Top

Published in:

20-01-2023 | Diabetic Foot | Original Article

Pterostilbene attenuates hemin-induced dysregulation of macrophage M2 polarization via Nrf2 activation in experimental hyperglycemia

Authors: Goutham V. Ganesh, Kunka Mohanram Ramkumar

Published in: Inflammopharmacology | Issue 4/2023

Abstract

Macrophages exhibit a high degree of plasticity that is physiologically relevant in wound healing, and disruption in normal macrophage response leads to delayed wound closure resulting in chronic wounds. Here, we attempt to discern macrophage responses to hemin via regulation of the nuclear factor-erythroid factor 2-related factor 2 (Nrf2) that could help us better understand the pathophysiology of diabetic foot ulcers (DFU). We demonstrate the alleviation of hemin-mediated Nrf2 suppression and M2 macrophage polarization by pterostilbene (PTS), a proven Nrf2 activator. IC-21 macrophages were treated with hemin under the normoglycemic or hyperglycemic environment with or without PTS and the expression levels of various markers, such as Nrf2 and its downstream target Heme Oxygenase-1 (HO-1), CD206, Ferroportin-1 among others were analyzed using qPCR and Western blot. Our results revealed that hemin under hyperglycemia reduced Nrf2 activation and its downstream targets, M2 polarization, and the induction of a proinflammatory cellular environment, and interestingly all of these were remedied by PTS treatment. Gelatin zymography of matrix metalloproteinase2 (MMP2) expression revealed that hemin under hyperglycemic condition significantly elevated MMP2 expression, which was reversed by PTS treatment. Further proteomic analysis using liquid chromatography with tandem mass spectrometry (LC–MS/MS) revealed a heightened cellular stress profile accompanying inflammation that was suppressed by PTS. This study has furthered our understanding on the role of Nrf2 in attenuating hemin-induced perturbations in macrophage responses and suggests a potential therapeutic target in the management of DFU.

Abdalla HB, Napimoga MH, Lopes AH, de Macedo Maganin AG, Cunha TM, Van Dyke TE, Clemente Napimoga JT (2020) Activation of PPAR-gamma induces macrophage polarization and reduces neutrophil migration mediated by heme oxygenase 1. Int Immunopharmacol 84:106565. https://doi.org/10.1016/j.intimp.2020.106565CrossRefPubMed

Allhorn M, Lundqvist K, Schmidtchen A, Akerstrom B (2003) Heme-scavenging role of alpha1-microglobulin in chronic ulcers. J Invest Dermatol 121(3):640–646. https://doi.org/10.1046/j.1523-1747.2003.12409.xCrossRefPubMed

Cada DJ, Levien TL, Baker DE (2013) Dimethyl fumarate. Hosp Pharm 48(8):668–679. https://doi.org/10.1310/hpj4808-668CrossRefPubMedPubMedCentral

Chamberlain LM, Godek ML, Gonzalez-Juarrero M, Grainger DW (2009) Phenotypic non-equivalence of murine (monocyte-) macrophage cells in biomaterial and inflammatory models. J Biomed Mater Res A 88(4):858–871. https://doi.org/10.1002/jbm.a.31930CrossRefPubMedPubMedCentral

Chen QY, Wang GG, Li W, Jiang YX, Lu XH, Zhou PP (2016) Heme oxygenase-1 promotes delayed wound healing in diabetic rats. J Diabetes Res 2016:9726503. https://doi.org/10.1155/2016/9726503CrossRefPubMed

Choi KM, Kashyap PC, Dutta N, Stoltz GJ, Ordog T, Shea Donohue T et al (2010) CD206-positive M2 macrophages that express heme oxygenase-1 protect against diabetic gastroparesis in mice. Gastroenterology 138(7):2399-2409 e2391. https://doi.org/10.1053/j.gastro.2010.02.014CrossRefPubMed

Corna G, Campana L, Pignatti E, Castiglioni A, Tagliafico E, Bosurgi L et al (2010) Polarization dictates iron handling by inflammatory and alternatively activated macrophages. Haematologica 95(11):1814–1822. https://doi.org/10.3324/haematol.2010.023879CrossRefPubMedPubMedCentral

Dinh T, Tecilazich F, Kafanas A, Doupis J, Gnardellis C, Leal E et al (2012) Mechanisms involved in the development and healing of diabetic foot ulceration. Diabetes 61(11):2937–2947. https://doi.org/10.2337/db12-0227CrossRefPubMedPubMedCentral

Dornadula S, Thiruppathi S, Palanisamy R, Umapathy D, Suzuki T, Ramkumar KM (2019) Differential proteomic profiling identifies novel molecular targets of pterostilbene against experimental diabetes. J Cell Physiol 234(3):1996–2012. https://doi.org/10.1002/jcp.26835CrossRefPubMed

Ganesh GV, Ramkumar KM (2021) Dysregulation of Nrf2 redox pathway in macrophages under diabetic microenvironment. Exp Gerontol 152:111479. https://doi.org/10.1016/j.exger.2021.111479CrossRefPubMed

Hozzein WN, Badr G, Badr BM, Allam A, Ghamdi AA, Al-Wadaan MA, Al-Waili NS (2018) Bee venom improves diabetic wound healing by protecting functional macrophages from apoptosis and enhancing Nrf2, Ang-1 and Tie-2 signaling. Mol Immunol 103:322–335. https://doi.org/10.1016/j.molimm.2018.10.016CrossRefPubMed

Imoto S, Kono M, Suzuki T, Shibuya Y, Sawamura T, Mizokoshi Y et al (2018) Haemin-induced cell death in human monocytic cells is consistent with ferroptosis. Transfus Apher Sci 57(4):524–531. https://doi.org/10.1016/j.transci.2018.05.028CrossRefPubMed

Joshi N, Werner S (2017) Nrf2 is highly expressed in neutrophils, but myeloid cell-derived Nrf2 is dispensable for wound healing in mice. PLoS ONE 12(10):e0187162. https://doi.org/10.1371/journal.pone.0187162CrossRefPubMedPubMedCentral

Kallenborn-Gerhardt W, Moser CV, Lorenz JE, Steger M, Heidler J, Scheving R et al (2017) Rab7-a novel redox target that modulates inflammatory pain processing. Pain 158(7):1354–1365. https://doi.org/10.1097/j.pain.0000000000000920CrossRefPubMed

Kumar D, Jena GR, Ram M, Lingaraju MC, Singh V, Prasad R et al (2019) Hemin attenuated oxidative stress and inflammation to improve wound healing in diabetic rats. Naunyn Schmiedebergs Arch Pharmacol 392(11):1435–1445. https://doi.org/10.1007/s00210-019-01682-7CrossRefPubMed

Lee YJ, Kwon SB, An JM, Kim CH, Lee SH, Choi CY et al (2015) Increased protein oxidation and decreased expression of nuclear factor E2-related factor 2 protein in skin tissue of patients with diabetes. Clin Exp Dermatol 40(2):192–200. https://doi.org/10.1111/ced.12487CrossRefPubMed

Li Y, Ma F, Li H, Song Y, Zhang H, Jiang Z, Wu H (2018) Dimethyl fumarate accelerates wound healing under diabetic condition. J Mol Endocrinol 61(4):163–172. https://doi.org/10.1530/JME-18-0102CrossRefPubMed

Li M, Yu H, Pan H, Zhou X, Ruan Q, Kong D et al (2019) Nrf2 suppression delays diabetic wound healing through sustained oxidative stress and inflammation. Front Pharmacol 10:1099. https://doi.org/10.3389/fphar.2019.01099CrossRefPubMedPubMedCentral

Lu J, Xie L, Liu C, Zhang Q, Sun S (2017) PTEN/PI3k/AKT regulates macrophage polarization in emphysematous mice. Scand J Immunol 85(6):395–405. https://doi.org/10.1111/sji.12545CrossRefPubMed

Marro S, Chiabrando D, Messana E, Stolte J, Turco E, Tolosano E, Muckenthaler MU (2010) Heme controls ferroportin1 (FPN1) transcription involving Bach1, Nrf2 and a MARE/ARE sequence motif at position -7007 of the FPN1 promoter. Haematologica 95(8):1261–1268. https://doi.org/10.3324/haematol.2009.020123CrossRefPubMedPubMedCentral

Nakaso K, Yano H, Fukuhara Y, Takeshima T, Wada-Isoe K, Nakashima K (2003) PI3K is a key molecule in the Nrf2-mediated regulation of antioxidative proteins by hemin in human neuroblastoma cells. FEBS Lett 546(2–3):181–184. https://doi.org/10.1016/s0014-5793(03)00517-9CrossRefPubMed

Qiu-Yi Choo SCMY, Ho PC, Tanaka Y, Lin H-S (2014) Pterostilbene surpassed resveratrol for anti-inflammatory application: potency consideration and pharmacokinetics perspective. J Funct Foods 11:352–362. https://doi.org/10.1016/j.jff.2014.10.018CrossRef

Ram M, Singh V, Kumawat S, Kumar D, Lingaraju MC, Uttam Singh T et al (2015) Deferoxamine modulates cytokines and growth factors to accelerate cutaneous wound healing in diabetic rats. Eur J Pharmacol 764:9–21. https://doi.org/10.1016/j.ejphar.2015.06.029CrossRefPubMed

Ramkumar KM, Sekar TV, Foygel K, Elango B, Paulmurugan R (2013) Reporter protein complementation imaging assay to screen and study Nrf2 activators in cells and living animals. Anal Chem 85(15):7542–7549. https://doi.org/10.1021/ac401569jCrossRefPubMedPubMedCentral

Safety and Efficacy of ActiGraft Compared to Standard of Care in DFUs (2019) Clinical Trials. Retrieved 30 November 2021, from https://clinicaltrials.gov/ct2/show/NCT04185558

Schaer CA, Schoedon G, Imhof A, Kurrer MO, Schaer DJ (2006) Constitutive endocytosis of CD163 mediates hemoglobin-heme uptake and determines the noninflammatory and protective transcriptional response of macrophages to hemoglobin. Circ Res 99(9):943–950. https://doi.org/10.1161/01.RES.0000247067.34173.1bCrossRefPubMed

Sindrilaru A, Peters T, Wieschalka S, Baican C, Baican A, Peter H et al (2011) An unrestrained proinflammatory M1 macrophage population induced by iron impairs wound healing in humans and mice. J Clin Invest 121(3):985–997. https://doi.org/10.1172/JCI44490CrossRefPubMedPubMedCentral

Sireesh D, Ganesh MR, Dhamodharan U, Sakthivadivel M, Sivasubramanian S, Gunasekaran P, Ramkumar KM (2017) Role of pterostilbene in attenuating immune mediated devastation of pancreatic beta cells via Nrf2 signaling cascade. J Nutr Biochem 44:11–21. https://doi.org/10.1016/j.jnutbio.2017.02.015CrossRefPubMed

Soares MA, Cohen OD, Low YC, Sartor RA, Ellison T, Anil U et al (2016) Restoration of Nrf2 signaling normalizes the regenerative niche. Diabetes 65(3):633–646. https://doi.org/10.2337/db15-0453CrossRefPubMed

Song L, Lee C, Schindler C (2011) Deletion of the murine scavenger receptor CD68. J Lipid Res 52(8):1542–1550. https://doi.org/10.1194/jlr.M015412CrossRefPubMedPubMedCentral

Suganya N, Bhakkiyalakshmi E, Subin TS, Krishnamurthi K, Devi SS, Lau K et al (2014) Proteomic identification of pterostilbene-mediated anticancer activities in HepG2 cells. Chem Res Toxicol 27(7):1243–1252. https://doi.org/10.1021/tx5001392CrossRefPubMed

Takahashi Y, Kobayashi T, D’Alessandro-Gabazza CN, Toda M, Fujiwara K, Okano T et al (2019) Protective role of matrix metalloproteinase-2 in allergic bronchial asthma. Front Immunol 10:1795. https://doi.org/10.3389/fimmu.2019.01795CrossRefPubMedPubMedCentral

Tchanque-Fossuo C, Dahle SE, Buchman S, Isseroff R (2016) 737 Deferoxamine in wound healing. J Investig Dermatol 136:S130. https://doi.org/10.1016/j.jid.2016.02.780CrossRef

Toby Richards SH, Wright J, Loukogeorgakis S, Khanbhai M (2012) Anaemia, inflammation, renal function, and the diabetic foot: What are the relationships? Diabetic Foot J 15(4):150–158

Vella L, Gatt A, Formosa C (2017) Does baseline hemoglobin a1c level predict diabetic foot ulcer outcome or wound healing time? J Am Podiatr Med Assoc 107(4):272–279. https://doi.org/10.7547/15-176CrossRefPubMed

Vijayan V, Wagener F, Immenschuh S (2018) The macrophage heme-heme oxygenase-1 system and its role in inflammation. Biochem Pharmacol 153:159–167. https://doi.org/10.1016/j.bcp.2018.02.010CrossRefPubMed

Wang WT, Sun L, Sun CH (2019) PDIA3-regulated inflammation and oxidative stress contribute to the traumatic brain injury (TBI) in mice. Biochem Biophys Res Commun 518(4):657–663. https://doi.org/10.1016/j.bbrc.2019.08.100CrossRefPubMed

Zhang F, Fan D, Mo XN (2018) Prohibitin and the extracellular matrix are upregulated in murine alveolar epithelial cells with LPS-induced acute injury. Mol Med Rep 17(6):7769–7773. https://doi.org/10.3892/mmr.2018.8808CrossRefPubMed

Zhao S, Ghosh A, Lo CS, Chenier I, Scholey JW, Filep JG et al (2018) Nrf2 deficiency upregulates intrarenal angiotensin-converting enzyme-2 and angiotensin 1–7 receptor expression and attenuates hypertension and nephropathy in diabetic mice. Endocrinology 159(2):836–852. https://doi.org/10.1210/en.2017-00752CrossRefPubMed

Zhou J, Ci X, Ma X, Yu Q, Cui Y, Zhen Y, Li S (2019) Pterostilbene activates the Nrf2-dependent antioxidant response to ameliorate arsenic-induced intracellular damage and apoptosis in human keratinocytes. Front Pharmacol 10:497. https://doi.org/10.3389/fphar.2019.00497CrossRefPubMedPubMedCentral

Title: Pterostilbene attenuates hemin-induced dysregulation of macrophage M2 polarization via Nrf2 activation in experimental hyperglycemia
Authors: Goutham V. Ganesh
Kunka Mohanram Ramkumar
Publication date: 20-01-2023
Publisher: Springer International Publishing
Keywords: Diabetic Foot
Hyperglycemia
Published in: Inflammopharmacology / Issue 4/2023
Print ISSN: 0925-4692
Electronic ISSN: 1568-5608
DOI: https://doi.org/10.1007/s10787-023-01134-y

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Pterostilbene attenuates hemin-induced dysregulation of macrophage M2 polarization via Nrf2 activation in experimental hyperglycemia

Abstract

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Please log in to get access to this content

Other articles of this Issue 4/2023

Anakinra for the prophylaxis of venous thromboembolism in patients with COVID-19

Effect of ethyl gallate and propyl gallate on dextran sulfate sodium (DSS)-induced ulcerative colitis in C57BL/6 J mice: preventive and protective

Fractionated whole body γ-irradiation aggravates arthritic severity via boosting NLRP3 and RANKL expression in adjuvant-induced arthritis model: the mitigative potential of ebselen

Protective effects of Stephania pierrei tuber-derived oxocrebanine against LPS-induced acute lung injury in mice

The potential therapeutic effect of statins in multiple sclerosis: beneficial or detrimental effects

Ischemic tubular injury: Oxygen-sensitive signals and metabolic reprogramming