Molecular and Cellular Biochemistry

, Volume 365, Issue 1–2, pp 181–189 | Cite as

Neuregulin induces CTGF expression in hypertrophic scarring fibroblasts

  • Jun-Sub Kim
  • Ihn-Geun Choi
  • Boung-Chul Lee
  • Jae-Bong Park
  • Jin-Hee Kim
  • Je Hoon Jeong
  • Ji Hoon Jeong
  • Cheong Hoon Seo
Article
First Online:
Received:
Accepted:

Abstract

Hypertrophic scarring (HTS) is a common fibroproliferative disorder that typically follows thermal and other injuries involving the deep dermis. These pathogenic mechanisms are regulated by connective tissue growth factor (CTGF) and transforming growth factor-β. We found that neuregulin-1 (NRG1), as well as NRG receptors, HER-2, and HER-3 were upregulated in HTS fibroblasts (HTSF), compared with normal fibroblasts. Furthermore, NRG1 stimulation increased the expression of CTGF in HTSF. In the presence of inhibitors of PI3K, Src, Smad, or reactive oxygen species, the effect of NRG1 on CTGF expression decreased significantly. In particular, the combination of LY294002 or PP2 with SB431542 blocked NRG1-mediated CTGF expression in HTSF. Finally, we demonstrated that siRNA for CTGF, AG825, LY294002, and PP2, either alone or in co-treatment, effectively reduced extracellular matrix expression. Taken together, our results suggest that NRG1 is involved in fibrotic scar pathogenesis via PI3K- or Src-mediated CTGF expression.

Keywords

Neuregulin CTGF Scar Burn 

Abbreviations

HTS

Hypertrophic scarring

CTGF

Connective tissue growth factor

TGF-β

Transforming growth factor-β

NRG

Neuregulin

α-SMA

α-Smooth muscle actin

ROS

Reactive oxygen species

ECM

Extracellular matrix

GAPDH

Glyceraldehyde-3-phosphate dehydrogenase

COL-I

Collagen type I

Nox

NADPH-oxidase

This is a preview of subscription content, log in to check access.

Notes

Acknowledgments

We acknowledge Ji-Yeon Seo for comments and criticisms. These studies were supported by the Korea Healthcare technology R&D Project, Ministry for Health, Welfare and Family Affairs, Republic of Korea (A084589) and the National Research Foundation of Korea (NRF-2010-013-E00019).

Conflicts of interest

None.

References

  1. 1.
    Scott PG, Ghahary A, Tredget EE (2000) Molecular and cellular aspects of fibrosis following thermal injury. Hand Clin 16:271–287PubMedGoogle Scholar
  2. 2.
    Bellemare J, Roberge CJ, Bergeron D, Lopez-Valle CA, Roy M, Moulin VJ (2005) Epidermis promotes dermal fibrosis: role in the pathogenesis of hypertrophic scars. J Pathol 206:1–8PubMedCrossRefGoogle Scholar
  3. 3.
    Frazier K, Williams S, Kothapalli D, Klapper H, Grotendorst GR (1996) Stimulation of fibroblast cell growth, matrix production, and granulation tissue formation by connective tissue growth factor. J Invest Dermatol 107:404–411PubMedCrossRefGoogle Scholar
  4. 4.
    Leask A, Denton CP, Abraham DJ (2004) Insights into the molecular mechanism of chronic fibrosis: the role of connective tissue growth factor in scleroderma. J Invest Dermatol 122:1–6PubMedCrossRefGoogle Scholar
  5. 5.
    Lin CG, Chen CC, Leu SJ, Grzeszkiewicz TM, Lau LF (2005) Integrin-dependent functions of the angiogenic inducer NOV (CCN3): implication in wound healing. J Biol Chem 280:8229–8237PubMedCrossRefGoogle Scholar
  6. 6.
    Igarashi A, Okochi H, Bradham DM, Grotendorst GR (1993) Regulation of connective tissue growth factor gene expression in human skin fibroblasts and during wound repair. Mol Biol Cell 4:637–645PubMedGoogle Scholar
  7. 7.
    Dammeier J, Beer HD, Brauchle M, Werner S (1998) Dexamethasone is a novel potent inducer of connective tissue growth factor expression. Implications for glucocorticoid therapy. J Biol Chem 273:18185–18190PubMedCrossRefGoogle Scholar
  8. 8.
    Igarashi A, Nashiro K, Kikuchi K, Sato S, Ihn H, Fujimoto M, Grotendorst GR, Takehara K (1996) Connective tissue growth factor gene expression in tissue sections from localized scleroderma, keloid, and other fibrotic skin disorders. J Invest Dermatol 106:729–733PubMedCrossRefGoogle Scholar
  9. 9.
    Colwell AS, Phan TT, Kong W, Longaker MT, Lorenz PH (2005) Hypertrophic scar fibroblasts have increased connective tissue growth factor expression after transforming growth factor-beta stimulation. Plast Reconstr Surg 116:1387–1390 discussion 1391–1382PubMedCrossRefGoogle Scholar
  10. 10.
    Shi-wen X, Pennington D, Holmes A, Leask A, Bradham D, Beauchamp JR, Fonseca C, du Bois RM, Martin GR, Black CM, Abraham DJ (2000) Autocrine overexpression of CTGF maintains fibrosis: RDA analysis of fibrosis genes in systemic sclerosis. Exp Cell Res 259:213–224PubMedCrossRefGoogle Scholar
  11. 11.
    Blalock TD, Duncan MR, Varela JC, Goldstein MH, Tuli SS, Grotendorst GR, Schultz GS (2003) Connective tissue growth factor expression and action in human corneal fibroblast cultures and rat corneas after photorefractive keratectomy. Invest Ophthalmol Vis Sci 44:1879–1887PubMedCrossRefGoogle Scholar
  12. 12.
    Kessler D, Dethlefsen S, Haase I, Plomann M, Hirche F, Krieg T, Eckes B (2001) Fibroblasts in mechanically stressed collagen lattices assume a “synthetic” phenotype. J Biol Chem 276:36575–36585PubMedCrossRefGoogle Scholar
  13. 13.
    Park SK, Kim J, Seomun Y, Choi J, Kim DH, Han IO, Lee EH, Chung SK, Joo CK (2001) Hydrogen peroxide is a novel inducer of connective tissue growth factor. Biochem Biophys Res Commun 284:966–971PubMedCrossRefGoogle Scholar
  14. 14.
    Xu SW, Howat SL, Renzoni EA, Holmes A, Pearson JD, Dashwood MR, Bou-Gharios G, Denton CP, du Bois RM, Black CM, Leask A, Abraham DJ (2004) Endothelin-1 induces expression of matrix-associated genes in lung fibroblasts through MEK/ERK. J Biol Chem 279:23098–23103PubMedCrossRefGoogle Scholar
  15. 15.
    Brigstock DR (2002) Regulation of angiogenesis and endothelial cell function by connective tissue growth factor (CTGF) and cysteine-rich 61 (CYR61). Angiogenesis 5:153–165PubMedCrossRefGoogle Scholar
  16. 16.
    Suzuma K, Naruse K, Suzuma I, Takahara N, Ueki K, Aiello LP, King GL (2000) Vascular endothelial growth factor induces expression of connective tissue growth factor via KDR, Flt1, and phosphatidylinositol 3-kinase-akt-dependent pathways in retinal vascular cells. J Biol Chem 275:40725–40731PubMedCrossRefGoogle Scholar
  17. 17.
    Bradham DM, Igarashi A, Potter RL, Grotendorst GR (1991) Connective tissue growth factor: a cysteine-rich mitogen secreted by human vascular endothelial cells is related to the SRC-induced immediate early gene product CEF-10. J Cell Biol 114:1285–1294PubMedCrossRefGoogle Scholar
  18. 18.
    Wenger C, Ellenrieder V, Alber B, Lacher U, Menke A, Hameister H, Wilda M, Iwamura T, Beger HG, Adler G, Gress TM (1999) Expression and differential regulation of connective tissue growth factor in pancreatic cancer cells. Oncogene 18:1073–1080PubMedCrossRefGoogle Scholar
  19. 19.
    Olayioye MA, Neve RM, Lane HA, Hynes NE (2000) The ErbB signaling network: receptor heterodimerization in development and cancer. EMBO J 19:3159–3167PubMedCrossRefGoogle Scholar
  20. 20.
    Yarden Y, Sliwkowski MX (2001) Untangling the ErbB signalling network. Natl Rev Mol Cell Biol 2:127–137CrossRefGoogle Scholar
  21. 21.
    Mei L, Xiong WC (2008) Neuregulin 1 in neural development, synaptic plasticity and schizophrenia. Nat Rev Neurosci 9:437–452PubMedCrossRefGoogle Scholar
  22. 22.
    Burgess AW, Cho HS, Eigenbrot C, Ferguson KM, Garrett TP, Leahy DJ, Lemmon MA, Sliwkowski MX, Ward CW, Yokoyama S (2003) An open-and-shut case? Recent insights into the activation of EGF/ErbB receptors. Mol Cell 12:541–552PubMedCrossRefGoogle Scholar
  23. 23.
    Stein RA, Staros JV (2006) Insights into the evolution of the ErbB receptor family and their ligands from sequence analysis. BMC Evol Biol 6:79PubMedCrossRefGoogle Scholar
  24. 24.
    Marikovsky M, Lavi S, Pinkas-Kramarski R, Karunagaran D, Liu N, Wen D, Yarden Y (1995) ErbB-3 mediates differential mitogenic effects of NDF/heregulin isoforms on mouse keratinocytes. Oncogene 10:1403–1411PubMedGoogle Scholar
  25. 25.
    De Potter IY, Poumay Y, Squillace KA, Pittelkow MR (2001) Human EGF receptor (HER) family and heregulin members are differentially expressed in epidermal keratinocytes and modulate differentiation. Exp Cell Res 271:315–328PubMedCrossRefGoogle Scholar
  26. 26.
    Schelfhout VR, Coene ED, Delaey B, Waeytens AA, De Rycke L, Deleu M, De Potter CR (2002) The role of heregulin-alpha as a motility factor and amphiregulin as a growth factor in wound healing. J Pathol 198:523–533PubMedCrossRefGoogle Scholar
  27. 27.
    Choi W, Wolber R, Gerwat W, Mann T, Batzer J, Smuda C, Liu H, Kolbe L, Hearing VJ (2010) The fibroblast-derived paracrine factor neuregulin-1 has a novel role in regulating the constitutive color and melanocyte function in human skin. J Cell Sci 123:3102–3111PubMedCrossRefGoogle Scholar
  28. 28.
    Kim SH, Jeong JH, Lee SH, Kim SW, Park TG (2006) PEG conjugated VEGF siRNA for anti-angiogenic gene therapy. J Control Release 116:123–129PubMedCrossRefGoogle Scholar
  29. 29.
    Kim SH, Jeong JH, Lee SH, Kim SW, Park TG (2008) Local and systemic delivery of VEGF siRNA using polyelectrolyte complex micelles for effective treatment of cancer. J Control Release 129:107–116PubMedCrossRefGoogle Scholar
  30. 30.
    Wang J, Dodd C, Shankowsky HA, Scott PG, Tredget EE (2008) Deep dermal fibroblasts contribute to hypertrophic scarring. Lab Invest 88:1278–1290PubMedCrossRefGoogle Scholar
  31. 31.
    Kim JS, Bak EJ, Lee BC, Kim YS, Park JB, Choi IG (2011) Neuregulin induces HaCaT keratinocyte migration via Rac1-mediated NADPH-oxidase activation. J Cell Physiol 226:3014–3021PubMedCrossRefGoogle Scholar
  32. 32.
    Hasegawa T, Nakao A, Sumiyoshi K, Tsuchihashi H, Ogawa H (2005) SB-431542 inhibits TGF-beta-induced contraction of collagen gel by normal and keloid fibroblasts. J Dermatol Sci 39:33–38PubMedCrossRefGoogle Scholar
  33. 33.
    Graness A, Cicha I, Goppelt-Struebe M (2006) Contribution of Src-FAK signaling to the induction of connective tissue growth factor in renal fibroblasts. Kidney Int 69:1341–1349PubMedGoogle Scholar
  34. 34.
    Samarin J, Cicha I, Goppelt-Struebe M (2009) Cell type-specific regulation of CCN2 protein expression by PI3K-AKT-FoxO signaling. J Cell Commun Signal 3:79–84PubMedCrossRefGoogle Scholar
  35. 35.
    Chalazonitis A, D’Autreaux F, Pham TD, Kessler JA, Gershon MD (2011) Bone morphogenetic proteins regulate enteric gliogenesis by modulating ErbB3 signaling. Dev Biol 350:64–79PubMedCrossRefGoogle Scholar
  36. 36.
    Sepp-Lorenzino L, Eberhard I, Ma Z, Cho C, Serve H, Liu F, Rosen N, Lupu R (1996) Signal transduction pathways induced by heregulin in MDA-MB-453 breast cancer cells. Oncogene 12:1679–1687PubMedGoogle Scholar
  37. 37.
    Yokoi H, Mukoyama M, Nagae T, Mori K, Suganami T, Sawai K, Yoshioka T, Koshikawa M, Nishida T, Takigawa M, Sugawara A, Nakao K (2004) Reduction in connective tissue growth factor by antisense treatment ameliorates renal tubulointerstitial fibrosis. J Am Soc Nephrol 15:1430–1440PubMedCrossRefGoogle Scholar
  38. 38.
    Al-Refu K, Goodfield M (2009) Scar classification in cutaneous lupus erythematosus: morphological description. Br J Dermatol 161:1052–1058PubMedCrossRefGoogle Scholar
  39. 39.
    Cucoranu I, Clempus R, Dikalova A, Phelan PJ, Ariyan S, Dikalov S, Sorescu D (2005) NAD(P)H oxidase 4 mediates transforming growth factor-beta1-induced differentiation of cardiac fibroblasts into myofibroblasts. Circ Res 97:900–907PubMedCrossRefGoogle Scholar
  40. 40.
    Goldsmit Y, Erlich S, Pinkas-Kramarski R (2001) Neuregulin induces sustained reactive oxygen species generation to mediate neuronal differentiation. Cell Mol Neurobiol 21:753–769PubMedCrossRefGoogle Scholar
  41. 41.
    Mori T, Kawara S, Shinozaki M, Hayashi N, Kakinuma T, Igarashi A, Takigawa M, Nakanishi T, Takehara K (1999) Role and interaction of connective tissue growth factor with transforming growth factor-beta in persistent fibrosis: a mouse fibrosis model. J Cell Physiol 181:153–159PubMedCrossRefGoogle Scholar
  42. 42.
    Bonniaud P, Martin G, Margetts PJ, Ask K, Robertson J, Gauldie J, Kolb M (2004) Connective tissue growth factor is crucial to inducing a profibrotic environment in “fibrosis-resistant” BALB/c mouse lungs. Am J Respir Cell Mol Biol 31:510–516PubMedCrossRefGoogle Scholar
  43. 43.
    Sonnylal S, Shi-Wen X, Leoni P, Naff K, Van Pelt CS, Nakamura H, Leask A, Abraham D, Bou-Gharios G, de Crombrugghe B (2010) Selective expression of connective tissue growth factor in fibroblasts in vivo promotes systemic tissue fibrosis. Arthritis Rheum 62:1523–1532PubMedCrossRefGoogle Scholar
  44. 44.
    Shi-wen X, Stanton LA, Kennedy L, Pala D, Chen Y, Howat SL, Renzoni EA, Carter DE, Bou-Gharios G, Stratton RJ, Pearson JD, Beier F, Lyons KM, Black CM, Abraham DJ, Leask A (2006) CCN2 is necessary for adhesive responses to transforming growth factor-beta1 in embryonic fibroblasts. J Biol Chem 281:10715–10726PubMedCrossRefGoogle Scholar
  45. 45.
    Ponticos M, Holmes AM, Shi-wen X, Leoni P, Khan K, Rajkumar VS, Hoyles RK, Bou-Gharios G, Black CM, Denton CP, Abraham DJ, Leask A, Lindahl GE (2009) Pivotal role of connective tissue growth factor in lung fibrosis: MAPK-dependent transcriptional activation of type I collagen. Arthritis Rheum 60:2142–2155PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC. 2012

Authors and Affiliations

  • Jun-Sub Kim
    • 1
  • Ihn-Geun Choi
    • 2
  • Boung-Chul Lee
    • 2
  • Jae-Bong Park
    • 3
  • Jin-Hee Kim
    • 1
  • Je Hoon Jeong
    • 4
  • Ji Hoon Jeong
    • 5
  • Cheong Hoon Seo
    • 6
  1. 1.Specialization Research Center, Hallym University Burn InstituteHangang Sacred Heart Hospital, Hallym UniversitySeoulSouth Korea
  2. 2.Department of Neuropsychiatry, Hallym University Burn InstituteHangang Sacred Heart Hospital, Hallym UniversitySeoulSouth Korea
  3. 3.Department of Biochemistry, Hallym University Burn InstituteCollege of Medicine, Hallym UniversityChuncheonSouth Korea
  4. 4.Department of Neurological Surgery, Hallym University Burn InstituteHangang Sacred Heart Hospital, Hallym UniversitySeoulSouth Korea
  5. 5.College of PharmacySungkyunkwan UniversitySuwonSouth Korea
  6. 6.Department of Rehabilitation MedicineHangang Sacred Heart Hospital, Hallym UniversitySeoulSouth Korea

Personalised recommendations