[AP-1 Decoy modulating MMP-2/TIMP-1 imbalance induced by bleomycin-A5 in pulmonary fibroblasts].
OBJECTIVE
To investigate the effect of AP-1 Decoy on matrix metalloproteinase 2 (MMP-2) and tissue inhibitor of metalloproteinase (TIMP-1) imbalance induced by bleomycin-A5 (BLM-A5) in pulmonary fibroblasts.


METHODS
Pulmonary fibroblasts were primary cultured, and transferred with AP-1 Decoy before treated with BLM-A5. MMPs activity in medium was determined by gelatin zymography. Protein content of TIMP-1 in medium was detected by ELISA. Expression of MMP-2 mRNA and TIMP-1 mRNA were determined by reverse transcriptase-polymerase chain reaction (RT-PCR).


RESULTS
BLM-A5 induced the increase in activity of MMP-2 at 12 h [A: (0.77 +/- 0.08) vs (0.65 +/- 0.07) P < 0.05], but it was suppressed by AP-1 Decoy [A: (0.68 +/- 0.05)]. BLM-A5 up-regulated the expression of protein and mRNA of TIMP-1 after 12 h, and 24 h [(39.3 +/- 4.3), (46.3 +/- 4.8) ng/ml vs (28.9 +/- 2.7), (31.6 +/- 2.4) ng/ml] and [Absorbance ratio to beta-actin: (0.94 +/- 0.13, 1.08 +/- 0.06) vs (0.76 +/- 0.07, 0.75 +/- 0.08)] (P < 0.05 or P < 0.01) but AP-1 Decoy modulated the up-regulation. All these indexes in AP-1 Decoy group had no significant difference in contrast to the normal group. Mutant AP-1 Decoy had not the same function as AP-1 Decoy on the expression of MMP-2 and TIMP-1 in pulmonary fibroblasts.


CONCLUSIONS
AP-1 Decoy inhibits the increase in MMP-2 activity and the up-regulation of TIMP-1 induced by BLM-A5 in pulmonary fibroblasts.
Ma
Wan-li
WL
Department of Respiratory Medicine, Xiehe Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China.
Ye
Hong
H
Xin
Jian-bao
JB
Tao
Xiao-nan
XN
Bai
Ming
M
D001761:Bleomycin / Q000031:analogs & derivatives / Q000494:pharmacology; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D008168:Lung / Q000166:cytology / Q000378:metabolism; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism*; D011658:Pulmonary Fibrosis / Q000378:metabolism; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000378:metabolism*; D018808:Transcription Factor AP-1 / Q000378:metabolism*
C560822:TIMP1 protein, human; D019715:Tissue Inhibitor of Metalloproteinase-1; D018808:Transcription Factor AP-1; D001761:Bleomycin; C025703:bleomycetin; C522361:MMP2 protein, human; D020778:Matrix Metalloproteinase 2