantly suppressed MU5A gene expression. 309
로유도된점소유전자발현과분비 Fold increase of total mucin 0.3 2.5 1.5 0.1 0.5 0.0 p0.05 p0.01 AH6809 AH23848 Fold increase of MU5A mucin 0.5 4.5 0.4 3.5 0.3 2.5 1.5 0.1 0.5 0.0 p0.05 p0.01 AH6809 AH23848 Fig. 3. Effect of on total mucin and MU5A mucinsecretion. increased total mucin secretion and promoted a 4-fold increase of MU5A mucin secretion. An EP4 antagonist AH23848 significantly suppressed both total mucin and MU5A mucin secretion. EP2 EP4 2M Fig. 4. Detection of EP E prostanoid mrna expression in cultured NHNE cells. did not affect EP2 or EP4 gene expression. 310 Korean J Otolaryngol 2006;49:307-12
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