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Bleomycin Induced Imbalance of MMP-2/TIMP-1 in Pulmonary Interstitial Fibroblasts
Wan-Li Ma*, Hong Ye1, Jian-Bao Xin, Xiao-Nan Tao, Ming Bai
Department of Respiratory Medicine, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China; 1Department of Pathophysiology, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 43
Abstract: To investigate the effect of bleomycin on the expression of matrix metalloproteinase-2 (MMP-2) and tissue inhibitor of metalloproteinase-1 (TIMP-1) in pulmonary interstitial fibroblasts. Pulmonary interstitial fibroblasts were primary cultured， then treated with bleomycin-A5. MMP-2 activity in medium was determined by gelatin zymography. Protein content of TIMP-1 in medium was detected by ELISA. Intracellular protein content of MMP-2 and TIMP-1 were detected by immunocytochemistry. Expression of MMP-2 and TIMP-1 mRNA were determined by RT-PCR. The results showed BLM-A5 induced secreting of MMP-2 from pulmonary interstitial fibroblasts at 2 h and 12 h， and there was no effect after 24 h. Intracellular proteins and mRNA of MMP-2 were not influenced by BLM-A5 from 2 h to 48 h. BLM-A5 up-regulated the expression of protein and mRNA of TIMP-1 after 12 h. The results demonstrated that BLM-A5 induced secreting of MMP-2 in early stage in pulmonary interstitial fibroblasts， and then up-regulate the expression of TIMP-1. These maybe contributed to the mechanism of MMP-2/TIMP-1 imbalance in pulmonary fibrosis.
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