Anti-Inflammatory Effects Induced by Near-Infrared Light Irradiation through M2 Macrophage Polarization

Near-infrared (NIR) can penetrate the dermis. NIR is able to regulate cutaneous component cells and immune shows significant anti-inflammatory therapeutic effects. However, mechanisms of these effects are largely unknown. The purpose this study elucidate NIR-induced molecular on macrophages because play initial roles in directing responses by their M1 or M2 polarizations. Proteomic analysis revealed that radiation enhanced expression mitochondrial respiratory gene citrate synthase. This increased synthase was triggered H3K4 hypermethylation promoter but not heat, which led macrophage polarization finally resulted TGF?1 release from CD4+ cells. These cellular were validated human primary abdominal NIR-irradiated mouse experiments. In a phorbol 12-myristate 13-acetate?induced inflammatory model ear, we confirmed irradiation induced through decreased counts, reduced TNF-?, CCL22 and/or levels. Photobiomodulation (PBM) widely accepted method utilizes nonionizing visual near-infrared (700–1,100 nm) light stimulate cell functions. PBM therapy improvements wound healing, pain relief, inflammation reduction (Hamblin, 2017Hamblin M.R. Mechanisms applications photobiomodulation.AIMS Biophys. 2017; 4: 337-361Crossref PubMed Scopus (315) Google Scholar). Among PBM, anti-inflammation most reproducible event, especially improving arthritis, traumatic injuries, skin inflammation, (Avci et al., 2013Avci P. Gupta A. Sadasivam M. Vecchio D. Pam Z. N. al.Low-level laser (light) (LLLT) skin: stimulating, restoring.Semin Cutan Med Surg. 2013; 32: 41-52PubMed Scholar; Darné 2011Darné S. Hiscutt E.L. Seukeran D.C. Evaluation clinical efficacy 1,450 nm acne vulgaris: randomized split-face, investigator-blinded trial.Br J Dermatol. 2011; 165: 1256-1262Crossref (23) Hamblin, Infrared (IR) categorized into (760–1,440 nm), middle-IR (1,440–3,000 far-IR (3,000 nm–1 mm) spectra. dermis (about 5 mm depth), has biological effect cells, whereas felt as heat without penetration (Sobanko Alster, 2008Sobanko J.F. Alster T.S. Efficacy low-level for chronic ulceration humans: review discussion.Dermatol 2008; 34: 991-1000PubMed Previous studies showed an effective dosage 30–360 J/cm2 induces physiological changes epidermis, dermis, variety including fibroblasts endothelial (Dupont 2013Dupont E. Gomez J. Bilodeau Beyond UV radiation: under challenge.Int Cosmet Sci. 35: 224-232Crossref (97) Schroeder 2008Schroeder Lademann Darvin M.E. Stege H. Marks C. Bruhnke al.Infrared radiation-induced matrix metalloproteinase implications protection.J Invest 128: 2491-2497Abstract Full Text PDF (165) known induce ROS generation, leading DNA damage metalloproteinases keratinocytes well VEGF- TGF?1-associated angiogenic (Cho 2009Cho Shin M.H. Kim Y.K. Seo J.E. Lee Y.M. Park C.H. al.Effects infrared aging vivo.J Investig Dermatol Symp Proc. 2009; 14: 15-19Abstract (126) It been reported alters phenotypes markers (de Brito Sousa 2020de K. Rodrigues M.F.S.D. de Souza Santos Mesquita-Ferrari R.A. Nunes F.D. Fátima Teixeira da Silva al.Differential mediators after photobiomodulation with red lasers.Lasers 2020; 337-343Crossref (12) Fernandes 2015Fernandes K.P. N.H. Dde F. Rocha L.A. Alves A.N. al.Photobiomodulation 660-nm 780-nm activated J774 macrophage-like cells: markers.J Photochem Photobiol B. 2015; 153: 344-351Crossref (37) Our previous NIR, independent its thermal effects, enhances activation epidermal Langerhans both T helper (Th) 1 Th2 responses, indicating could host immunity (Lee 2016Lee Hong Liao W.T. Yu H.S. Differential immunological associated 2016; 155: 98-103Crossref (5) parallel, often used medical treatment, wounds, psoriasis, alopecia areata, damaged muscle tissue, pain, brain injury Most pathological events immune- inflammation-associated disorders. mechanistic less understood; therefore, focused regulation target investigate study. We customized spectrum (700–1,000 irradiator maintained constant temperature examine selected investigation responses. Macrophages plastic type hematopoietic systems. They found tissues, skin, show great functional diversity (Biswas Mantovani, 2010Biswas S.K. Mantovani Macrophage plasticity interaction lymphocyte subsets: cancer paradigm.Nat Immunol. 2010; 11: 889-896Crossref (2378) phenotypically functionally polarize switch between (classical) (alternative) subtypes differentially expressing unique surface panels mediators, cytokines, chemokines, metabolites (Shapouri-Moghaddam 2018Shapouri-Moghaddam Mohammadian Vazini Taghadosi Esmaeili S.A. Mardani al.Macrophage plasticity, polarization, function health disease.J Cell Physiol. 2018; 233: 6425-6440Crossref (1238) Therefore, functions key regulator homeostasis outcomes, dysregulation, metabolic diseases, cancers (Sica 2012Sica polarization: vivo veritas.J Clin Invest. 2012; 122: 787-795Crossref (3554) exhibit proinflammatory phenotype associate Th1 (Noël 2004Noël W. Raes G. Hassanzadeh Ghassabeh De Baetselier Beschin Alternatively during parasite infections.Trends Parasitol. 2004; 20: 126-133Abstract (234) Scholar), allergic, anti-inflammatory, immune-regulatory Th2, Th17, regulatory (Mantovani 2004Mantovani Sica Sozzani Allavena Vecchi Locati chemokine system diverse forms polarization.Trends 25: 677-686Abstract (4194) Recent evidence indicates adaptation (M1 polarization) (Diskin Pålsson-McDermott, 2018Diskin Pålsson-McDermott E.M. Metabolic modulation effector function.Front 9: 270Crossref (139) pathways include glycolysis, citric acid cycle, oxidative phosphorylation (adenosine triphosphate [ATP] generation), production (O’Neill Hardie, 2013O’Neill Hardie D.G. Metabolism limited AMPK pseudo-starvation.Nature. 493: 346-355Crossref (748) Pelegrin Surprenant, 2009Pelegrin Surprenant Dynamics reveal new mechanism inhibit IL-1beta pyrophosphates.EMBO 28: 2114-2127Crossref (204) Because it mitochondria photoacceptors visible IR (Karu, 2008Karu T.I. Mitochondrial signaling mammalian near-IR radiation.Photochem Photobiol. 84: 1091-1099Crossref (358) Karu 2004Karu Pyatibrat L.V. Kalendo G.S. Photobiological attachment via cytochrome c oxidase.Photochem 3: 211-216Crossref (132) hypothesized might alter mitochondria-associated pathways, aberrant polarization. To test our hypothesis, standardized THP-1 monocyte line discovery phase mice models validate macrophages. generates 700–1,000 (Supplementary Figure S1a). Cytotoxicity tests indicated 30 60 treatments had no cytotoxic 24 hours. contrast, 90 120 S1b) apoptosis hours S1c). treatment (10-minute irradiation) further identify possible molecules affected performed shotgun quantitative proteomic analysis. There 226 proteins identified NIR-treated nine significantly eight (Table 1). these, CS protein proteomics profile S2). Using Ingenuity Pathway Analysis bioinformatics software (Qiagen, Chatsworth, CA), three major altered categories, death survival, energy production, small molecule biochemistry, NIR-exposed involved categories CS, EF1A1, transitional endoplasmic reticulum ATPase (also VCP), ATP5A1 S3). mRNA genes using real-time PCR. (60 J/cm2) expressions medium 37 °C 39 °C, tested (without NIR) expression. results did (Figure 1a). increase western blotting 1b). involvement epigenetic expression, histone acetyltransferase inhibitor anacardic acid, methyltransferase 5?-methylthioadenosine (MTA), deacetylase trichostatin A used. data be inhibited MTA 1c). Furthermore, activity S4). checked methylation sites gene. trimethylation (but H3K9 trimethylation) 1d). heat-treated 1e). heat) promoter.Table 1Significantly Increased Decreased Proteins Cells Treatment AnalysisGene SymbolFold ChangeMassSDHitScoreDescriptionCS51.3152,6365.009,364Citrate synthase, mitochondrialKIAA154912.36212,88711.0110,556UPF0606 KIAA1549PKDCC3.4655,2735.4921,321Protein kinase domain?containing protein, cytoplasmicEF1A13.3051,7696.977,484Elongation factor 1-?1VCP2.7291,45712.8842,164Transitional ATPaseK92.1063,0881.1121,276Keratin, I cytoskeletal 9K12.0066,9551.097,285Keratin, II 1ATP5A11.8960,7892.659,861ATP subunit ?, mitochondrialEPRS1.68176,2283.7613,440Bifunctional aminoacyl-tRNA synthetaseGLO1?2.0021,4971.3925,262Lactoylglutathione lyasePSMA4?2.0230,4791.896,794Proteasome ? type-4HIST1H4A?2.0311,6691.0629,221Histone H4IDI1?2.0727,1771.7619,028Isopentenyl-diphosphate ?-isomerase 1ALOX5?2.1918,3721.2718,928Arachidonate 5-lipoxygenase-activating proteinUGGG1?2.48180,6782.2410,258UDP-glucose: glycoprotein glucosyltransferase 1SOX30?5.2282,98818.8510,158Transcription SOX-30TLN1?8.53276,95914.7523,266Talin-1Abbreviations: ATP, adenosine triphosphate; K, keratin; tRNA, transfer RNA; UDP, uridine diphosphate. Open table tab Abbreviations: CXCL10 marker, M2a CCL1 M2b CCL16 M2c marker (Benoit 2008Benoit Desnues Mege J.L. bacterial infections.J 181: 3733-3739Crossref (876) ELISA level 2a). Similar observed PCR, shown Supplementary S5. flow cytometric CD80 (for M1) CD206 M2) staining 2b). 13-acetate (PMA)-primed Results overexpression PMA-primed either interfering RNA 2c) 2d). reversed S6). irradiation. collected cell-free?conditioned soluble factors THP-1?conditioned (Th cells). condition medium, cytokines IFN-?, IL-4, TGF?1, IL-17A compared those non-NIR?irradiated control S7). When treated cells?associated cytokine Th1-associated Th2-associated Th17-associated 2e). Conditioned NIR?treated 2f). Similarly, conditioned RNA?pretreated 2g). neutralization antibodies release. antibody S8). Monocyte-derived (MDMs) spindle dendritic morphology CD80+ markers. After irradiation, MDM CD206+ reprogram S9). result S10). MDM, 3a) expression; 3b). (27.2 ± 4.8 pg/ml) untreated (12.0 2.4 pg/ml). 3c). cocultured MDMs isolated (from same donor) transwell 0.22 ?M pore size. contact system, although pass pores. MDM?CD4+ coculture, pretreatment 3d). alteration event also irradiated cultured intraperitoneal C57BL/6 (B6) NIR. studied modification regulations peritoneal 3e) 3f). Functionally, 3g). transwell, 3h). confirm B6 2 hours, count lavage and, at time, cytometry S11a). CXCL10, CCL22, levels lipopolysaccharide-pretreated (stimulatory effect) Figures S11b?d). penetrates M2-promoting ear tissue PMA (Silván 1996Silván A.M. Abad M.J. Bermejo Villar Inhibition hydroxyachillin, sesquiterpene lactone Tanacetum microphyllum, PMA-induced oedema.Inflamm Res. 1996; 45: 289-292Crossref (15) redness 4a) swelling 4b). PMA-treated number decreased, 4c). addition, 4d). findings suggest switched ears. PMA- NIR-cotreated 4e). M2/T ELISA. TNF-? ears 4f). suggested level, skin. vivo, interference intravenous injection mice. abolished is, S12). vivo. aims terms macrophage-mediated established device animal models. study, per se causes drives toward phenotype. gel-free analysis, upregulated essential enzymatic tricarboxylic cycle. Tricarboxylic cycle?associated reprogramming one dominant (Haschemi 2012Haschemi Kosma Gille L. Evans C.R. Burant C.F. Starkl al.The sedoheptulose CARKL directs glucose metabolism.Cell Metab. 15: 813-826Abstract (354) Kelly O’Neill, 2015Kelly O’Neill innate immunity.Cell 771-784Crossref (742) More specifically, necessary IL-4?induced 2011Cho S.H. Ahn A.K. Bhargava Eischen C.M. McGuinness O. al.Glycolytic rate lymphomagenesis depend PARP14, ADP ribosyltransferase B aggressive lymphoma (BAL) family.Proc Natl Acad Sci USA. 108: 15972-15977Crossref (78) regulated °C. activate transcription (Gupta 2010Gupta S.Y. Artis Molfese D.L. Schumacher Sweatt J.D. al.Histone regulates memory formation.J Neurosci. 30: 3589-3599Crossref (424) line, macrophage, macrophages, Shiurba 2006Shiurba R. Hirabayashi T. Masuda Kawamura Komoike Y. Klitz al.Cellular ciliate, Tetrahymena thermophila, far irradiation.Photochem 2006; 5: 799-807Crossref (10) Scholar thermophile, growth. imply IR-induced common phenomena single-celled organisms epidermal-resident heating alone. dendrite formation numbers skin-draining lymph nodes sheets. investigated dermal switching decrease THP-1, culture tests. first analyze M1/M2 switching. Abdominal count, lavage. demonstrates inducer cause M1-dominate (with TNF-?) events, resulting inhibition abolish understood. only clearly epigenetically leads TGF?1.