{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T04:33:49Z","timestamp":1787027629098,"version":"build-2736575974"},"reference-count":52,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2023,5,13]],"date-time":"2023-05-13T00:00:00Z","timestamp":1683936000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000053","name":"National Eye Institute","doi-asserted-by":"publisher","award":["R21EY031031"],"award-info":[{"award-number":["R21EY031031"]}],"id":[{"id":"10.13039\/100000053","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biomolecules"],"abstract":"<jats:p>Purpose: To investigate the anti-inflammatory and anti-angiogenic effects of the bioactive lipid mediator LXA4 on a rat model of severe corneal alkali injury. Methods: To induce a corneal alkali injury in the right eyes of anesthetized Sprague Dawley rats. They were injured with a \u03a6 4 mm filter paper disc soaked in 1 N NaOH placed on the center of the cornea. After injury, the rats were treated topically with LXA4 (65 ng\/20 \u03bcL) or vehicle three times a day for 14 days. Corneal opacity, neovascularization (NV), and hyphema were recorded and evaluated in a blind manner. Pro-inflammatory cytokine expression and genes involved in cornel repair were assayed by RNA sequencing and capillary Western blot. Cornea cell infiltration and monocytes isolated from the blood were analyzed by immunofluorescence and by flow cytometry. Results: Topical treatment with LXA4 for two weeks significantly reduced corneal opacity, NV, and hyphema compared to the vehicle treatment. RNA-seq and Western blot results showed that LXA4 decreased the gene and protein expression of pro-inflammatory cytokines interleukin (IL)-1\u03b2 and IL-6 and pro-angiogenic mediators matrix metalloproteinase (MMP)-9 and vascular endothelial growth factor (VEGFA). It also induces genes involved in keratinization and ErbB signaling and downregulates immune pathways to stimulate wound healing. Flow cytometry and immunohistochemistry showed significantly less infiltration of neutrophils in the corneas treated with LXA4 compared to vehicle treatment. It also revealed that LXA4 treatment increases the proportion of type 2 macrophages (M2) compared to M1 in blood-isolated monocytes. Conclusions: LXA4 decreases corneal inflammation and NV induced by a strong alkali burn. Its mechanism of action includes inhibition of inflammatory leukocyte infiltration, reduction in cytokine release, suppression of angiogenic factors, and promotion of corneal repair gene expression and macrophage polarization in blood from alkali burn corneas. LXA4 has potential as a therapeutic candidate for severe corneal chemical injuries.<\/jats:p>","DOI":"10.3390\/biom13050831","type":"journal-article","created":{"date-parts":[[2023,5,15]],"date-time":"2023-05-15T02:56:57Z","timestamp":1684119417000},"page":"831","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":18,"title":["Lipoxin A4 (LXA4) Reduces Alkali-Induced Corneal Inflammation and Neovascularization and Upregulates a Repair Transcriptome"],"prefix":"10.3390","volume":"13","author":[{"given":"Jiucheng","family":"He","sequence":"first","affiliation":[{"name":"Neuroscience Center of Excellence, School of Medicine, Louisiana State University Health New Orleans, New Orleans, LA 70112, USA"},{"name":"Department of Ophthalmology, School of Medicine, Louisiana State University Health New Orleans, New Orleans, LA 70112, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thang L.","family":"Pham","sequence":"additional","affiliation":[{"name":"Neuroscience Center of Excellence, School of Medicine, Louisiana State University Health New Orleans, New Orleans, LA 70112, USA"},{"name":"HENIKAA Research and Technology Institute (PRATI), A&A Green Phoenix Group JSC, Hanoi 11313, Vietnam"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Azucena H.","family":"Kakazu","sequence":"additional","affiliation":[{"name":"Neuroscience Center of Excellence, School of Medicine, Louisiana State University Health New Orleans, New Orleans, LA 70112, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Abhilash","family":"Ponnath","sequence":"additional","affiliation":[{"name":"Neuroscience Center of Excellence, School of Medicine, Louisiana State University Health New Orleans, New Orleans, LA 70112, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Khanh V.","family":"Do","sequence":"additional","affiliation":[{"name":"Faculty of Medicine, PHENIKAA University, Hanoi 12116, Vietnam"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7172-7870","authenticated-orcid":false,"given":"Haydee E. P.","family":"Bazan","sequence":"additional","affiliation":[{"name":"Neuroscience Center of Excellence, School of Medicine, Louisiana State University Health New Orleans, New Orleans, LA 70112, USA"},{"name":"Department of Ophthalmology, School of Medicine, Louisiana State University Health New Orleans, New Orleans, LA 70112, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,5,13]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"275","DOI":"10.1016\/S0039-6257(96)00007-0","article-title":"Chemical injuries of the eye: Current concepts in pathophysiology and therapy","volume":"41","author":"Wagoner","year":"1997","journal-title":"Surv. Ophthalmol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"2196","DOI":"10.1016\/S0161-6420(00)00390-0","article-title":"The prevalence and 5-year incidence of ocular trauma: The beaver dam eye study","volume":"107","author":"Wong","year":"2000","journal-title":"Ophthalmology"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"48","DOI":"10.1016\/j.jtos.2016.09.002","article-title":"Current and Upcoming Therapies for Ocular Surface Chemical Injuries","volume":"15","author":"Eslani","year":"2017","journal-title":"Ocul. Surf."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"100033","DOI":"10.1194\/jlr.TR120000954","article-title":"Docosanoids signaling modulates corneal nerve regeneration: Effect on tear secretion, wound healing and neuropathic pain","volume":"62","author":"Pham","year":"2021","journal-title":"J. Lipid Res."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"705","DOI":"10.1136\/bjo.66.11.705","article-title":"Corticosteroids and corneal epithelial wound healing","volume":"66","author":"Petroutsos","year":"1982","journal-title":"Br. J. Ophthalmol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"884","DOI":"10.1001\/jama.1963.03060110088027","article-title":"Elevated intraocular pressure following corticosteroid eye drops","volume":"185","author":"Becker","year":"1963","journal-title":"JAMA"},{"key":"ref_7","first-page":"570","article-title":"Prevention of stromal ulceration in the alkali-burned rabbit cornea by glued-on contact lens. Evidence for the role of polymorphonuclear leukocytes in collagen degradation","volume":"18","author":"Kenyon","year":"1979","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"431","DOI":"10.1001\/archopht.116.4.431","article-title":"Amniotic membrane transplantation with or without limbal allografts for corneal surface reconstruction in patients with limbal stem cell deficiency","volume":"116","author":"Tseng","year":"1998","journal-title":"Arch. Ophthalmol."},{"key":"ref_9","first-page":"1526","article-title":"The efficacy of ascorbate treatment after severe experimental alkali burns de-pends upon the route of administration","volume":"19","author":"Pfister","year":"1980","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_10","first-page":"107","article-title":"Inhibition of alkali-induced corneal ulceration and perforation by a thiol peptide","volume":"31","author":"Burns","year":"1990","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_11","first-page":"1340","article-title":"Effect of synthetic metalloproteinase inhibitor or citrate on neutrophil chemotaxis and the respiratory burst","volume":"38","author":"Pfister","year":"1997","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_12","first-page":"2430","article-title":"Effect of metalloproteinase inhibitor on corneal cytokine expression after alkali injury","volume":"40","author":"Sotozono","year":"1999","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1097\/ICO.0000000000000534","article-title":"Management of limbal stem cells deficiency: A historical perspective, past, present and future","volume":"34","author":"Holland","year":"2015","journal-title":"Cornea"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"5124","DOI":"10.1111\/jcmm.16501","article-title":"Priming human adipose-derived mesenchymal stem cells for corneal surface regeneration","volume":"25","year":"2021","journal-title":"J. Cell. Mol. Med."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"18","DOI":"10.1016\/j.trsl.2018.11.003","article-title":"A proof-of-concept clinical trial using mesenchymal stem cells for the treatment of corneal epithelial stem cell deficiency","volume":"206","author":"Calonge","year":"2018","journal-title":"Transl. Res."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"726","DOI":"10.1038\/nri2395","article-title":"Mesenchymal stem cells in health and disease","volume":"8","author":"Uccelli","year":"2008","journal-title":"Nat. Rev. Immunol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"134","DOI":"10.1080\/08820538.2017.1353834","article-title":"Limbal stem cell transplantation and complications","volume":"33","author":"Yin","year":"2017","journal-title":"Semin. Ophthalmol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"481","DOI":"10.1016\/0014-4835(91)90046-H","article-title":"Platelet-activating factor (PAF) accumulation correlates with injury in the cornea","volume":"52","author":"Bazan","year":"1991","journal-title":"Exp. Eye Res."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"449","DOI":"10.1016\/S1350-9462(02)00011-3","article-title":"The role of platelet-activating factor in the corneal response to injury","volume":"21","author":"Bazan","year":"2002","journal-title":"Prog. Retin. Eye Res."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1063","DOI":"10.3109\/02713683.2010.513797","article-title":"The induction of an angiogenic response in corneal myofibroblasts by platelet-activating factor (PAF)","volume":"35","author":"He","year":"2010","journal-title":"Curr. Eye Res."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"670","DOI":"10.1076\/ceyr.16.7.670.5057","article-title":"Cytokine expression in the alkali-burned cornea","volume":"16","author":"Sotozono","year":"1997","journal-title":"Curr. Eye Res."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"101","DOI":"10.1146\/annurev.immunol.25.022106.141647","article-title":"Resolution Phase of Inflammation: Novel Endogenous Anti-Inflammatory and Proresolving Lipid Mediators and Pathways","volume":"25","author":"Serhan","year":"2007","journal-title":"Annu. Rev. Immunol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"104","DOI":"10.1016\/j.jtos.2020.04.008","article-title":"The potential of lipid mediator networks as ocular surface therapeutics and biomarkers","volume":"19","author":"Flitter","year":"2020","journal-title":"Ocul. Surf."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"235","DOI":"10.1089\/jop.2010.0187","article-title":"Aspirin-Triggered Lipoxin A4 (15-epi-LXA4) Increases the Endothelial Viability of Human Corneas Storage in Optisol-GS","volume":"27","author":"He","year":"2011","journal-title":"J. Ocul. Pharmacol. Ther."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1016\/j.exer.2012.07.008","article-title":"Lipoxin A\u2084 inhibits platelet-activating factor inflammatory response and stim-ulates corneal wound healing of injuries that compromise the stroma","volume":"103","author":"Kakazu","year":"2012","journal-title":"Exp. Eye Res."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"2240","DOI":"10.1167\/iovs.10-6199","article-title":"EGF Stimulates Lipoxin A4 Synthesis and Modulates Repair in Corneal Epithelial Cells through ERK and p38 Activation","volume":"52","author":"Kenchegowda","year":"2011","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"161","DOI":"10.1016\/S0014-4835(02)00293-2","article-title":"Local suppression of IL-1 by receptor antagonist in the rat model of corneal alkali injury","volume":"76","author":"Yamada","year":"2003","journal-title":"Exp. Eye Res."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"194","DOI":"10.1136\/bjo.2009.173724","article-title":"Comparison of prognostic value of Roper Hall and Dua classification systems in acute ocular burns","volume":"95","author":"Gupta","year":"2010","journal-title":"Br. J. Ophthalmol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1348","DOI":"10.1080\/02713683.2017.1317817","article-title":"Comprehensive Modeling of Corneal Alkali Injury in the Rat Eye","volume":"42","author":"Choi","year":"2017","journal-title":"Curr. Eye Res."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"e108","DOI":"10.1093\/nar\/gkt214","article-title":"The Subread aligner: Fast, accurate and scalable read mapping by seed-and-vote","volume":"41","author":"Liao","year":"2013","journal-title":"Nucleic Acids Res."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"923","DOI":"10.1093\/bioinformatics\/btt656","article-title":"featureCounts: An efficient general-purpose program for assigning sequence reads to genomic features","volume":"30","author":"Liao","year":"2014","journal-title":"Bioinformatics"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"550","DOI":"10.1186\/s13059-014-0550-8","article-title":"Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2","volume":"15","author":"Love","year":"2014","journal-title":"Genome Biol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"12787","DOI":"10.1038\/s41598-021-92293-x","article-title":"ELV-N32 and RcD6 isomer decrease proinflammatory cytokines, senescence programming, ACE2 and SARS-CoV-2 spike protein RBD binding in injured cornea","volume":"11","author":"Pham","year":"2021","journal-title":"Sci. Rep."},{"key":"ref_34","first-page":"e54599","article-title":"Phenotypic Characterization of Macrophages from Rat Kidney by Flow Cytometry","volume":"116","author":"Cortegano","year":"2016","journal-title":"J. Vis. Exp."},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Able, J.R., Boyles, D.A., Walters, A.W., Kujawa, M.R., McMillen, C.M., Reed, D.S., and Hartman, A.L. (2019). Neutrophil and macrophage influx into the central nervous system are inflammatory components of lethal Rift Valley fever encephalitis in rats. PLoS Pathog., 15.","DOI":"10.1371\/journal.ppat.1007833"},{"key":"ref_36","first-page":"345","article-title":"Platelet-activating factor induces the expression of metalloproteinases-1 and -9, but not -2 or -3, in the corneal epithelium","volume":"36","author":"Tao","year":"1995","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_37","first-page":"1422","article-title":"Delay of corneal epithelial wound healing and induction of keratocyte apoptosis by platelet-activating factor","volume":"43","author":"Chandrasekher","year":"2002","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"70","DOI":"10.1001\/archopht.124.1.70","article-title":"Alkali-induced corneal stromal melting prevention by a novel platelet-activating factor receptor antagonist","volume":"124","author":"He","year":"2006","journal-title":"Arch. Ophthalmol."},{"key":"ref_39","first-page":"645","article-title":"Platelet activating factor in anaphylaxis and phagocytosis. I. Release from human peripheral polymorphonuclear and monocytes during the stimulation by ionophore A23187 and phagocytosis but not from degranulating basophils","volume":"40","author":"Alonso","year":"1980","journal-title":"Immunology"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"437","DOI":"10.1111\/j.1365-2362.1980.tb02082.x","article-title":"Release of platelet-activating factor (PAF-acether) from alveolar macrophages by the calcium ionophore A 23187 and phagocytosis","volume":"10","author":"Arnoux","year":"1980","journal-title":"Eur. J. Clin. Investig."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"514","DOI":"10.3389\/fimmu.2014.00514","article-title":"From monocytes to M1\/M2 macrophages: Phenotypical vs. functional differentiation","volume":"5","author":"Italiani","year":"2014","journal-title":"Front. Immunol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"235","DOI":"10.3109\/08820538.2011.588652","article-title":"Inflammatory corneal neovascularization: Etiopathogenesis","volume":"26","author":"Clements","year":"2011","journal-title":"Semin. Ophthalmol."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"98","DOI":"10.1016\/j.exer.2012.02.015","article-title":"Anti-inflammatory effect of IL-6 receptor blockade in corneal alkali burn","volume":"97","author":"Sakimoto","year":"2012","journal-title":"Exp. Eye Res."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"227","DOI":"10.1016\/S1542-0124(11)70035-0","article-title":"Anti-VEGF Treatment of Corneal Neovascularization","volume":"9","author":"Keating","year":"2011","journal-title":"Ocul. Surf."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"485","DOI":"10.1111\/cbdd.13529","article-title":"Regulation of matrix metalloproteinases 2 and 9 in corneal neovascularization","volume":"95","author":"Zhang","year":"2019","journal-title":"Chem. Biol. Drug Des."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"4360","DOI":"10.1167\/iovs.06-1234","article-title":"Positive feedback regulation between MMP-9 and VEGF in human RPE cells","volume":"48","author":"Hollborn","year":"2007","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"496","DOI":"10.2353\/ajpath.2010.080642","article-title":"Crosstalk between vascular endothelial growth factor and matrix metalloproteinases in the induction of neovascularization in vivo","volume":"176","author":"Ebrahem","year":"2010","journal-title":"Am. J. Pathol."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"17","DOI":"10.1016\/j.preteyeres.2015.07.002","article-title":"Progress in corneal wound healing","volume":"49","author":"Ljubimov","year":"2015","journal-title":"Prog. Retin. Eye Res."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"511","DOI":"10.1006\/exer.2000.0979","article-title":"The epidermal growth factor receptor (EGFR): Role on corneal wound healing and homeostasis","volume":"72","author":"Nakamura","year":"2001","journal-title":"Exp. Eye Res."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"101","DOI":"10.1016\/j.smim.2009.01.004","article-title":"Plasmacytoid dendritic cells in the skin: To sense or not to sense nucleic acids","volume":"21","author":"Conrad","year":"2009","journal-title":"Semin. Immunol."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"2257","DOI":"10.1096\/fj.06-7918com","article-title":"Interdependence of lipoxin A4 and heme-oxygenase in counter-regulating inflammation during corneal wound healing","volume":"21","author":"Biteman","year":"2007","journal-title":"FASEB J."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"15267","DOI":"10.1074\/jbc.M410638200","article-title":"A role for the mouse 12\/15-lipoxygenase pathway in promoting epithelial wound healing and host defense","volume":"280","author":"Gronert","year":"2005","journal-title":"J. Biol. Chem."}],"container-title":["Biomolecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2218-273X\/13\/5\/831\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T19:34:19Z","timestamp":1760124859000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2218-273X\/13\/5\/831"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,5,13]]},"references-count":52,"journal-issue":{"issue":"5","published-online":{"date-parts":[[2023,5]]}},"alternative-id":["biom13050831"],"URL":"https:\/\/doi.org\/10.3390\/biom13050831","relation":{},"ISSN":["2218-273X"],"issn-type":[{"value":"2218-273X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,5,13]]}}}