Molecular Biology Reports

, Volume 40, Issue 9, pp 5307–5314 | Cite as

Overexpression of Smad7 suppressed ROS/MMP9-dependent collagen synthesis through regulation of heme oxygenase-1

  • Hong Yu
  • Junxing Huang
  • Shijun Wang
  • Gang Zhao
  • Xia Jiao
  • Li Zhu


We previously reported that AngiotensinII receptor blocker effectively inhibited TGF-β1-mediated epithelial-to-mesenchymal transition progress through regulating Smad7. However, the underlying mechanism by which Smad7 exerted in regulating MMP9 and fibrogenic response has not been fully elucidated. In the current study, we proved that NADPH p47phox-dependent reactive oxygen species (ROS) production contributed to MMP9 activation and collagen expression, which was suppressed by transfecting pcDNA3–Smad7 in cardiac fibroblasts. The effect of Smad7 overexpression on MMP9 activity and collagen expression was further reversed by adding H2O2 (10 μmol/L). In contrast, knockdown of Smad7 caused the enhanced collagen synthesis in cardiac fibroblasts, which was also reversed by treating cells with a ROS inhibitor, YCG063 (2 μmol/L). Further investigation showed that Smad7 regulated NADPH-mediated ROS production through activating Heme oxygenase-1 (HO-1). Meanwhile, the intercellular level of bilirubin (product of hemin) and nitric oxide (NO) in cell supernatant were not significantly increased in cells treated with AngII or transfected with Smad7. Knockdown of HO-1 in Smad7-overexpressed cardiac fibroblasts or cells pretreated with SnPP IX, a competitive inhibitor of HO-1 activity, resulted in increased productions of ROS and NADPH p47phox, and abolished the inhibitory effects of Smad7 on MMP9 activity and collagen expression. Our results indicated that HO-1 might be critically involved in Smad7-mediated regulation of MMP9 activity and fibrogenic genes expression via antagonizing the enhanced myocardial oxidative stress.


Smad7 Heme oxygenase-1 ROS MMP9 Cardiac fibrosis 





Angiotensin II type 1 receptor blockers


Atrial fibrillation


Myocardial infarct


Matrix metalloproteinases


Epithelial-to-mesenchymal transition


Spontaneously hypertensive rats


Reactive oxygen species


Heme oxygenase-1


Superoxide dismutase


Glutathione peroxidase




Carbon monoxide


Fetal bovine serum


Small interfering RNA



This work was supported by Natural Science Fund programs of Nantong University (10Z088).

Conflict of interest

The authors confirmed that there are no conflicts of interest.

Supplementary material

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Supplementary Table 1 (DOC 33 kb)
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Supplementary Figure 1 (TIFF 2466 kb)
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Supplementary Figure 2 (TIFF 1327 kb)
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Supplementary Figure 3 (TIFF 2628 kb)
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Supplementary Figure 4 (TIFF 1687 kb)


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Copyright information

© Springer Science+Business Media Dordrecht 2013

Authors and Affiliations

  • Hong Yu
    • 2
  • Junxing Huang
    • 3
  • Shijun Wang
    • 4
  • Gang Zhao
    • 4
  • Xia Jiao
    • 2
  • Li Zhu
    • 1
  1. 1.Institute of Cardiovascular DiseasesTaizhou People’s HospitalTaizhouChina
  2. 2.Department of PathologyTaizhou People’s HospitalTaizhouChina
  3. 3.Department of OncologyTaizhou People’s HospitalTaizhouChina
  4. 4.Shanghai Institute of Cardiovascular Diseases, Zhongshan Hospital and Institutes of Biomedical ScienceFudan UniversityShanghaiChina

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