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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
Article

Abstract

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.

Keywords

Smad7 Heme oxygenase-1 ROS MMP9 Cardiac fibrosis 

Abbreviations

AngII

AngiotensinII

ARBs

Angiotensin II type 1 receptor blockers

AF

Atrial fibrillation

MI

Myocardial infarct

MMPs

Matrix metalloproteinases

EMT

Epithelial-to-mesenchymal transition

SHRs

Spontaneously hypertensive rats

ROS

Reactive oxygen species

HO-1

Heme oxygenase-1

SOD

Superoxide dismutase

Gpx1

Glutathione peroxidase

Prdx2

Peroxiredoxin-2

CO

Carbon monoxide

FBS

Fetal bovine serum

siRNA

Small interfering RNA

Notes

Acknowledgments

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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