Journal of Molecular Medicine

, Volume 93, Issue 3, pp 289–302 | Cite as

Angiotensin II stimulates canonical TGF-β signaling pathway through angiotensin type 1 receptor to induce granulation tissue contraction

  • Tosan Ehanire
  • Licheng Ren
  • Jennifer Bond
  • Manuel Medina
  • George Li
  • Latif Bashirov
  • Lei Chen
  • George Kokosis
  • Mohamed Ibrahim
  • Angelica Selim
  • Gerard C. Blobe
  • Howard Levinson
Original Article

Abstract

Hypertrophic scar contraction (HSc) is caused by granulation tissue contraction propagated by myofibroblast and fibroblast migration and contractility. Identifying the stimulants that promote migration and contractility is key to mitigating HSc. Angiotensin II (AngII) promotes migration and contractility of heart, liver, and lung fibroblasts; thus, we investigated the mechanisms of AngII in HSc. Human scar and unwounded dermis were immunostained for AngII receptors angiotensin type 1 receptor (AT1 receptor) and angiotensin type 2 receptor (AT2 receptor) and analyzed for AT1 receptor expression using Western blot. In vitro assays of fibroblast contraction and migration under AngII stimulation were conducted with AT1 receptor, AT2 receptor, p38, Jun N-terminal kinase (JNK), MEK, and activin receptor-like kinase 5 (ALK5) antagonism. Excisional wounds were created on AT1 receptor KO and wild-type (WT) mice treated with AngII ± losartan and ALK5 and JNK inhibitors SB-431542 and SP-600125, respectively. Granulation tissue contraction was quantified, and wounds were analyzed by immunohistochemistry. AT1 receptor expression was increased in scar, but not unwounded tissue. AngII induced fibroblast contraction and migration through AT1 receptor. Cell migration was inhibited by ALK5 and JNK, but not p38 or MEK blockade. In vivo experiments determined that absence of AT1 receptor and chemical AT1 receptor antagonism diminished granulation tissue contraction while AngII stimulated wound contraction. AngII granulation tissue contraction was diminished by ALK5 inhibition, but not JNK. AngII promotes granulation tissue contraction through AT1 receptor and downstream canonical transforming growth factor (TGF)-β signaling pathway, ALK5. Further understanding the pathogenesis of HSc as an integrated signaling mechanism could improve our approach to establishing effective therapeutic interventions.

Key message

  • AT1 receptor expression is increased in scar tissue compared to unwounded tissue.

  • AngII stimulates expression of proteins that confer cell migration and contraction.

  • AngII stimulates fibroblast migration and contraction through AT1 receptor, ALK5, and JNK.

  • AngII-stimulated in vivo granulation tissue contraction is AT1 receptor and ALK5 dependent.

Keywords

Angiotensin II AT1 receptor Hypertrophic scar contraction ALK 5 

References

  1. 1.
    ReSurge (2009) The forgotten global health crisis of burnsGoogle Scholar
  2. 2.
    Wang R, Ghahary A, Shen Q, Scott PG, Roy K, Tredget EE (2000) Hypertrophic scar tissues and fibroblasts produce more transforming growth factor-beta1 mRNA and protein than normal skin and cells. Wound Repair Regen: Off Publ Wound Healing Soc Eur Tissue Repair Soc 8:128–137CrossRefGoogle Scholar
  3. 3.
    Zhu Z, Ding J, Shankowsky HA, Tredget EE (2013) The molecular mechanism of hypertrophic scar. J Cell Commun Signal. doi:10.1007/s12079-013-0195-5 PubMedCentralPubMedGoogle Scholar
  4. 4.
    Clayton JL, Edkins R, Cairns BA, Hultman CS (2013) Handle with care: incidence and management of adverse events after the use of laser therapies for the treatment of hypertrophic burn scars. Ann Plast Surg. doi:10.1097/SAP.0b013e31827eac79 PubMedGoogle Scholar
  5. 5.
    Aarabi S, Longaker MT, Gurtner GC (2007) Hypertrophic scar formation following burns and trauma: new approaches to treatment. PLoS Med 4:e234CrossRefPubMedCentralPubMedGoogle Scholar
  6. 6.
    Fearmonti RM, Bond JE, Erdmann D, Levin LS, Pizzo SV, Levinson H (2011) The modified Patient and Observer Scar Assessment Scale: a novel approach to defining pathologic and nonpathologic scarring. Plast Reconstr Surg 127:242–247CrossRefPubMedCentralPubMedGoogle Scholar
  7. 7.
    Washio H, Fukuda N, Matsuda H, Nagase H, Watanabe T, Matsumoto Y, Terui T (2011) Transcriptional inhibition of hypertrophic scars by a gene silencer, pyrrole-imidazole polyamide, targeting the TGF-beta1 promoter. J Investig Dermatol 131:1987–1995CrossRefPubMedGoogle Scholar
  8. 8.
    Kondo S, Kagami S, Urushihara M, Kitamura A, Shimizu M, Strutz F, Muller GA, Kuroda Y (2004) Transforming growth factor-beta1 stimulates collagen matrix remodeling through increased adhesive and contractive potential by human renal fibroblasts. Biochim Biophys Acta 1693:91–100CrossRefPubMedGoogle Scholar
  9. 9.
    Vicente-Manzanares M, Ma X, Adelstein RS, Horwitz AR (2009) Non-muscle myosin II takes centre stage in cell adhesion and migration. Nat Rev Mol Cell Biol 10:778–790CrossRefPubMedCentralPubMedGoogle Scholar
  10. 10.
    Cohn RD, van Erp C, Habashi JP, Soleimani AA, Klein EC, Lisi MT, Gamradt M, ap Rhys CM, Holm TM, Loeys BL et al (2007) Angiotensin II type 1 receptor blockade attenuates TGF-beta-induced failure of muscle regeneration in multiple myopathic states. Nat Med 13:204–210CrossRefPubMedCentralPubMedGoogle Scholar
  11. 11.
    Li L, Fan D, Wang C, Wang JY, Cui XB, Wu D, Zhou Y, Wu LL (2011) Angiotensin II increases periostin expression via Ras/p38 MAPK/CREB and ERK1/2/TGF-beta1 pathways in cardiac fibroblasts. Cardiovasc Res 91:80–89CrossRefPubMedGoogle Scholar
  12. 12.
    Xia CY, Li L, Liu HM, Cong WM (2009) High expression of angiotensin-converting enzyme and angiotensin-converting enzyme 2 in preservation injury after liver transplantation in rats. Hepatol Res: Off J Jpn Soc Hepatol 39:1118–1124CrossRefGoogle Scholar
  13. 13.
    Liu L, Qiu HB, Yang Y, Wang L, Ding HM, Li HP (2009) Losartan, an antagonist of AT1 receptor for angiotensin II, attenuates lipopolysaccharide-induced acute lung injury in rat. Arch Biochem Biophys 481:131–136CrossRefPubMedGoogle Scholar
  14. 14.
    Morihara K, Takai S, Takenaka H, Sakaguchi M, Okamoto Y, Morihara T, Miyazaki M, Kishimoto S (2006) Cutaneous tissue angiotensin-converting enzyme may participate in pathologic scar formation in human skin. J Am Acad Dermatol 54:251–257CrossRefPubMedGoogle Scholar
  15. 15.
    Uzun H, Bitik O, Hekimoglu R, Atilla P, Kayikcioglu AU (2013) Angiotensin-converting enzyme inhibitor enalapril reduces formation of hypertrophic scars in a rabbit ear wounding model. Plast Reconstr Surg 132:361e–371eCrossRefPubMedGoogle Scholar
  16. 16.
    Varkey M, Ding J, Tredget EE (2011) Differential collagen-glycosaminoglycan matrix remodeling by superficial and deep dermal fibroblasts: potential therapeutic targets for hypertrophic scar. Biomaterials 32:7581–7591CrossRefPubMedGoogle Scholar
  17. 17.
    Iannello S, Milazzo P, Bordonaro F, Belfiore F (2006) Low-dose enalapril in the treatment of surgical cutaneous hypertrophic scar and keloid—two case reports and literature review. Med Gen Med 8:60Google Scholar
  18. 18.
    Liu HW, Cheng B, Yu WL, Sun RX, Zeng D, Wang J, Liao YX, Fu XB (2006) Angiotensin II regulates phosphoinositide 3 kinase/Akt cascade via a negative crosstalk between AT1 and AT2 receptors in skin fibroblasts of human hypertrophic scars. Life Sci 79:475–483CrossRefPubMedGoogle Scholar
  19. 19.
    Steckelings UM, Henz BM, Wiehstutz S, Unger T, Artuc M (2005) Differential expression of angiotensin receptors in human cutaneous wound healing. Br J Dermatol 153:887–893CrossRefPubMedGoogle Scholar
  20. 20.
    de la Cruz-Merino L, Henao-Carrasco F, Garcia-Manrique T, Fernandez-Salguero PM, Codes-Manuel de Villena M (2009) Role of transforming growth factor beta in cancer microenvironment. Clin Transl Oncol 11:715–720CrossRefPubMedGoogle Scholar
  21. 21.
    Zhang YE (2009) Non-Smad pathways in TGF-beta signaling. Cell Res 19:128–139CrossRefPubMedCentralPubMedGoogle Scholar
  22. 22.
    Bond JE, Bergeron A, Thurlow P, Selim MA, Bowers EV, Kuang A, Levinson H (2011) Angiotensin-II mediates nonmuscle myosin II activation and expression and contributes to human keloid disease progression. Mol Med 17:1196–1203CrossRefPubMedCentralPubMedGoogle Scholar
  23. 23.
    Jadhav SS, Sharma N, Meeks CJ, Mordwinkin NM, Espinoza TB, Roda NR, Dizerega GS, Hill CK, Louie SG, Rodgers KE (2013) Effects of combined radiation and burn injury on the renin-angiotensin system. Wound Repair Regen: Off Publ Wound Healing Soc Eur Tissue Repair Soc 21:131–140CrossRefGoogle Scholar
  24. 24.
    Takeda H, Katagata Y, Hozumi Y, Kondo S (2004) Effects of angiotensin II receptor signaling during skin wound healing. Am J Pathol 165:1653–1662CrossRefPubMedCentralPubMedGoogle Scholar
  25. 25.
    Ehrlich HP, Rajaratnam JB (1990) Cell locomotion forces versus cell contraction forces for collagen lattice contraction: an in vitro model of wound contraction. Tissue Cell 22:407–417CrossRefPubMedGoogle Scholar
  26. 26.
    Tomasek JJ, Vaughan MB, Kropp BP, Gabbiani G, Martin MD, Haaksma CJ, Hinz B (2006) Contraction of myofibroblasts in granulation tissue is dependent on Rho/Rho kinase/myosin light chain phosphatase activity. Wound Repair Regen: Off Publ Wound Healing Soc Eur Tissue Repair Soc 14:313–320CrossRefGoogle Scholar
  27. 27.
    Ramachandran A, Gangopadhyay SS, Krishnan R, Ranpura SA, Rajendran K, Ram-Mohan S, Mulone M, Gong EM, Adam RM (2013) JunB mediates basal- and TGFbeta1-induced smooth muscle cell contractility. PLoS One 8:e53430CrossRefPubMedCentralPubMedGoogle Scholar
  28. 28.
    Diegelmann RF, Evans MC (2004) Wound healing: an overview of acute, fibrotic and delayed healing. Front Biosci 9:283–289CrossRefPubMedGoogle Scholar
  29. 29.
    Tomasek JJ, Gabbiani G, Hinz B, Chaponnier C, Brown RA (2002) Myofibroblasts and mechano-regulation of connective tissue remodelling. Nat Rev Mol Cell Biol 3:349–363CrossRefPubMedGoogle Scholar
  30. 30.
    Trabucchi E, Radaelli E, Marazzi M, Foschi D, Musazzi M, Veronesi AM, Montorsi W (1988) The role of mast cells in wound healing. Int J Tissue React 10:367–372PubMedGoogle Scholar
  31. 31.
    Dong X, Geng Z, Zhao Y, Chen J, Cen Y (2013) Involvement of mast cell chymase in burn wound healing in hamsters. Exp Ther Med 5:643–647PubMedCentralPubMedGoogle Scholar
  32. 32.
    Martin MM, Buckenberger JA, Jiang J, Malana GE, Knoell DL, Feldman DS, Elton TS (2007) TGF-beta1 stimulates human AT1 receptor expression in lung fibroblasts by cross talk between the Smad, p38 MAPK, JNK, and PI3K signaling pathways. Am J Physiol Lung Cell Mol Physiol 293:L790–L799CrossRefPubMedCentralPubMedGoogle Scholar
  33. 33.
    van Beuge MM, Prakash J, Lacombe M, Post E, Reker-Smit C, Beljaars L, Poelstra K (2013) Enhanced effectivity of an ALK5-inhibitor after cell-specific delivery to hepatic stellate cells in mice with liver injury. PLoS One 8:e56442CrossRefPubMedCentralPubMedGoogle Scholar
  34. 34.
    Higashiyama H, Yoshimoto D, Kaise T, Matsubara S, Fujiwara M, Kikkawa H, Asano S, Kinoshita M (2007) Inhibition of activin receptor-like kinase 5 attenuates bleomycin-induced pulmonary fibrosis. Exp Mol Pathol 83:39–46CrossRefPubMedGoogle Scholar
  35. 35.
    Reish RG, Eriksson E (2008) Scar treatments: preclinical and clinical studies. J Am Coll Surg 206:719–730CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2014

Authors and Affiliations

  • Tosan Ehanire
    • 1
    • 3
  • Licheng Ren
    • 2
    • 3
  • Jennifer Bond
    • 3
  • Manuel Medina
    • 3
  • George Li
    • 1
    • 4
    • 5
  • Latif Bashirov
    • 3
  • Lei Chen
    • 3
    • 6
  • George Kokosis
    • 3
  • Mohamed Ibrahim
    • 3
  • Angelica Selim
    • 7
  • Gerard C. Blobe
    • 4
    • 5
  • Howard Levinson
    • 3
    • 7
  1. 1.Duke University School of MedicineDuke University Medical Center (DUMC)DurhamUSA
  2. 2.First Affiliated Hospital of Sun Yat-sen UniversityGuangzhouPeople’s Republic of China
  3. 3.Division of Plastic and Reconstructive Surgery, Department of SurgeryDUMCDurhamUSA
  4. 4.Department of MedicineDUMCDurhamUSA
  5. 5.Department of Pharmacology and Cancer BiologyDUMCDurhamUSA
  6. 6.Xiangya Hospital, Central South UniversityChangshaPeople’s Republic of China
  7. 7.Department of PathologyDUMCDurhamUSA

Personalised recommendations