Login Register Cart 0
Generic placeholder image

Current Pharmaceutical Design

Editor-in-Chief

ISSN (Print): 1381-6128
ISSN (Online): 1873-4286

Back
Journal Home About Journal Editorial Board Journal Insight Current Issue Volumes/Issues
Subscribe
Research Article

Thrombin Improves Diabetic Wound Healing by ERK-Dependent and Independent Smad2/3 Linker Region Phosphorylation

Author(s): Gang Luo, Chongyang Wang, Juehong Li, Xuancheng Zhang, Ziyang Sun, Sa Song* and Cunyi Fan*

Volume 28, Issue 17, 2022

Published on: 24 June, 2022

Page: [1433 - 1443] Pages: 11

DOI: 10.2174/1381612828666220511125237

Price: $65

Abstract

Background: Impaired wound healing is one of the most noteworthy features and troublesome complications of diabetes mellitus, which arouses a rising global health concern without potent remedies. Thrombin is the major hemostatic agent applied at wound healing initiation and recently gained therapeutic credits in later phases. However, a rare investigation achieved prolonged use of thrombin and probed the detailed mechanism.

Objective: The objective of this study is to investigate the effects and mechanism of thrombin on diabetic skin wound healing.

Methods: The effect of thrombin on fibroblast proliferation, α-SMA, and Collagen I expression was firstly studied in vitro by Cell Counting Kit 8 (CCK8) and western blotting. Then, the specific phosphorylation site of SMAD2/3 and their ERK1/2 dependence during thrombin treatment were assessed by western blotting for mechanism exploration. After that, full-thickness wound defects were established in diabetic male SD rats and treated with thrombin in the presence or absence of PD98059 to observe the in vivo effects of thrombin and to confirm its ERK dependence.

Results: We found that thrombin promoted fibroblast proliferation and their α-SMA and Collagen I production. Mechanistically, thrombin induced phosphorylation of Smad2 linker region (Ser245/250/255) through ERK1/2 phosphorylation but promoted phosphorylation of Smad3 linker region (Ser204) independent of ERK1/2. Histological results showed that thrombin facilitated wound healing by promoting α-SMA and Collagen I expression, which was not abolished by inhibiting ERK phosphorylation.

Conclusion: Collectively, this study validated the therapeutic efficacy of thrombin on diabetic wound healing and identified both ERK-dependent and -independent Smad2/3 linker region phosphorylation as the essential signaling events in this process.

Keywords: Thrombin, wound healing, Smad2/3, linker region, ERK1/2, hemostatic agent.

« Previous
[1]
Eming SA, Martin P, Tomic-Canic M. Wound repair and regeneration: Mechanisms, signaling, and translation. Sci Transl Med 2014; 6(265): 265sr6.
[http://dx.doi.org/10.1126/scitranslmed.3009337] [PMID: 25473038]
[2]
Kim HS, Sun X, Lee JH, Kim HW, Fu X, Leong KW. Advanced drug delivery systems and artificial skin grafts for skin wound healing. Adv Drug Deliv Rev 2019; 146: 209-39.
[http://dx.doi.org/10.1016/j.addr.2018.12.014] [PMID: 30605737]
[3]
Olczyk P, Mencner Ł Komosinska-Vassev K. The role of the extracellular matrix components in cutaneous wound healing. BioMed Res Int 2014; 2014: 747584.
[http://dx.doi.org/10.1155/2014/747584] [PMID: 24772435]
[4]
Boniakowski AE, Kimball AS, Jacobs BN, Kunkel SL, Gallagher KA. Macrophage-mediated inflammation in normal and diabetic wound healing. J Immunol 2017; 199(1): 17-24.
[http://dx.doi.org/10.4049/jimmunol.1700223] [PMID: 28630109]
[5]
Veith AP, Henderson K, Spencer A, Sligar AD, Baker AB. Therapeutic strategies for enhancing angiogenesis in wound healing. Adv Drug Deliv Rev 2019; 146: 97-125.
[http://dx.doi.org/10.1016/j.addr.2018.09.010] [PMID: 30267742]
[6]
Gugerell A, Pasteiner W, Nürnberger S, et al. Thrombin as important factor for cutaneous wound healing: Comparison of fibrin biomatri-ces in vitro and in a rat excisional wound healing model. Wound Repair Regen 2014; 22(6): 740-8.
[http://dx.doi.org/10.1111/wrr.12234] [PMID: 25231003]
[7]
Ziv-Polat O, Topaz M, Brosh T, Margel S. Enhancement of incisional wound healing by thrombin conjugated iron oxide nanoparticles. Biomaterials 2010; 31(4): 741-7.
[http://dx.doi.org/10.1016/j.biomaterials.2009.09.093] [PMID: 19850336]
[8]
Scherer SS, Tobalem M, Vigato E, et al. Nonactivated versus thrombin-activated platelets on wound healing and fibroblast-to-myofibroblast differentiation in vivo and in vitro. Plast Reconstr Surg 2012; 129(1): 46e-54e.
[http://dx.doi.org/10.1097/PRS.0b013e3182362010] [PMID: 22186584]
[9]
Marin V, Farnarier C, Grès S, et al. The p38 mitogen-activated protein kinase pathway plays a critical role in thrombin-induced endothelial chemokine production and leukocyte recruitment. Blood 2001; 98(3): 667-73.
[http://dx.doi.org/10.1182/blood.V98.3.667] [PMID: 11468165]
[10]
Dolivo DM, Larson SA, Dominko T. Crosstalk between mitogen-activated protein kinase inhibitors and transforming growth factor-β signaling results in variable activation of human dermal fibroblasts. Int J Mol Med 2019; 43(1): 325-35.
[PMID: 30365043]
[11]
Song CY, Kim BC, Lee HS. Lovastatin inhibits oxidized low-density lipoprotein-induced plasminogen activator inhibitor and transform-ing growth factor-beta1 expression via a decrease in Ras/extracellular signal-regulated kinase activity in mesangial cells. Transl Res 2008; 151(1): 27-35.
[http://dx.doi.org/10.1016/j.trsl.2007.09.008] [PMID: 18061125]
[12]
Jiang S, Zhao X, Chen S, et al. Down-regulating ERK1/2 and SMAD2/3 phosphorylation by physical barrier of celecoxib-loaded electrospun fibrous membranes prevents tendon adhesions. Biomaterials 2014; 35(37): 9920-9.
[http://dx.doi.org/10.1016/j.biomaterials.2014.08.028] [PMID: 25201739]
[13]
Li F, Liu S, Ouyang Y, et al. Effect of celecoxib on proliferation, collagen expression, ERK1/2 and SMAD2/3 phosphorylation in NIH/3T3 fibroblasts. Eur J Pharmacol 2012; 678(1-3): 1-5.
[http://dx.doi.org/10.1016/j.ejphar.2011.12.018] [PMID: 22209876]
[14]
Hayashida T, Decaestecker M, Schnaper HW. Cross-talk between ERK MAP kinase and Smad signaling pathways enhances TGF-beta-dependent responses in human mesangial cells. FASEB J 2003; 17(11): 1576-8.
[http://dx.doi.org/10.1096/fj.03-0037fje] [PMID: 12824291]
[15]
Zhu Y, Gu J, Zhu T, Jin C, Hu X, Wang X. Crosstalk between Smad2/3 and specific isoforms of ERK in TGF-β1-induced TIMP-3 expression in rat chondrocytes. J Cell Mol Med 2017; 21(9): 1781-90.
[http://dx.doi.org/10.1111/jcmm.13099] [PMID: 28230313]
[16]
Kong L, Wu Z, Zhao H, et al. Bioactive injectable hydrogels containing desferrioxamine and bioglass for diabetic wound healing. ACS Appl Mater Interfaces 2018; 10(36): 30103-14.
[http://dx.doi.org/10.1021/acsami.8b09191] [PMID: 30113159]
[17]
Carney DH, Mann R, Redin WR, et al. Enhancement of incisional wound healing and neovascularization in normal rats by thrombin and synthetic thrombin receptor-activating peptides. J Clin Invest 1992; 89(5): 1469-77.
[http://dx.doi.org/10.1172/JCI115737] [PMID: 1373740]
[18]
Verkleij CJ, Roelofs JJ, Havik SR, Meijers JC, Marx PF. The role of thrombin-activatable fibrinolysis inhibitor in diabetic wound healing. Thromb Res 2010; 126(5): 442-6.
[http://dx.doi.org/10.1016/j.thromres.2010.08.008] [PMID: 20828799]
[19]
López ML, Bruges G, Crespo G, et al. Thrombin selectively induces transcription of genes in human monocytes involved in inflamma-tion and wound healing. Thromb Haemost 2014; 112(5): 992-1001.
[PMID: 25057055]
[20]
Talati N, Kamato D, Piva TJ, Little PJ, Osman N. Thrombin promotes PAI-1 expression and migration in keratinocytes via ERK dependent Smad linker region phosphorylation. Cell Signal 2018; 47: 37-43.
[http://dx.doi.org/10.1016/j.cellsig.2018.03.009] [PMID: 29577978]
[21]
Yang CC, Hsiao LD, Yang CM, Lin CC. Thrombin enhanced matrix metalloproteinase-9 expression and migration of SK-N-SH Cells via PAR-1, c-Src, PYK2, EGFR, Erk1/2 and AP-1. Mol Neurobiol 2017; 54(5): 3476-91.
[http://dx.doi.org/10.1007/s12035-016-9916-0] [PMID: 27181591]
[22]
Ono M, Masaki A, Maeda A, et al. CCN4/WISP1 controls cutaneous wound healing by modulating proliferation, migration and ECM expression in dermal fibroblasts via α5β1 and TNFα. Matrix Biol 2018; 68-69: 533-46.
[http://dx.doi.org/10.1016/j.matbio.2018.01.004] [PMID: 29330021]
[23]
Alizadeh S, Seyedalipour B, Shafieyan S, Kheime A, Mohammadi P, Aghdami N. Copper nanoparticles promote rapid wound healing in acute full thickness defect via acceleration of skin cell migration, proliferation, and neovascularization. Biochem Biophys Res Commun 2019; 517(4): 684-90.
[http://dx.doi.org/10.1016/j.bbrc.2019.07.110] [PMID: 31400855]
[24]
Wang XT, McKeever CC, Vonu P, Patterson C, Liu PY. Dynamic histological events and molecular changes in excisional wound healing of diabetic DB/DB Mice. J Surg Res 2019; 238: 186-97.
[http://dx.doi.org/10.1016/j.jss.2019.01.048] [PMID: 30771688]
[25]
Ham SA, Hwang JS, Yoo T, et al. Ligand-activated PPARδ upregulates α-smooth muscle actin expression in human dermal fibroblasts: A potential role for PPARδ in wound healing. J Dermatol Sci 2015; 80(3): 186-95.
[http://dx.doi.org/10.1016/j.jdermsci.2015.10.005] [PMID: 26481780]
[26]
Jiang ZT, Li ZZ, Jiang DP. Vanadate affects proliferation of healing fibroblasts, cellular production of collagen, and expression of alpha-SMA in vitro. Med Sci Monit 2010; 16(4): BR107-11.
[PMID: 20357710]
[27]
Somanath PR, Chen J, Byzova TV. Akt1 is necessary for the vascular maturation and angiogenesis during cutaneous wound healing. Angiogenesis 2008; 11(3): 277-88.
[http://dx.doi.org/10.1007/s10456-008-9111-7] [PMID: 18415691]
[28]
Tong M, Tuk B, Hekking IM, Vermeij M, Barritault D, van Neck JW. Stimulated neovascularization, inflammation resolution and collagen maturation in healing rat cutaneous wounds by a heparan sulfate glycosaminoglycan mimetic, OTR4120. Wound Repair Regen 2009; 17(6): 840-52.
[http://dx.doi.org/10.1111/j.1524-475X.2009.00548.x] [PMID: 19903305]
[29]
Sun GF, Li HC, Zhan YP, et al. SnoN residue (1-366) attenuates hypertrophic scars through resistance to transforming growth factor-β1-induced degradation. Lab Invest 2019; 99(12): 1861-73.
[http://dx.doi.org/10.1038/s41374-019-0302-1] [PMID: 31409891]
[30]
Altieri P, Bertolotto M, Fabbi P, et al. Thrombin induces protease-activated receptor 1 signaling and activation of human atrial fibroblasts and dabigatran prevents these effects. Int J Cardiol 2018; 271: 219-27.
[http://dx.doi.org/10.1016/j.ijcard.2018.05.033] [PMID: 29801760]
[31]
Chen YC, Chen BC, Huang HM, Lin SH, Lin CH. Activation of PERK in ET-1- and thrombin-induced pulmonary fibroblast differentiation: Inhibitory effects of curcumin. J Cell Physiol 2019; 234(9): 15977-88.
[http://dx.doi.org/10.1002/jcp.28256] [PMID: 30825198]
[32]
Yao Y, Hu C, Song Q, et al. ADAMTS16 activates latent TGF-β accentuating fibrosis and dysfunction of the pressure-overloaded heart. Cardiovasc Res 2020; 116(5): 956-69.
[http://dx.doi.org/10.1093/cvr/cvz187] [PMID: 31297506]
[33]
Garcia-Rendueles AR, Rodrigues JS, Garcia-Rendueles ME, et al. Rewiring of the apoptotic TGF-β-SMAD/NFκB pathway through an oncogenic function of p27 in human papillary thyroid cancer. Oncogene 2017; 36(5): 652-66.
[http://dx.doi.org/10.1038/onc.2016.233] [PMID: 27452523]
[34]
Lei W, Jian L, Chuanbing H, et al. Synovial and pulmonary dysfunctions are induced by crosstalk of Smad and Erk pathways in an arthritis model. Cell Mol Biol 2020; 66(2): 15-22.
[http://dx.doi.org/10.14715/cmb/2020.66.2.2] [PMID: 32415922]
[35]
Liu L, Li N, Zhang Q, Zhou J, Lin L, He X. Inhibition of ERK1/2 signaling impairs the promoting effects of tgf-β1 on hepatocellular carcinoma cell invasion and epithelial-mesenchymal transition. Oncol Res 2017; 25(9): 1607-16.
[http://dx.doi.org/10.3727/096504017X14938093512742] [PMID: 28492136]
[36]
Sisto M, Lorusso L, Ingravallo G, Ribatti D, Lisi S. TGFβ1-Smad canonical and -Erk noncanonical pathways participate in interleukin-17-induced epithelial-mesenchymal transition in Sjögren’s syndrome. Lab Invest 2020; 100(6): 824-36.
[http://dx.doi.org/10.1038/s41374-020-0373-z] [PMID: 31925325]
[37]
Kamato D, Rostam MA, Piva TJ, et al. Transforming growth factor β-mediated site-specific Smad linker region phosphorylation in vascular endothelial cells. J Pharm Pharmacol 2014; 66(12): 1722-33.
[http://dx.doi.org/10.1111/jphp.12298] [PMID: 25316549]
[38]
Burch ML, Yang SN, Ballinger ML, Getachew R, Osman N, Little PJ. TGF-beta stimulates biglycan synthesis via p38 and ERK phosphorylation of the linker region of Smad2. Cell Mol Life Sci 2010; 67(12): 2077-90.
[http://dx.doi.org/10.1007/s00018-010-0315-9] [PMID: 20213272]
[39]
Hough C, Radu M, Doré JJ. Tgf-beta induced Erk phosphorylation of smad linker region regulates smad signaling. PLoS One 2012; 7(8): e42513.
[http://dx.doi.org/10.1371/journal.pone.0042513] [PMID: 22880011]
[40]
Burzynski LC, Humphry M, Pyrillou K, et al. The coagulation and immune systems are directly linked through the activation of interleukin-1α by thrombin. Immunity 2019; 50(4): 1033-1042.e6.
[http://dx.doi.org/10.1016/j.immuni.2019.03.003] [PMID: 30926232]

Rights & Permissions Print Cite
Article Metrics
24
1
© 2024 Bentham Science Publishers | Privacy Policy