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N-Acetyl–seryl–aspartyl–lysyl–proline inhibits ET-1-induced collagen production by preserving Src homology 2-containing protein tyrosine phosphatase-2 activity in cardiac fibroblasts

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Abstract

N–Acetyl–seryl–aspartyl–lysyl–proline (Ac-SDKP) inhibits endothelin-1 (ET-1)-induced activation of p44/42 mitogen-activated protein kinase (p44/42 MAPK) and collagen production in cultured rat cardiac fibroblasts (RCFs). However, we do not know whether its inhibitory effect on p44/42 MAPK is due to the altered activity of protein tyrosine phosphatases (PTPs), which in turn downregulate the p44/42 MAPK signaling pathway. The activity of Src homology 2-containing protein tyrosine phosphatase-2 (SHP-2) is downregulated by ET-1 in RCFs; thus, we hypothesized that Ac-SDKP inhibits ET-1-stimulated collagen production in part by preserving SHP-2 activity and thereby inhibiting p44/42 MAPK phosphorylation. When we stimulated RCFs with ET-1 in the presence or absence of Ac-SDKP, we found that (a) PTP activity was reduced by ET-1 and (b) this effect was counteracted by Ac-SDKP in a dose-dependent fashion. Next, we extracted SHP-2 from RCF lysates by immunoprecipitation and determined that (a) ET-1 inhibited SHP-2 by 40 % and (b) this effect was prevented by Ac-SDKP. However, Ac-SDKP failed to inhibit ET-1-induced p44/42 MAPK phosphorylation in RCFs treated with SHP-2 short hairpin RNA (shRNA); in contrast, in cells transfected with control shRNA, Ac-SDKP’s inhibitory effect on ET-1-induced p44/42 MAPK activation remained intact. Moreover, the inhibitory effect of Ac-SDKP on ET-1-stimulated collagen production was blunted in cells treated with the SHP-1/2 inhibitor NSC-87877. Thus, we concluded that the inhibitory effect of Ac-SDKP on ET-1-stimulated collagen production by RCFs is mediated in part by preserving SHP-2 activity and thereby preventing p44/42 MAPK activation. Ac-SDKP or its analogs could represent a new therapeutic tool to treat fibrotic diseases in the cardiovascular system.

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Acknowledgments

This study was supported by National Institutes of Health grants HL 071806 (NER) and HL 028982 (OAC).

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Authors and Affiliations

  1. Hypertension and Vascular Research Division, Department of Internal Medicine, Henry Ford Hospital, E & R Bldg 7121, 2799 West Grand Blvd, Detroit, MI, USA

    Hongmei Peng, Oscar A. Carretero, Xiao-Ping Yang, Kastuv Santra & Nour-Eddine Rhaleb

  2. Department of Physiology, Wayne State University, Detroit, MI, USA

    Nour-Eddine Rhaleb

  3. Department of Biostatistics and Research Epidemiology, Henry Ford Hospital, Detroit, MI, USA

    Edward L. Peterson

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  1. Hongmei Peng
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Correspondence to Nour-Eddine Rhaleb.

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Peng, H., Carretero, O.A., Peterson, E.L. et al. N-Acetyl–seryl–aspartyl–lysyl–proline inhibits ET-1-induced collagen production by preserving Src homology 2-containing protein tyrosine phosphatase-2 activity in cardiac fibroblasts. Pflugers Arch - Eur J Physiol 464, 415–423 (2012). https://doi.org/10.1007/s00424-012-1150-7

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  • DOI: https://doi.org/10.1007/s00424-012-1150-7

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