TY - JOUR
T1 - Effect of doxycycline on transforming growth factor-beta-1'induced matrix metalloproteinase 2 expression, migration, and collagen contraction in nasal polyp'derived fibroblasts
AU - Shin, Jae Min
AU - Park, Joo Hoo
AU - Kang, Byungjin
AU - Lee, Seoung Ae
AU - Park, Il Ho
AU - Lee, Heung Man
N1 - Funding Information:
Funded by a grant from the Korean Health Technology R&D Project, Ministry of Health and Welfare, Republic of Korea (HI15C1512), and by the Basic Science Research Program through the National Research Foundation of Korea, which is funded by the Ministry of Education (2013R1A1A2059716)
Publisher Copyright:
© Oceanside Publications, Inc. All rights reserved.
PY - 2016/11/1
Y1 - 2016/11/1
N2 - Purpose: It is well known that doxycycline has antibacterial and anti-inflammatory effects. In this study, we aimed to investigate the effects of doxycycline on the transforming growth factor (TGF) beta 1'induced matrix metalloproteinase (MMP) 2 expression, migration, and collagen contraction, and to determine its molecular mechanism on nasal polyp' derived fibroblasts (NPDF). Methods: NPDFs were isolated from the nasal polyps of six patients. Doxycycline was used to pretreat TGF-beta-1'induced NPDFs and ex vivo organ cultures of nasal polyps. Cytotoxicity was evaluated by using a 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyl-tetrazolium bromide assay. Smad2/3 is one of the major transcription factors of TGF-beta signaling. The expression levels of MMP2 and Smad2/3 were measured by using Western blotting, reverse transcription'polymerase chain reaction, and immunofluorescence staining. The enzymic activity of MMP2 was analyzed by using gelatin zymography. Fibroblast migration was evaluated by using transwell migration assays. Contractile activity was measured by a collagen gel contraction assay. Results: The expression level of MMP2 in nasal polyp tissues increased in comparison with inferior turbinate tissues. TGF-beta-1'induced NPDFs were not affected by doxycycline (0'40 μg/mL). The expression levels of MMP2 and activation of Smad2/3 in TGF-beta-1'induced NPDFs and in organ cultures of nasal polyps were significantly downregulated with doxycycline pretreatment. Doxycycline also reduced TGF-beta-1'induced fibroblast migration and collagen contraction in NPDFs. Conclusion: Doxycycline inhibited TGF-beta-1'induced MMP2 expression, migration, and collagen contraction via the Smad2/3 signal pathways in NPDFs.
AB - Purpose: It is well known that doxycycline has antibacterial and anti-inflammatory effects. In this study, we aimed to investigate the effects of doxycycline on the transforming growth factor (TGF) beta 1'induced matrix metalloproteinase (MMP) 2 expression, migration, and collagen contraction, and to determine its molecular mechanism on nasal polyp' derived fibroblasts (NPDF). Methods: NPDFs were isolated from the nasal polyps of six patients. Doxycycline was used to pretreat TGF-beta-1'induced NPDFs and ex vivo organ cultures of nasal polyps. Cytotoxicity was evaluated by using a 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyl-tetrazolium bromide assay. Smad2/3 is one of the major transcription factors of TGF-beta signaling. The expression levels of MMP2 and Smad2/3 were measured by using Western blotting, reverse transcription'polymerase chain reaction, and immunofluorescence staining. The enzymic activity of MMP2 was analyzed by using gelatin zymography. Fibroblast migration was evaluated by using transwell migration assays. Contractile activity was measured by a collagen gel contraction assay. Results: The expression level of MMP2 in nasal polyp tissues increased in comparison with inferior turbinate tissues. TGF-beta-1'induced NPDFs were not affected by doxycycline (0'40 μg/mL). The expression levels of MMP2 and activation of Smad2/3 in TGF-beta-1'induced NPDFs and in organ cultures of nasal polyps were significantly downregulated with doxycycline pretreatment. Doxycycline also reduced TGF-beta-1'induced fibroblast migration and collagen contraction in NPDFs. Conclusion: Doxycycline inhibited TGF-beta-1'induced MMP2 expression, migration, and collagen contraction via the Smad2/3 signal pathways in NPDFs.
UR - http://www.scopus.com/inward/record.url?scp=84995912295&partnerID=8YFLogxK
U2 - 10.2500/ajra.2016.30.4381
DO - 10.2500/ajra.2016.30.4381
M3 - Article
C2 - 28124647
AN - SCOPUS:84995912295
SN - 1945-8924
VL - 30
SP - 385
EP - 390
JO - American Journal of Rhinology and Allergy
JF - American Journal of Rhinology and Allergy
IS - 6
ER -