Metformin accelerates wound healing in type 2 diabetic db/db mice

Han,Xue; Tao,Yulong; Deng,Yaping; Yu,Jiawen; Sun,Yuannan; Jiang,Guojun

doi:10.3892/mmr.2017.7707

Metformin accelerates wound healing in type 2 diabetic db/db mice

Authors:
- Xue Han
- Yulong Tao
- Yaping Deng
- Jiawen Yu
- Yuannan Sun
- Guojun Jiang
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Affiliations: Department of Pharmacy, Xiaoshan Hospital, Hangzhou, Zhejiang 311202, P.R. China, Department of Pharmacy, Changzheng Hospital, Second Military Medical University, Shanghai 200003, P.R. China
Published online on: October 4, 2017 https://doi.org/10.3892/mmr.2017.7707
Pages: 8691-8698
Copyright: © Han et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Wound healing impairment is increasingly recognized to be a consequence of hyperglycemia‑induced dysfunction of endothelial precursor cells (EPCs) in type 2 diabetes mellitus (T2DM). Metformin exhibits potential for the improvement of endothelial function and the wound healing process. However, the underlying mechanisms for the observed beneficial effects of metformin application remain to be completely understood. The present study assessed whether metformin, a widely used therapeutic drug for T2DM, may accelerate wound closure in T2DM db/db mice. Genetically hyperglycemic db/db mice were used as the T2DM model. Metformin (250 mg/kg/day; intragastric) was administered for two weeks prior to EPC collection and wound model creation in db/db mice. Wound healing was evaluated by alterations in the wound area and the number of platelet endothelial cell adhesion molecule‑positive cells. The function of the isolated bone marrow‑derived EPCs (BM‑EPCs) was assessed by a tube formation assay. The number of circulating EPCs, and the levels of intracellular nitric oxide (NO) and superoxide (O2‑) were detected by flow cytometry. Thrombospondin‑1 (TSP‑1) expression was determined by western blot analysis. It was observed that treatment with metformin accelerated wound healing, improved angiogenesis and increased the circulating EPC number in db/db mice. In vitro, treatment with metformin reversed the impaired BM‑EPC function reflected by tube formation, and significantly increased NO production while decreasing O2‑ levels in BM‑EPCs from db/db mice. In addition, TSP‑1 expression was markedly attenuated by treatment with metformin in cultured BM‑EPCs. Metformin contributed to wound healing and improved angiogenesis in T2DM mice, which was, in part, associated with stimulation of NO, and inhibition of O2‑ and TSP‑1 in EPCs from db/db mice.

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1	Chan JC, Cho NH, Tajima N and Shaw J: Diabetes in the Western Pacific Region-past, present and future. Diabetes Res Clin Pract. 103:244–255. 2014. View Article : Google Scholar : PubMed/NCBI
2	Thandavarayan RA, Garikipati VN, Joladarashi D, Babu S Suresh, Jeyabal P, Verma SK, Mackie AR, Khan M, Arumugam S, Watanabe K, et al: Sirtuin-6 deficiency exacerbates diabetes-induced impairment of wound healing. Exp Dermatol. 24:773–778. 2015. View Article : Google Scholar : PubMed/NCBI
3	Papanas N, Demetzos C, Pippa N, Maltezos E and Tentolouris N: Efficacy of a new heparan sulfate mimetic dressing in the healing of foot and lower extremity ulcerations in type 2 diabetes: A case series. Int J Low Extrem Wounds. 15:63–67. 2016. View Article : Google Scholar : PubMed/NCBI
4	Zgheib C and Liechty KW: Shedding light on miR-26a: Another key regulator of angiogenesis in diabetic wound healing. J Mol Cell Cardiol. 92:203–205. 2016. View Article : Google Scholar : PubMed/NCBI
5	Li DW, Liu ZQ, Wei J, Liu Y and Hu LS: Contribution of endothelial progenitor cells to neovascularization (Review). Int J Mol Med. 30:1000–1006. 2012. View Article : Google Scholar : PubMed/NCBI
6	Fadini GP, Sartore S, Albiero M, Baesso I, Murphy E, Menegolo M, Grego F, de Kreutzenberg S Vigili, Tiengo A, Agostini C and Avogaro A: Number and function of endothelial progenitor cells as a marker of severity for diabetic vasculopathy. Arterioscler Thromb Vasc Biol. 26:2140–2146. 2006. View Article : Google Scholar : PubMed/NCBI
7	Kovacic JC, Moore J, Herbert A, Ma D, Boehm M and Graham RM: Endothelial progenitor cells, angioblasts, and angiogenesis-old terms reconsidered from a current perspective. Trends Cardiovasc Med. 18:45–51. 2008. View Article : Google Scholar : PubMed/NCBI
8	Liao YF, Chen LL, Zeng TS, Li YM, Fan Yu, Hu LJ and Ling Yue: Number of circulating endothelial progenitor cells as a marker of vascular endothelial function for type 2 diabetes. Vasc Med. 15:279–285. 2010. View Article : Google Scholar : PubMed/NCBI
9	Tepper OM: Human endothelial progenitor cells from type II diabetics exhibit impaired proliferation, adhesion, and incorporation into vascular structures. Circulation. 106:2781–2786. 2002. View Article : Google Scholar : PubMed/NCBI
10	Wang CH, Ting MK, Verma S, Kuo LT, Yang NI, Hsieh IC, Wang SY, Hung A and Cherng WJ: Pioglitazone increases the numbers and improves the functional capacity of endothelial progenitor cells in patients with diabetes mellitus. Am Heart J. 152:1051.e1–8. 2006. View Article : Google Scholar
11	Fadini GP, Miorin M, Facco M, Bonamico S, Baesso I, Grego F, Menegolo M, de Kreutzenberg SV, Tiengo A, Agostini C and Avogaro A: Circulating endothelial progenitor cells are reduced in peripheral vascular complications of type 2 diabetes mellitus. J Am Coll Cardiol. 45:1449–1457. 2005. View Article : Google Scholar : PubMed/NCBI
12	Nakamura K, Oe H, Kihara H, Shimada K, Fukuda S, Watanabe K, Takagi T, Yunoki K, Miyoshi T, Hirata K, et al: DPP-4 inhibitor and alpha-glucosidase inhibitor equally improve endothelial function in patients with type 2 diabetes: EDGE study. Cardiovasc Diabetol. 13:1102014. View Article : Google Scholar : PubMed/NCBI
13	Yue WS, Lau KK, Siu CW, Wang M, Yan GH, Yiu KH and Tse HF: Impact of glycemic control on circulating endothelial progenitor cells and arterial stiffness in patients with type 2 diabetes mellitus. Cardiovasc Diabetol. 10:1132011. View Article : Google Scholar : PubMed/NCBI
14	Badr G, Hozzein WN, Badr BM, Al Ghamdi A, Eldien HM Saad and Garraud O: Bee venom accelerates wound healing in diabetic mice by suppressing activating transcription factor-3 (ATF-3) and inducible nitric oxide synthase (iNOS)-mediated oxidative stress and recruiting bone marrow-derived endothelial progenitor cells. J Cell Physiol. 231:2159–2171. 2016. View Article : Google Scholar : PubMed/NCBI
15	Gallagher KA, Goldstein LJ, Thom SR and Velazquez OC: Hyperbaric oxygen and bone marrow-derived endothelial progenitor cells in diabetic wound healing. Vascular. 14:328–337. 2006. View Article : Google Scholar : PubMed/NCBI
16	Gallagher KA, Liu ZJ, Xiao M, Chen H, Goldstein LJ, Buerk DG, Nedeau A, Thom SR and Velazquez OC: Diabetic impairments in NO-mediated endothelial progenitor cell mobilization and homing are reversed by hyperoxia and SDF-1 alpha. J Clin Invest. 117:1249–1259. 2007. View Article : Google Scholar : PubMed/NCBI
17	Hattori Y, Hattori K and Hayashi T: Pleiotropic benefits of metformin: Macrophage targeting its anti-inflammatory mechanisms. Diabetes. 64:1907–1909. 2015. View Article : Google Scholar : PubMed/NCBI
18	Li DJ, Huang F, Lu WJ, Jiang GJ, Deng YP and Shen FM: Metformin promotes irisin release from murine skeletal muscle independently of AMP-activated protein kinase activation. Acta Physiol (Oxf). 213:711–721. 2015. View Article : Google Scholar : PubMed/NCBI
19	Chen LL, Liao YF, Zeng TS, Yu F, Li HQ and Feng Y: Effects of metformin plus gliclazide compared with metformin alone on circulating endothelial progenitor cell in type 2 diabetic patients. Endocrine. 38:266–275. 2010. View Article : Google Scholar : PubMed/NCBI
20	Varma V, Yao-Borengasser A, Bodles AM, Rasouli N, Phanavanh B, Nolen GT, Kern EM, Nagarajan R, Spencer HJ III, Lee MJ, et al: Thrombospondin-1 is an adipokine associated with obesity, adipose inflammation, and insulin resistance. Diabetes. 57:432–439. 2008. View Article : Google Scholar : PubMed/NCBI
21	Dabir P, Marinic TE, Krukovets I and Stenina OI: Aryl hydrocarbon receptor is activated by glucose and regulates the thrombospondin-1 gene promoter in endothelial cells. Circ Res. 102:1558–1565. 2008. View Article : Google Scholar : PubMed/NCBI
22	Stenina OI, Krukovets I, Wang K, Zhou Z, Forudi F, Penn MS, Topol EJ and Plow EF: Increased expression of thrombospondin-1 in vessel wall of diabetic Zucker rat. Circulation. 107:3209–3215. 2003. View Article : Google Scholar : PubMed/NCBI
23	Tie L, Chen LY, Chen DD, Xie HH, Channon KM and Chen AF: GTP cyclohydrolase I prevents diabetic-impaired endothelial progenitor cells and wound healing by suppressing oxidative stress/thrombospondin-1. Am J Physiol Endocrinol Metab. 306:E1120–E1131. 2014. View Article : Google Scholar : PubMed/NCBI
24	Tan BK, Adya R, Chen J, Farhatullah S, Heutling D, Mitchell D, Lehnert H and Randeva HS: Metformin decreases angiogenesis via NF-kappaB and Erk1/2/Erk5 pathways by increasing the antiangiogenic thrombospondin-1. Cardiovasc Res. 83:566–574. 2009. View Article : Google Scholar : PubMed/NCBI
25	Xie HH, Zhou S, Chen DD, Channon KM, Su DF and Chen AF: GTP cyclohydrolase I/BH4 pathway protects EPCs via suppressing oxidative stress and thrombospondin-1 in salt-sensitive hypertension. Hypertension. 56:1137–1144. 2010. View Article : Google Scholar : PubMed/NCBI
26	Li ZP, Xin RJ, Yang H, Jiang GJ, Deng YP, Li DJ and Shen FM: Diazoxide accelerates wound healing by improving EPC function. Front Biosci (Landmark Ed). 21:1039–1051. 2016. View Article : Google Scholar : PubMed/NCBI
27	Cai J, Lu S, Yao Z, Deng YP, Zhang LD, Yu JW, Ren GF, Shen FM and Jiang GJ: Glibenclamide attenuates myocardial injury by lipopolysaccharides in streptozotocin-induced diabetic mice. Cardiovasc Diabetol. 13:1062014. View Article : Google Scholar : PubMed/NCBI
28	Marrotte EJ, Chen DD, Hakim JS and Chen AF: Manganese superoxide dismutase expression in endothelial progenitor cells accelerates wound healing in diabetic mice. J Clin Invest. 120:4207–4219. 2010. View Article : Google Scholar : PubMed/NCBI
29	Chen JK, Deng YP, Jiang GJ, Liu YZ, Zhao T and Shen FM: Establishment of tube formation assay of bone marrow-derived endothelial progenitor cells. CNS Neurosci Ther. 19:533–535. 2013. View Article : Google Scholar : PubMed/NCBI
30	Lee MH, Choi EN, Jeon YJ and Jung SC: Possible role of transforming growth factor-β1 and vascular endothelial growth factor in Fabry disease nephropathy. Int J Mol Med. 30:1275–1280. 2012. View Article : Google Scholar : PubMed/NCBI
31	Aoyama H, Daitoku H and Fukamizu A: Nutrient control of phosphorylation and translocation of Foxo1 in C57BL/6 and db/db mice. Int J Mol Med. 18:433–439. 2006.PubMed/NCBI
32	Bao Q, Shen X, Qian L, Gong C, Nie M and Dong Y: Anti-diabetic activities of catalpol in db/db mice. Korean J Physiol Pharmacol. 20:153–160. 2016. View Article : Google Scholar : PubMed/NCBI
33	Tamura Y, Murayama T, Minami M, Yokode M and Arai H: Differential effect of statins on diabetic nephropathy in db/db mice. Int J Mol Med. 28:683–687. 2011.PubMed/NCBI
34	Galiano RD, Tepper OM, Pelo CR, Bhatt KA, Callaghan M, Bastidas N, Bunting S, Steinmetz HG and Gurtner GC: Topical vascular endothelial growth factor accelerates diabetic wound healing through increased angiogenesis and by mobilizing and recruiting bone marrow-derived cells. Am J Pathol. 164:1935–1947. 2004. View Article : Google Scholar : PubMed/NCBI
35	Galeano M, Altavilla D, Cucinotta D, Russo GT, Calò M, Bitto A, Marini H, Marini R, Adamo EB, Seminara P, et al: Recombinant human erythropoietin stimulates angiogenesis and wound healing in the genetically diabetic mouse. Diabetes. 53:2509–2517. 2004. View Article : Google Scholar : PubMed/NCBI
36	Zhang XN, Ma ZJ, Wang Y, Li YZ, Sun B, Guo X, Pan CQ and Chen LM: The four-herb Chinese medicine formula Tuo-Li-Xiao-Du-San accelerates cutaneous wound healing in streptozotocin-induced diabetic rats through reducing inflammation and increasing angiogenesis. J Diabetes Res. 2016:56391292016. View Article : Google Scholar : PubMed/NCBI
37	Caligiuri G, Groyer E, Khallou-Laschet J, Al Haj Zen A, Sainz J, Urbain D, Gaston AT, Lemitre M, Nicoletti A and Lafont A: Reduced immunoregulatory CD31+ T cells in the blood of atherosclerotic mice with plaque thrombosis. Arterioscler Thromb Vasc Biol. 25:1659–1664. 2005. View Article : Google Scholar : PubMed/NCBI
38	Tellechea A, Kafanas A, Leal EC, Tecilazich F, Kuchibhotla S, Auster ME, Kontoes I, Paolino J, Carvalho E, Nabzdyk LP and Veves A: Increased skin inflammation and blood vessel density in human and experimental diabetes. Int J Low Extrem Wounds. 12:4–11. 2013. View Article : Google Scholar : PubMed/NCBI
39	Desouza CV: Does drug therapy reverse endothelial progenitor cell dysfunction in diabetes? J Diabetes Complications. 27:519–525. 2013. View Article : Google Scholar : PubMed/NCBI
40	Lin JT, Chen HM, Chiu CH and Liang YJ: AMP-activated protein kinase activators in diabetic ulcers: From animal studies to Phase II drugs under investigation. Expert Opin Investig Drugs. 23:1253–1265. 2014. View Article : Google Scholar : PubMed/NCBI
41	Ochoa-Gonzalez F, Cervantes-Villagrana AR, Fernandez-Ruiz JC, Nava-Ramirez HS, Hernandez-Correa AC, Enciso-Moreno JA and Castañeda-Delgado JE: Metformin induces cell cycle arrest, reduced proliferation, wound healing impairment in vivo and is associated to clinical outcomes in diabetic foot ulcer patients. PLoS One. 11:e01509002016. View Article : Google Scholar : PubMed/NCBI
42	Mao L, Huang M, Chen SC, Li YN, Xia YP, He QW, Wang MD, Huang Y, Zheng L and Hu B: Endogenous endothelial progenitor cells participate in neovascularization via CXCR4/SDF-1 axis and improve outcome after stroke. CNS Neurosci Ther. 20:460–468. 2014. View Article : Google Scholar : PubMed/NCBI
43	Kim KA, Shin YJ, Kim JH, Lee H, Noh SY, Jang SH and Bae ON: Dysfunction of endothelial progenitor cells under diabetic conditions and its underlying mechanisms. Arch Pharm Res. 35:223–234. 2012. View Article : Google Scholar : PubMed/NCBI
44	Ghosh S, Lakshmanan AP, Hwang MJ, Kubba H, Mushannen A, Triggle CR and Ding H: Metformin improves endothelial function in aortic tissue and microvascular endothelial cells subjected to diabetic hyperglycaemic conditions. Biochem Pharmacol. 98:412–421. 2015. View Article : Google Scholar : PubMed/NCBI
45	Shi Y, He Z, Jia Z and Xu C: Inhibitory effect of metformin combined with gemcitabine on pancreatic cancer cells in vitro and in vivo. Mol Med Rep. 14:2921–2928. 2016. View Article : Google Scholar : PubMed/NCBI
46	Gao L, Li P, Zhang J, Hagiwara M, Shen B, Bledsoe G, Chang E, Chao L and Chao J: Novel role of kallistatin in vascular repair by promoting mobility, viability, and function of endothelial progenitor cells. J Am Heart Assoc. 3:e0011942014. View Article : Google Scholar : PubMed/NCBI
47	Bhattacharyya S, Marinic TE, Krukovets I, Hoppe G and Stenina OI: Cell type-specific post-transcriptional regulation of production of the potent antiangiogenic and proatherogenic protein thrombospondin-1 by high glucose. J Biol Chem. 283:5699–5707. 2008. View Article : Google Scholar : PubMed/NCBI
48	Isenberg JS, Wink DA and Roberts DD: Thrombospondin-1 antagonizes nitric oxide-stimulated vascular smooth muscle cell responses. Cardiovasc Res. 71:785–793. 2006. View Article : Google Scholar : PubMed/NCBI
49	Ridnour LA, Isenberg JS, Espey MG, Thomas DD, Roberts DD and Wink DA: Nitric oxide regulates angiogenesis through a functional switch involving thrombospondin-1. Proc Natl Acad Sci USA. 102:pp. 13147–13152. 2005; View Article : Google Scholar : PubMed/NCBI
50	Xu L, Xun G, Yao Z, Liu Y, Qiu Y, Liu K, Zhu D, Gu Q, Xu X and Ho PC: Effects of generated trans-arachidonic acids on retinal capillary during nitrative stress in diabetic rats. Ophthalmologica. 222:37–41. 2008. View Article : Google Scholar : PubMed/NCBI