Growth hormone-releasing hormone promotes therapeutic effects of peripheral blood endothelial progenitor cells in ischemic repair
- 25 Downloads
In peripheral artery disease, blockage of the blood supply to the limbs leads to blood flow attenuation and tissue ischemia. We investigated whether growth hormone-releasing hormone (GHRH) could enhance the biological functions and therapeutic effects of endothelial progenitor cells (EPCs) derived from adult human peripheral blood (PB).
EPCs were isolated from human PB (PB-EPCs) and cord blood and expanded in vitro. PB-EPCs incubated with or without GHRH were evaluated for proliferation, migration, and angiogenesis capacity and apoptosis rates under oxidative stress conditions. Activation of STAT3 and Akt pathways was evaluated using Western blot. A hind-limb ischemia (HLI) mouse model was used to study the efficacy of GHRH in improving EPC therapy in vivo.
GHRH enhanced the proliferation, migration, and angiogenesis capacity of PB-EPCs and reduced apoptosis under H2O2 stimulation. These beneficial effects were GHRH receptor-dependent and were paralleled by increased phosphorylation of STAT3 and Akt. Transplantation of GHRH-preconditioned EPCs into HLI model mice enhanced blood flow recovery by increasing vascular formation density and enhanced tissue regeneration at the lesion site.
Our studies demonstrate a novel role for GHRH in dramatically improving therapeutic angiogenesis in HLI by enhancing the biological functions of EPCs. These findings support additional studies to explore the full potential of GHRH in augmenting cell therapy for the management of ischemia.
KeywordsAngiogenesis Endothelial progenitor cell Growth hormone-releasing hormone Hind-limb ischemia Peripheral artery disease
This study was financially supported by Wenzhou Municipal Science and Technology Bureau Foundation (grant # Y20170008 and # Y20170052), Natural Science Foundation of Zhejiang Province (grant # LQ18H090007), and National Natural Science Foundation of China (grant # 81800314).
This study was funded by Wenzhou Municipal Science and Technology Bureau Foundation (grant # Y20170008), Natural Science Foundation of Zhejiang Province (grant # LQ18H090007), Department of Education of Zhejiang Province (Y201636426), and National Natural Science Foundation of China (grant # 81800314).
Compliance with ethical standards
Conflict of interest
The authors have no commercial, proprietary, or financial interest in the products or companies described in this article.
The ethics committee of the First Affiliated Hospital of Wenzhou Medical University approved the study (2017-310).
All informed consents were obtained from all subjects according to the protocol approved by the ethics committee of the First Affiliated Hospital of Wenzhou Medical University (2017-310).
- 2.Selvin E, Erlinger TP (2004) Prevalence of and risk factors for peripheral arterial disease in the United States: results from the National Health and Nutrition Examination Survey, 1999–2000. Circulation 110(6):738–743. https://doi.org/10.1161/01.cir.0000137913.26087.f0 CrossRefGoogle Scholar
- 6.Rauscher FM, Goldschmidt-Clermont PJ, Davis BH, Wang T, Gregg D, Ramaswami P, Pippen AM, Annex BH, Dong C, Taylor DA (2003) Aging, progenitor cell exhaustion, and atherosclerosis. Circulation 108(4):457–463. https://doi.org/10.1161/01.cir.0000082924.75945.48 CrossRefGoogle Scholar
- 7.Mayo KE, Godfrey PA, Suhr ST, Kulik DJ, Rahal JO (1995) Growth hormone-releasing hormone: synthesis and signaling. Recent Prog Horm Res 50:35–73Google Scholar
- 10.Granata R, Trovato L, Gallo MP, Destefanis S, Settanni F, Scarlatti F, Brero A, Ramella R, Volante M, Isgaard J, Levi R, Papotti M, Alloatti G, Ghigo E (2009) Growth hormone-releasing hormone promotes survival of cardiac myocytes in vitro and protects against ischaemia-reperfusion injury in rat heart. Cardiovasc Res 83(2):303–312. https://doi.org/10.1093/cvr/cvp090 CrossRefGoogle Scholar
- 13.Ma Q, Xia X, Tao Q, Lu K, Shen J, Xu Q, Hu X, Tang Y, Block NL, Webster KA, Schally AV, Wang J, Yu H (2016) Profound actions of an agonist of growth hormone-releasing hormone on angiogenic therapy by mesenchymal stem cells. Arterioscler Thromb Vasc Biol 36(4):663–672. https://doi.org/10.1161/ATVBAHA.116.307126 CrossRefGoogle Scholar
- 14.Shen J, Zhang N, Lin YN, Xiang P, Liu XB, Shan PF, Hu XY, Zhu W, Tang YL, Webster KA, Cai R, Schally AV, Wang J, Yu H (2018) Regulation of vascular calcification by growth hormone-releasing hormone and its agonists. Circ Res 122(10):1395–1408. https://doi.org/10.1161/circresaha.117.312418 CrossRefGoogle Scholar
- 15.Russell-Aulet M, Jaffe CA, Demott-Friberg R, Barkan AL (1999) In vivo semiquantification of hypothalamic growth hormone-releasing hormone (GHRH) output in humans: evidence for relative GHRH deficiency in aging. J Clin Endocrinol Metab 84(10):3490–3497. https://doi.org/10.1210/jcem.84.10.6063 Google Scholar
- 18.Thum T, Hoeber S, Froese S, Klink I, Stichtenoth DO, Galuppo P, Jakob M, Tsikas D, Anker SD, Poole-Wilson PA, Borlak J, Ertl G, Bauersachs J (2007) Age-dependent impairment of endothelial progenitor cells is corrected by growth-hormone-mediated increase of insulin-like growth-factor-1. Circ Res 100(3):434–443. https://doi.org/10.1161/01.RES.0000257912.78915.af CrossRefGoogle Scholar
- 19.Fadini GP, Miorin M, Facco M, Bonamico S, Baesso I, Grego F, Menegolo M, de Kreutzenberg SV, Tiengo A, Agostini C, Avogaro A (2005) Circulating endothelial progenitor cells are reduced in peripheral vascular complications of type 2 diabetes mellitus. J Am Coll Cardiol 45(9):1449–1457. https://doi.org/10.1016/j.jacc.2004.11.067 CrossRefGoogle Scholar
- 20.Yuan Q, Hu CP, Gong ZC, Bai YP, Liu SY, Li YJ, Jiang JL (2015) Accelerated onset of senescence of endothelial progenitor cells in patients with type 2 diabetes mellitus: role of dimethylarginine dimethylaminohydrolase 2 and asymmetric dimethylarginine. Biochem Biophys Res Commun 458(4):869–876. https://doi.org/10.1016/j.bbrc.2015.02.050 CrossRefGoogle Scholar
- 21.Bakogiannis C, Tousoulis D, Androulakis E, Briasoulis A, Papageorgiou N, Vogiatzi G, Kampoli AM, Charakida M, Siasos G, Latsios G, Antoniades C, Stefanadis C (2012) Circulating endothelial progenitor cells as biomarkers for prediction of cardiovascular outcomes. Curr Med Chem 19(16):2597–2604CrossRefGoogle Scholar
- 23.Zeng L, Hu Q, Wang X, Mansoor A, Lee J, Feygin J, Zhang G, Suntharalingam P, Boozer S, Mhashilkar A, Panetta CJ, Swingen C, Deans R, From AH, Bache RJ, Verfaillie CM, Zhang J (2007) Bioenergetic and functional consequences of bone marrow-derived multipotent progenitor cell transplantation in hearts with postinfarction left ventricular remodeling. Circulation 115(14):1866–1875. https://doi.org/10.1161/circulationaha.106.659730 CrossRefGoogle Scholar