FRINK Linked Data Fragments server

Matches in SemOpenAlex for { <https://semopenalex.org/work/W2955275274> ?p ?o ?g. }

• W2955275274 endingPage "234.e7" @default.
• W2955275274 startingPage "223" @default.
• W2955275274 abstract "Skin ulcers resulting from impaired wound healing are a serious complication of diabetes. Unresolved inflammation, associated with the dysregulation of both the phenotype and function of macrophages, is involved in the poor healing of diabetic wounds. Here, we report that topical pharmacological inhibition of the mineralocorticoid receptor (MR) by canrenoate or MR small interfering RNA can resolve inflammation to improve delayed skin wound healing in diabetic mouse models; importantly, wounds from normal mice are unaffected. The beneficial effect of canrenoate is associated with an increased ratio of anti-inflammatory M2 macrophages to proinflammatory M1 macrophages in diabetic wounds. Furthermore, we show that MR blockade leads to downregulation of the MR target, LCN2, which may facilitate macrophage polarization toward the M2 phenotype and improve impaired angiogenesis in diabetic wounds. Indeed, diabetic LCN2-deficient mice showed improved wound healing associated with macrophage M2 polarization and angiogenesis. In addition, recombinant LCN2 protein prevented IL-4–induced macrophage switch from M1 to M2 phenotype. In conclusion, topical MR blockade accelerates skin wound healing in diabetic mice via LCN2 reduction, M2 macrophage polarization, prevention of inflammation, and induction of angiogenesis. Skin ulcers resulting from impaired wound healing are a serious complication of diabetes. Unresolved inflammation, associated with the dysregulation of both the phenotype and function of macrophages, is involved in the poor healing of diabetic wounds. Here, we report that topical pharmacological inhibition of the mineralocorticoid receptor (MR) by canrenoate or MR small interfering RNA can resolve inflammation to improve delayed skin wound healing in diabetic mouse models; importantly, wounds from normal mice are unaffected. The beneficial effect of canrenoate is associated with an increased ratio of anti-inflammatory M2 macrophages to proinflammatory M1 macrophages in diabetic wounds. Furthermore, we show that MR blockade leads to downregulation of the MR target, LCN2, which may facilitate macrophage polarization toward the M2 phenotype and improve impaired angiogenesis in diabetic wounds. Indeed, diabetic LCN2-deficient mice showed improved wound healing associated with macrophage M2 polarization and angiogenesis. In addition, recombinant LCN2 protein prevented IL-4–induced macrophage switch from M1 to M2 phenotype. In conclusion, topical MR blockade accelerates skin wound healing in diabetic mice via LCN2 reduction, M2 macrophage polarization, prevention of inflammation, and induction of angiogenesis. Impaired cutaneous wound healing, responsible for chronic ulcers, represents one of the most important complications of diabetes. Indeed, the incidence of diabetic foot ulcers is increasing because of the high prevalence of diabetes mellitus worldwide and the longer life expectancy of patients (Boulton et al., 2005Boulton A.J. Vileikyte L. Ragnarson-Tennvall G. Apelqvist J. The global burden of diabetic foot disease.Lancet. 2005; 366: 1719-1724Abstract Full Text Full Text PDF PubMed Scopus (1474) Google Scholar). The prevalence of global diabetic foot ulcers was recently reported to be 6.3% and up to 13.0% in North America (Zhang et al., 2017Zhang P. Lu J. Jing Y. Tang S. Zhu D. Bi Y. Global epidemiology of diabetic foot ulceration: a systematic review and meta-analysis.Ann Med. 2017; 49: 106-116Crossref PubMed Scopus (438) Google Scholar). Ulcers remain in a chronic inflammation state that prevents them from healing, resulting in other severe complications such as pain, infection, and eventually amputation (Greenhalgh, 2003Greenhalgh D.G. Wound healing and diabetes mellitus.Clin Plast Surg. 2003; 30: 37-45Abstract Full Text Full Text PDF PubMed Scopus (253) Google Scholar, Noor et al., 2017Noor S. Khan R.U. Ahmad J. Understanding diabetic foot infection and its management.Diabetes Metab Syndr. 2017; 11: 149-156Crossref PubMed Scopus (40) Google Scholar, Peters and Lipsky, 2013Peters E.J.G. Lipsky B.A. Diagnosis and management of infection in the diabetic foot.Med Clin North Am. 2013; 97: 911-946Abstract Full Text Full Text PDF PubMed Scopus (60) Google Scholar). Previous studies showed that most diabetic amputations are preceded by a foot ulceration, which subsequently results in serious gangrene or infection (Boulton et al., 2005Boulton A.J. Vileikyte L. Ragnarson-Tennvall G. Apelqvist J. The global burden of diabetic foot disease.Lancet. 2005; 366: 1719-1724Abstract Full Text Full Text PDF PubMed Scopus (1474) Google Scholar, Lepantalo et al., 2011Lepantalo M. Apelqvist J. Setacci C. Ricco J.B. de Donato G. Becker F. et al.Chapter V: Diabetic foot.Eur J Vasc Endovasc Surg. 2011; 42: S60-S74Abstract Full Text Full Text PDF PubMed Scopus (117) Google Scholar). Diabetic foot ulcers are thus a major health issue that significantly affects the lives of patients, resulting in a high financial burden in many countries (Boulton et al., 2005Boulton A.J. Vileikyte L. Ragnarson-Tennvall G. Apelqvist J. The global burden of diabetic foot disease.Lancet. 2005; 366: 1719-1724Abstract Full Text Full Text PDF PubMed Scopus (1474) Google Scholar). The prevention and appropriate care of diabetic ulcers are thus of major importance. Several therapies have been proposed for diabetic ulcers but effective treatment is still needed (Clokie et al., 2017Clokie M. Greenway A.L. Harding K. Jones N.J. Vedhara K. Game F. et al.New horizons in the understanding of the causes and management of diabetic foot disease: report from the 2017 Diabetes UK Annual Professional Conference Symposium.Diabet Med. 2017; 34: 305-315Crossref PubMed Scopus (15) Google Scholar, Dinh et al., 2012Dinh T. Tecilazich F. Kafanas A. Doupis J. Gnardellis C. Leal E. et al.Mechanisms involved in the development and healing of diabetic foot ulceration.Diabetes. 2012; 61: 2937-2947Crossref PubMed Scopus (204) Google Scholar, Galiano et al., 2004Galiano R.D. Tepper O.M. Pelo C.R. Bhatt K.A. Callaghan M. Bastidas N. et al.Topical vascular endothelial growth factor accelerates diabetic wound healing through increased angiogenesis and by mobilizing and recruiting bone marrow-derived cells.Am J Pathol. 2004; 164: 1935-1947Abstract Full Text Full Text PDF PubMed Scopus (580) Google Scholar). The lack of therapeutic tools likely results from the complex mechanisms involved in the development of unhealed wounds. Many pathogenic factors, such as vascular defects and neuropathy, are responsible for diabetic ulcers (Ahmed and Antonsen, 2016Ahmed A.S. Antonsen E.L. Immune and vascular dysfunction in diabetic wound healing.J Wound Care. 2016; 25: S35-S46Crossref PubMed Google Scholar, Boulton, 2014Boulton A.J.M. Diabetic neuropathy and foot complications.Handb Clin Neurol. 2014; 126: 97-107Crossref PubMed Scopus (46) Google Scholar, Brem and Tomic-Canic, 2007Brem H. Tomic-Canic M. Cellular and molecular basis of wound healing in diabetes.J Clin Invest. 2007; 117: 1219-1222Crossref PubMed Scopus (987) Google Scholar, Dinh et al., 2012Dinh T. Tecilazich F. Kafanas A. Doupis J. Gnardellis C. Leal E. et al.Mechanisms involved in the development and healing of diabetic foot ulceration.Diabetes. 2012; 61: 2937-2947Crossref PubMed Scopus (204) Google Scholar). A chronic inflammatory environment is also a common feature observed in unhealed wounds and is mainly associated with the uncontrolled recruitment and activation of inflammatory cells, in particular monocytes and macrophages (Boniakowski et al., 2017Boniakowski A.E. Kimball A.S. Jacobs B.N. Kunkel S.L. Gallagher K.A. Macrophage-mediated inflammation in normal and diabetic wound healing.J Immunol. 2017; 199: 17-24Crossref PubMed Scopus (143) Google Scholar, Leal et al., 2015Leal E.C. Carvalho E. Tellechea A. Kafanas A. Tecilazich F. Kearney C. et al.Substance P promotes wound healing in diabetes by modulating inflammation and macrophage phenotype.Am J Pathol. 2015; 185: 1638-1648Abstract Full Text Full Text PDF PubMed Scopus (103) Google Scholar, Okizaki et al., 2015Okizaki S. Ito Y. Hosono K. Oba K. Ohkubo H. Amano H. et al.Suppressed recruitment of alternatively activated macrophages reduces TGF-beta1 and impairs wound healing in streptozotocin-induced diabetic mice.Biomed Pharmacother. 2015; 70: 317-325Crossref PubMed Scopus (68) Google Scholar). During the wound repair process, macrophages present various phenotypes and functions, depending on the stage of the healing response and how they are activated. During the early phase of wound healing, classically activated macrophages, known as M1 macrophages, are recruited and secrete proinflammatory cytokines to kill pathogens and clear away the damaged tissue. In contrast, alternatively activated M2 macrophages secrete anti-inflammatory factors to resolve inflammation and produce factors required for later regenerative phases. The balance between these macrophage subpopulations is pivotal for maintaining a physiological healing process (Gordon, 2003Gordon S. Alternative activation of macrophages.Nat Rev Immunol. 2003; 3: 23-35Crossref PubMed Scopus (4482) Google Scholar, Mahdavian Delavary et al., 2011Mahdavian Delavary B. van der Veer W.M. van Egmond M. Niessen F.B. Beelen R.H.J. Macrophages in skin injury and repair.Immunobiology. 2011; 216: 753-762Crossref PubMed Scopus (446) Google Scholar, Mantovani et al., 2004Mantovani A. Sica A. Sozzani S. Allavena P. Vecchi A. Locati M. The chemokine system in diverse forms of macrophage activation and polarization.Trends Immunol. 2004; 25: 677-686Abstract Full Text Full Text PDF PubMed Scopus (4071) Google Scholar). However, in unhealed wounds, such as diabetic ulcers, macrophages are chronically activated and restrained to the M1 phenotype, heavily contributing to the chronic inflammatory microenvironment observed in these wounds. Moreover, such prolonged inflammation delays the process of tissue regeneration, including re-epithelialization, granulation tissue formation, and vascularization (Boniakowski et al., 2017Boniakowski A.E. Kimball A.S. Jacobs B.N. Kunkel S.L. Gallagher K.A. Macrophage-mediated inflammation in normal and diabetic wound healing.J Immunol. 2017; 199: 17-24Crossref PubMed Scopus (143) Google Scholar, He et al., 2017He R. Yin H. Yuan B. Liu T. Luo L. Huang P. et al.IL-33 improves wound healing through enhanced M2 macrophage polarization in diabetic mice.Mol Immunol. 2017; 90: 42-49Crossref PubMed Scopus (54) Google Scholar, Maruyama et al., 2007Maruyama K. Asai J. Ii M. Thorne T. Losordo D.W. D'Amore P.A. Decreased macrophage number and activation lead to reduced lymphatic vessel formation and contribute to impaired diabetic wound healing.Am J Pathol. 2007; 170: 1178-1191Abstract Full Text Full Text PDF PubMed Scopus (342) Google Scholar, Okizaki et al., 2015Okizaki S. Ito Y. Hosono K. Oba K. Ohkubo H. Amano H. et al.Suppressed recruitment of alternatively activated macrophages reduces TGF-beta1 and impairs wound healing in streptozotocin-induced diabetic mice.Biomed Pharmacother. 2015; 70: 317-325Crossref PubMed Scopus (68) Google Scholar). Enhancing macrophage polarization toward the M2 phenotype may help to promote cellular proliferation and angiogenesis and to accelerate diabetic wound closure (He et al., 2017He R. Yin H. Yuan B. Liu T. Luo L. Huang P. et al.IL-33 improves wound healing through enhanced M2 macrophage polarization in diabetic mice.Mol Immunol. 2017; 90: 42-49Crossref PubMed Scopus (54) Google Scholar, Leal et al., 2015Leal E.C. Carvalho E. Tellechea A. Kafanas A. Tecilazich F. Kearney C. et al.Substance P promotes wound healing in diabetes by modulating inflammation and macrophage phenotype.Am J Pathol. 2015; 185: 1638-1648Abstract Full Text Full Text PDF PubMed Scopus (103) Google Scholar, Okizaki et al., 2015Okizaki S. Ito Y. Hosono K. Oba K. Ohkubo H. Amano H. et al.Suppressed recruitment of alternatively activated macrophages reduces TGF-beta1 and impairs wound healing in streptozotocin-induced diabetic mice.Biomed Pharmacother. 2015; 70: 317-325Crossref PubMed Scopus (68) Google Scholar). The mineralocorticoid receptor (MR), a steroid receptor that belongs to the nuclear receptor superfamily of transcription factors, plays a key role in various physiological and pathophysiological phenomena, including obesity- and diabetes-related complications, such as vascular dysfunction, insulin resistance, metabolic disorders, and chronic inflammation (Guo et al., 2008Guo C. Ricchiuti V. Lian B.Q. Yao T.M. Coutinho P. Romero J.R. et al.Mineralocorticoid receptor blockade reverses obesity-related changes in expression of adiponectin, peroxisome proliferator-activated receptor-γ, and proinflammatory adipokines.Circulation. 2008; 117: 2253-2261Crossref PubMed Scopus (244) Google Scholar, Hirata et al., 2009Hirata A. Maeda N. Hiuge A. Hibuse T. Fujita K. Okada T. et al.Blockade of mineralocorticoid receptor reverses adipocyte dysfunction and insulin resistance in obese mice.Cardiovasc Res. 2009; 84: 164-172Crossref PubMed Scopus (162) Google Scholar, Jaisser and Farman, 2015Jaisser F. Farman N. Emerging roles of the mineralocorticoid receptor in pathology: toward new paradigms in Clinical Pharmacology.Pharmacol Rev. 2015; 68: 49-75Crossref Scopus (138) Google Scholar). We recently reported the involvement of MR activation in delayed wound closure in type 1 diabetes and the improvement of impaired wound re-epithelialization following local application of an MR antagonist to the wounds (Nguyen et al., 2016Nguyen V.T. Farman N. Maubec E. Nassar D. Desposito D. Waeckel L. et al.Re-epithelialization of pathological cutaneous wounds is improved by local mineralocorticoid receptor antagonism.J Invest Dermatol. 2016; 136: 2080-2089Abstract Full Text Full Text PDF PubMed Scopus (24) Google Scholar). However, we did not explore the integrated impact on either the epidermis or dermis in diabetic wounds or the underlying mechanisms of the benefit of local MR antagonism. It is not known whether MR acts on the inflammatory response and dermal angiogenesis during wound repair in diabetes. Here, we analyzed the positive impact of topical pharmacological MR blockade on impaired wound healing of streptozotocin (STZ)-treated mice and of db-db mice, a model of type 2 diabetes. We focused on inflammation and angiogenesis and identified LCN2, a primary MR target (Buonafine et al., 2018Buonafine M. Martinez-Martinez E. Jaisser F. More than a simple biomarker: the role of NGAL in cardiovascular and renal diseases.Clin Sci. 2018; 132: 909-923Crossref PubMed Scopus (42) Google Scholar, Martínez-Martínez et al., 2017Martínez-Martínez E. Buonafine M. Boukhalfa I. Ibarrola J. Fernández-Celis A. Kolkhof P. et al.Aldosterone target NGAL (neutrophil gelatinase–associated lipocalin) is involved in cardiac remodeling after myocardial infarction through NFκB pathway.Hypertension. 2017; 70: 1148-1156Crossref PubMed Scopus (39) Google Scholar), as a potential mechanism involved in these beneficial effects. We have shown previously that local MR blockade with canrenoate improves the delayed wound re-epithelialization of type 1 diabetic mice (STZ mice) (Nguyen et al., 2016Nguyen V.T. Farman N. Maubec E. Nassar D. Desposito D. Waeckel L. et al.Re-epithelialization of pathological cutaneous wounds is improved by local mineralocorticoid receptor antagonism.J Invest Dermatol. 2016; 136: 2080-2089Abstract Full Text Full Text PDF PubMed Scopus (24) Google Scholar). To exclude MR-independent effects of canrenoate, we designed an in vivo experimental strategy to interfere specifically with the MR. Topical treatment with MR small interfering RNA (siRNA) blocked MR upregulation in STZ wounds (Supplementary Figure S1a) and restored the defective re-epithelialization of STZ mice, as compared with those of diabetic mice treated by scrambled siRNA (Supplementary Figure S1c–g). These results are quite comparable to those previously obtained using canrenoate, indicating that MR antagonism and silencing is beneficial for wounding in type 1 diabetic mice. Next, we questioned the effect of canrenoate on the delayed wound healing of type 2 diabetes using db-db mice, a genetic mouse model of type 2 diabetes. MR mRNA expression was higher in wounded skin and the surrounding normal skin of db-db mice than that of normal db/+ control (CT) mice (Figure 1a), raising the question of the role of MR overexpression in abnormal wound repair in type 2 diabetes. Wound healing was strongly impaired in db-db mice relative to that of CT mice (Figure 1b and c). Consistent with our observations in STZ mice, local treatment with canrenoate improved the wound healing delay in db-db mice, whereas it did not modify wound closure in normal CT mice (Figure 1b and c). Moreover, keratin-14 staining showed that impaired re-epithelialization of db-db wounds was rescued by local canrenoate treatment, as illustrated by the longer neoepidermis at the edges of the wound sections (Figure 1d and e) and the shorter residual wound length (the distance between two edges) in canrenoate-treated db-db mice than in phosphate buffered saline–treated db-db mice (Figure 1d and f). This reduction in wound surface is indicative of re-epithelialization rather than wound contraction. The improvement of re-epithelialization was accompanied by a higher number of proliferating Ki67-positive keratinocytes in the neoepidermis of the wounds of canrenoate-treated than phosphate buffered saline–treated db-db mice (Figure 1g and h). These observations suggest that the effects of topical canrenoate treatment on diabetic wound healing are mediated by MR signaling. Thus, topical MR blockade restored the impaired proliferation of epidermal keratinocytes and blunted the delayed re-epithelialization of wounds in 2 diabetes mouse models. In contrast, these phenomena were not found in wounds of normal CT mice. Impaired dermal wound angiogenesis is a key contributor to the wound healing defect of diabetes. Herein, we explored the role of MR in the defective wound angiogenesis of diabetic mouse models. The density of CD31+ blood vessels was lower in the wounds of STZ-induced (Figure 2a in red, 2b) and db-db diabetic mice than those of CT mice (Figure 2e and f), consistent with previous reports. Decreased vessel density of diabetic wounds was partly rescued by topical treatment of these wounds with canrenoate (Figure 2a, b, e, and f). MR inactivation by local MR siRNA treatment of STZ wounds also showed a beneficial effect on wound angiogenesis (Supplementary Figure S1h and i). The improvement of vessel density in canrenoate-treated diabetic wounds was associated with an increased number of CD45-CD31+ endothelial cells in the wound beds, as quantified by FACS analysis of wound specimens (Figure 2c, d, g, and h). Moreover, by quantification of CD31+Ki67+ double positive cells on wound sections of STZ mice (Figure 2a, red and green), we found fewer proliferating endothelial cells in STZ wounds than in controls (2.87% ± 1.08, n = 7 vs 7.18% ± 0.71, n = 7, respectively), and this decrease was rescued by canrenoate treatment (STZ + phosphate buffered saline: 2.87% ± 1.08, n = 7 vs STZ + canrenoate: 6.37% ± 1.2, n = 7). These observations suggest that the activation of MR in the skin of diabetic mice is involved in the impaired wound angiogenesis of these animals, and that MR blockade could prevent such defects through increased proliferation of endothelial cells forming novel microvessels in wounds. Chronic inflammation is a common feature of diabetic ulcers. We examined whether MR plays a role in maintaining inflammation in diabetic wounds by studying the impact of MR blockade on the expression of some pro- and anti-inflammatory markers in wounds (Ashcroft et al., 2012Ashcroft G.S. Jeong M.J. Ashworth J.J. Hardman M. Jin W. Moutsopoulos N. et al.Tumor necrosis factor-alpha (TNF-α) is a therapeutic target for impaired cutaneous wound healing.Wound Repair Regen. 2012; 20: 38-49Crossref PubMed Scopus (139) Google Scholar, Barrientos et al., 2008Barrientos S. Stojadinovic O. Golinko M.S. Brem H. Tomic-Canic M. PERSPECTIVE ARTICLE: Growth factors and cytokines in wound healing.Wound Repair Regen. 2008; 16: 585-601Crossref PubMed Scopus (2138) Google Scholar, Ramalho et al., 2018Ramalho T. Filgueiras L. Silva-Jr I.A. Pessoa A.F.M. Jancar S. Impaired wound healing in type 1 diabetes is dependent on 5-lipoxygenase products.Sci Rep. 2018; 8: 14164Crossref PubMed Scopus (19) Google Scholar). Local treatment with canrenoate attenuated the overexpression of some proinflammatory genes, such as Tnfa, Mcpt1, Il12, and Il23, in diabetic wounds (Figure 3a and c). Consistent with this effect, the repression of anti-inflammatory factors in diabetic wounds was rescued by canrenoate treatment (Figure 3b and d). Thus, MR blockade blunted the inflammation observed in diabetic wounds and resulted in an anti-inflammatory status that could improve the impaired cellular proliferation and poor angiogenesis. A failure to switch from activated M1 macrophages to alternative M2 macrophages leads to chronic inflammation and impaired angiogenesis in various situations of delayed wound healing, including venous leg ulcers and diabetic foot ulcers (Guo et al., 2016Guo Y. Lin C. Xu P. Wu S. Fu X. Xia W. et al.AGEs induced autophagy impairs cutaneous wound healing via stimulating macrophage polarization to M1 in diabetes.Sci Rep. 2016; 6: 36416Crossref PubMed Scopus (55) Google Scholar, Okizaki et al., 2015Okizaki S. Ito Y. Hosono K. Oba K. Ohkubo H. Amano H. et al.Suppressed recruitment of alternatively activated macrophages reduces TGF-beta1 and impairs wound healing in streptozotocin-induced diabetic mice.Biomed Pharmacother. 2015; 70: 317-325Crossref PubMed Scopus (68) Google Scholar, Sindrilaru et al., 2011Sindrilaru A. Peters T. Wieschalka S. Baican C. Baican A. Peter H. et al.An unrestrained proinflammatory M1 macrophage population induced by iron impairs wound healing in humans and mice.J Clin Invest. 2011; 121: 985-997Crossref PubMed Scopus (652) Google Scholar). We investigated whether the beneficial effect of canrenoate treatment was associated with a switch of activated M1 macrophages toward the alternative M2 macrophage population. Wounds from both STZ and db-db mice showed more M1 macrophages (Ly6Chi) associated with fewer M2 macrophages (Ly6Clow) than CT mice, whereas the total number of macrophages in diabetic wounds was not different from that in control wounds (Figure 4). These observations are consistent with a number of previous reports showing the accumulation and persistence of activated M1 macrophages in diabetic wounds (Guo et al., 2016Guo Y. Lin C. Xu P. Wu S. Fu X. Xia W. et al.AGEs induced autophagy impairs cutaneous wound healing via stimulating macrophage polarization to M1 in diabetes.Sci Rep. 2016; 6: 36416Crossref PubMed Scopus (55) Google Scholar, Leal et al., 2015Leal E.C. Carvalho E. Tellechea A. Kafanas A. Tecilazich F. Kearney C. et al.Substance P promotes wound healing in diabetes by modulating inflammation and macrophage phenotype.Am J Pathol. 2015; 185: 1638-1648Abstract Full Text Full Text PDF PubMed Scopus (103) Google Scholar, Okizaki et al., 2015Okizaki S. Ito Y. Hosono K. Oba K. Ohkubo H. Amano H. et al.Suppressed recruitment of alternatively activated macrophages reduces TGF-beta1 and impairs wound healing in streptozotocin-induced diabetic mice.Biomed Pharmacother. 2015; 70: 317-325Crossref PubMed Scopus (68) Google Scholar). Importantly, the polarization of diabetic-wound macrophages was nearly restored to the levels of CT mice by topical canrenoate treatment. Canrenoate blunted the increase in M1 macrophages in STZ and db-db wounds (Figure 4a, c, e, and g) and the associated decrease in M2 macrophages (Figure 4a, d, e, and h). MR blockade also blunted overexpression of the proinflammatory marker Ly6C by diabetic-wound macrophages (Supplementary Figure S2a and b). These results suggest that topical MR blockade promotes a shift of proinflammatory M1 macrophages toward the anti-inflammatory M2 phenotype to resolve inflammation, promote angiogenesis, and accelerate wound healing. In addition to their pathogen-killing activity, macrophages in wounds also promote cellular proliferation and tissue regeneration, including re-epithelialization and dermal vascularization (Gordon, 2003Gordon S. Alternative activation of macrophages.Nat Rev Immunol. 2003; 3: 23-35Crossref PubMed Scopus (4482) Google Scholar, Lucas et al., 2010Lucas T. Waisman A. Ranjan R. Roes J. Krieg T. Müller W. et al.Differential roles of macrophages in diverse phases of skin repair.J Immunol. 2010; 184: 3964-3977Crossref PubMed Scopus (669) Google Scholar). We tested whether canrenoate treatment improves delayed diabetic wound angiogenesis through the modulation of macrophage polarization by assessing a panel of proangiogenic factors in wound tissue. Five days after wounding, gene expression of proangiogenic factors was impaired in STZ wounds with a significant decrease of Fgf2, Plgf, Tie2, and Angpt2 mRNA levels (Figure 5a). Importantly, topical canrenoate application restored the expression of these factors in diabetic wounds (Figure 5a). We then isolated macrophages (CD11b+F4.80+Ly6G-) from wounded skin by FACS to analyze the expression of proangiogenic genes. Vegfa, Fgf2, Plgf, and Tie2 mRNA levels were significantly lower in macrophages isolated from STZ wounds than those from control wounds, whereas topical application of canrenoate restored the inadequate expression of these angiogenic factors (Figure 5b). We recently showed that LCN2 is a primary target of aldosterone and mineralocorticoid receptor signaling in many organs, and that LCN2 plays a key role in the action of mineralocorticoids in the cardiovascular system (Buonafine et al., 2018Buonafine M. Martinez-Martinez E. Jaisser F. More than a simple biomarker: the role of NGAL in cardiovascular and renal diseases.Clin Sci. 2018; 132: 909-923Crossref PubMed Scopus (42) Google Scholar). LCN2 mRNA and protein expression were much higher in diabetic wounds than in CT nondiabetic wounds (Figure 6a and b; Supplementary Figure S3a and b). Local canrenoate or MR siRNA treatment lowered the upregulated expression of LCN2 in diabetic wounds to near the level found in control wounds, indicating that LCN2 is also an MR target in diabetic skin (Figure 6a and b; Supplementary Figure S1b; Supplementary Figure S3). We assessed whether LCN2 is involved in the impaired healing of diabetic wounds using a global Lcn2-knockout (KO) mouse model. Wound closure in diabetic STZ mice was significantly impaired relative to that of CT nondiabetic mice, but LCN2 inactivation prevented the delay in wound healing in Lcn2-KO diabetic mice (Figure 6c and d). Of note, LCN2 deficiency did not affect wound healing in normal nondiabetic mice (Figure 6c and d). FACS analysis of wound specimens demonstrated that LCN2 inactivation resulted in M1 to M2 macrophage polarization; the increase of proinflammatory M1 macrophages in CT diabetic wounds was blunted in LCN2-deficient diabetic mice (Figure 6e), whereas the decrease of M2 macrophages in diabetic wounds was prevented (Figure 6f). Importantly, LCN2 deficiency prevented the impaired angiogenesis associated with diabetes, with the presence of more endothelial cells in the wounds of diabetic Lcn2-KO mice than those of CT diabetic mice (Figure 6g). Moreover, expression of the proangiogenic genes Vegfa, Fgf2, Plgf, and Tie2 was higher in STZ Lcn2-KO macrophages than in STZ CT mice (Figure 6h). We hypothesized that the MR target LCN2 participates in the deleterious effect of MR activation in diabetic wound healing by tipping macrophage functional and phenotypic polarization from proangiogenic M2 macrophages toward proinflammatory M1. Macrophages isolated from the peritoneum of wild-type mice were first pretreated with lipopolysaccharide to induce an M1 phenotype and then with IL-4 to switch them toward an M2 phenotype (Supplementary Figure S4a). FACS analysis showed that treatment with recombinant LCN2 resulted in a higher percentage of Ly6Chi M1 macrophages (Supplementary Figure S4b) together with fewer Ly6Clow M2 macrophages (Supplementary Figure S4c), indicating that recombinant LCN2 inhibited the ability of lipopolysaccharide-pretreated macrophages to switch to the M2 phenotype in response to IL-4. This was associated with decreased expression of the angiogenic genes Vegfa and Plgf (Supplementary Figure S4d and e). Overall, these data show that MR blockade controls the phenotypic polarization of macrophages toward a repair M2 phenotype and promotes dermal angiogenesis through modulation of the expression and activity of LCN2, thereby improving the delayed wound healing in diabetes (Supplementary Figure S5). By using 2 mouse models of diabetes (STZ-induced type 1 and db-db type 2 diabetic mice), we demonstrate that inflammation and impaired healing of diabetic wounds are associated with the activation of MR signaling. Topical inhibition of this pathway provided a clear benefit to improve healing of these pathological wounds. This effect acts via inducing the polarization of macrophages toward the M2 phenotype, helping to resolve inflammation and rescue the angiogenesis defect of diabetic wounds. Moreover, we identified the MR target LCN2 as one of the underlying signaling pathways. In contrast, MR blockades do not modify wound closure from normal mice. We propose that inadequate MR occupancy by exogenous glucocorticoid (GC) or locally produced GC, as well as enhanced MR expression, may explain the benefit of MR blockade in a variety of pathological situations such as dermocorticoid treatments, UV irradiation, diabetic delayed wound healing, and perhaps psoriatic skin or GC-treated psoriasis (Hannen et al., 2017Hannen R. Udeh-Momoh C. Upton J. Wright M. Michael A. Gulati A. et al.Dysfunctional skin-de" @default.
• W2955275274 created "2019-07-12" @default.
• W2955275274 creator A5032128238 @default.
• W2955275274 creator A5036542646 @default.
• W2955275274 creator A5051434350 @default.
• W2955275274 creator A5061827333 @default.
• W2955275274 creator A5080087674 @default.
• W2955275274 creator A5080480919 @default.
• W2955275274 creator A5084151252 @default.
• W2955275274 date "2020-01-01" @default.
• W2955275274 modified "2023-10-18" @default.
• W2955275274 title "Cutaneous Wound Healing in Diabetic Mice Is Improved by Topical Mineralocorticoid Receptor Blockade" @default.
• W2955275274 cites W1590183001 @default.
• W2955275274 cites W1933223805 @default.
• W2955275274 cites W1949832432 @default.
• W2955275274 cites W1969806743 @default.
• W2955275274 cites W1970987370 @default.
• W2955275274 cites W1976141939 @default.
• W2955275274 cites W1989483643 @default.
• W2955275274 cites W1997751192 @default.
• W2955275274 cites W1999759666 @default.
• W2955275274 cites W1999935182 @default.
• W2955275274 cites W2003206402 @default.
• W2955275274 cites W2006960578 @default.
• W2955275274 cites W2008485826 @default.
• W2955275274 cites W2012614217 @default.
• W2955275274 cites W2017787550 @default.
• W2955275274 cites W2019583561 @default.
• W2955275274 cites W2029434160 @default.
• W2955275274 cites W2038112109 @default.
• W2955275274 cites W2038689279 @default.
• W2955275274 cites W204511859 @default.
• W2955275274 cites W2052241333 @default.
• W2955275274 cites W2054969539 @default.
• W2955275274 cites W2055496662 @default.
• W2955275274 cites W2068973625 @default.
• W2955275274 cites W2073155902 @default.
• W2955275274 cites W2093705034 @default.
• W2955275274 cites W2102445277 @default.
• W2955275274 cites W2105916525 @default.
• W2955275274 cites W2109605961 @default.
• W2955275274 cites W2118654016 @default.
• W2955275274 cites W2128049871 @default.
• W2955275274 cites W2131226296 @default.
• W2955275274 cites W2131465407 @default.
• W2955275274 cites W2133963705 @default.
• W2955275274 cites W2141870249 @default.
• W2955275274 cites W2142041241 @default.
• W2955275274 cites W2159745626 @default.
• W2955275274 cites W2160276873 @default.
• W2955275274 cites W2161927295 @default.
• W2955275274 cites W2163042571 @default.
• W2955275274 cites W2168537112 @default.
• W2955275274 cites W2206859595 @default.
• W2955275274 cites W2246046235 @default.
• W2955275274 cites W2288213997 @default.
• W2955275274 cites W2291574527 @default.
• W2955275274 cites W2293309775 @default.
• W2955275274 cites W2413944051 @default.
• W2955275274 cites W2434692906 @default.
• W2955275274 cites W2470441996 @default.
• W2955275274 cites W2501777749 @default.
• W2955275274 cites W2512791245 @default.
• W2955275274 cites W2513716142 @default.
• W2955275274 cites W2522982228 @default.
• W2955275274 cites W2548789381 @default.
• W2955275274 cites W2564062164 @default.
• W2955275274 cites W2598859771 @default.
• W2955275274 cites W2640452423 @default.
• W2955275274 cites W2732876057 @default.
• W2955275274 cites W2765167648 @default.
• W2955275274 cites W2792230432 @default.
• W2955275274 cites W2800716173 @default.
• W2955275274 cites W2802965191 @default.
• W2955275274 cites W2804543840 @default.
• W2955275274 cites W2810016976 @default.
• W2955275274 cites W2884022403 @default.
• W2955275274 cites W2888252873 @default.
• W2955275274 cites W2890883571 @default.
• W2955275274 cites W306932672 @default.
• W2955275274 doi "https://doi.org/10.1016/j.jid.2019.04.030" @default.
• W2955275274 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/31278904" @default.
• W2955275274 hasPublicationYear "2020" @default.
• W2955275274 type Work @default.
• W2955275274 sameAs 2955275274 @default.
• W2955275274 citedByCount "40" @default.
• W2955275274 countsByYear W29552752742019 @default.
• W2955275274 countsByYear W29552752742020 @default.
• W2955275274 countsByYear W29552752742021 @default.
• W2955275274 countsByYear W29552752742022 @default.
• W2955275274 countsByYear W29552752742023 @default.
• W2955275274 crossrefType "journal-article" @default.
• W2955275274 hasAuthorship W2955275274A5032128238 @default.
• W2955275274 hasAuthorship W2955275274A5036542646 @default.
• W2955275274 hasAuthorship W2955275274A5051434350 @default.
• W2955275274 hasAuthorship W2955275274A5061827333 @default.
• W2955275274 hasAuthorship W2955275274A5080087674 @default.
• W2955275274 hasAuthorship W2955275274A5080480919 @default.

• next