Abstract
Dry eye disease (DED) is a prevalent ocular disorder with a multifactorial etiology. The pre-angiogenic and pre-inflammatory milieu of the ocular surface plays a critical role in its pathogenesis. DZ2002 is a reversible type III S-adenosyl-L-homocysteine hydrolase (SAHH) inhibitor, which has shown excellent anti-inflammatory and immunosuppressive activities in vivo and in vitro. In this study, we evaluated the therapeutic potential of DZ2002 in rodent models of DED. SCOP-induced dry eye models were established in female rats and mice, while BAC-induced dry eye model was established in female rats. DZ2002 was administered as eye drops (0.25%, 1%) four times daily (20 μL per eye) for 7 or 14 consecutive days. We showed that topical application of DZ2002 concentration-dependently reduced corneal neovascularization and corneal opacity, as well as alleviated conjunctival irritation in both DED models. Furthermore, we observed that DZ2002 treatment decreased the expression of genes associated with angiogenesis and the levels of inflammation in the cornea and conjunctiva. Moreover, DZ2002 treatment in the BAC-induced DED model abolished the activation of the STAT3-PI3K-Akt-NF-κB pathways in corneal tissues. We also found that DZ2002 significantly inhibited the proliferation, migration, and tube formation of human umbilical endothelial cells (HUVECs) while downregulating the activation of the STAT3-PI3K-Akt-NF-κB pathway. These results suggest that DZ2002 exerts a therapeutic effect on corneal angiogenesis in DED, potentially by preventing the upregulation of the STAT3-PI3K-Akt-NF-κB pathways. Collectively, DZ2002 is a promising candidate for ophthalmic therapy, particularly in treating DED.
Similar content being viewed by others
References
Craig JP, Nichols KK, Akpek EK, Caffery B, Dua HS, Joo CK, et al. TFOS DEWS II definition and classification report. Ocul Surf. 2017;15:276–83.
Di Zazzo A, Gaudenzi D, Yin J, Coassin M, Fernandes M, Dana R, et al. Corneal angiogenic privilege and its failure. Exp Eye Res. 2021;204:108457.
Lyu N, Zhao Y, **ang J, Fan X, Huang C, Sun X, et al. Inhibiting corneal neovascularization by sustainably releasing anti-VEGF and anti-inflammation drugs from silica-thermogel nanohybrids. Mater Sci Eng C Mater Biol Appl. 2021;128:112274.
Qazi Y, Wong G, Monson B, Stringham J, Ambati BK. Corneal transparency: genesis, maintenance and dysfunction. Brain Res Bull. 2010;81:198–210.
Coster DJ, Williams KA. The impact of corneal allograft rejection on the long-term outcome of corneal transplantation. Am J Ophthalmol. 2005;140:1112–22.
Hu J, Lin S, Huang JJ, Cheung PCK. Mechanistic study of the in vitro and in vivo inhibitory effects of protocatechuic acid and syringic acid on vegf-induced angiogenesis. J Agric Food Chem. 2018;66:6742–51.
Lennikov A, Mirabelli P, Mukwaya A, Schaupper M, Thangavelu M, Lachota M, et al. Selective IKK2 inhibitor IMD0354 disrupts NF-kappaB signaling to suppress corneal inflammation and angiogenesis. Angiogenesis. 2018;21:267–85.
Mirabelli P, Peebo BB, Xeroudaki M, Koulikovska M, Lagali N. Early effects of dexamethasone and anti-VEGF therapy in an inflammatory corneal neovascularization model. Exp Eye Res. 2014;125:118–27.
Ponnaluri VKC, Esteve PO, Ruse CI, Pradhan S. S-adenosylhomocysteine hydrolase participates in DNA methylation inheritance. J Mol Biol. 2018;430:2051–65.
He SJ, Lin ZM, Wu YW, Bai BX, Yang XQ, He PL, et al. Therapeutic effects of DZ2002, a reversible SAHH inhibitor, on lupus-prone NZBxNZW F1 mice via interference with TLR-mediated APC response. Acta Pharmacol Sin. 2014;35:219–29.
He S, Liu X, Lin Z, Liu Y, Gu L, Zhou H, et al. Reversible SAHH inhibitor protects against glomerulonephritis in lupus-prone mice by downregulating renal alpha-actinin-4 expression and stabilizing integrin-cytoskeleton linkage. Arthritis Res Ther. 2019;21:40.
Lee SJ, Im ST, Wu J, Cho CS, Jo DH, Chen Y, et al. Corneal lymphangiogenesis in dry eye disease is regulated by substance P/neurokinin-1 receptor system through controlling expression of vascular endothelial growth factor receptor 3. Ocul Surf. 2021;22:72–79.
Yang FM, Fan D, Yang XQ, Zhu FH, Shao MJ, Li Q, et al. The artemisinin analog SM934 alleviates dry eye disease in rodent models by regulating TLR4/NF-kappaB/NLRP3 signaling. Acta Pharmacol Sin. 2021;42:593–603.
Pauly A, Brignole-Baudouin F, Labbe A, Liang H, Warnet JM, Baudouin C. New tools for the evaluation of toxic ocular surface changes in the rat. Invest Ophthalmol Vis Sci. 2007;48:5473–83.
Nakatani H, Gomes P, Bradford R, Guo Q, Safyan E, Hollander DA. Alcaftadine 0.25% versus Olopatadine 0.1% in preventing cedar pollen allergic conjunctivitis in Japan: a randomized study. Ocul Immunol Inflamm. 2019;27:622–31.
Na YJ, Choi KJ, Park SB, Sung HR, Jung WH, Kim HY, et al. Protective effects of carbenoxolone, an 11beta-HSD1 inhibitor, against chemical induced dry eye syndrome. Apoptosis. 2017;22:1441–53.
Lin ZM, Liu YT, Huang YT, Yang XQ, Zhu FH, Tang W, et al. Anti-nociceptive, anti-inflammatory and anti-arthritic activities of pregnane glycosides from the root bark of periploca sepium bunge. J Ethnopharmacol. 2021;265:113345.
He S, Ding H, Chen L, Shen Y, Liu Y, Zhu F, et al. Repression of interferon regulatory factor-4 (IRF4) hyperactivation restricts murine lupus. Signal Transduct Target Ther. 2023;8:188.
Lin ZM, Ma M, Li H, Qi Q, Liu YT, Yan YX, et al. Topical administration of reversible SAHH inhibitor ameliorates imiquimod-induced psoriasis-like skin lesions in mice via suppression of TNF-alpha/IFN-gamma-induced inflammatory response in keratinocytes and T cell-derived IL-17. Pharmacol Res. 2018;129:443–52.
Chin HK, Horng CT, Liu YS, Lu CC, Su CY, Chen PS, et al. Kaempferol inhibits angiogenic ability by targeting VEGF receptor-2 and downregulating the PI3K/AKT, MEK and ERK pathways in VEGF-stimulated human umbilical vein endothelial cells. Oncol Rep. 2018;39:2351–7.
Chen L, Lin Z, Liu Y, Cao S, Huang Y, Yang X, et al. DZ2002 alleviates psoriasis-like skin lesions via differentially regulating methylation of GATA3 and LCN2 promoters. Int Immunopharmacol. 2021;91:107334.
Yang Q, Zhang Y, Liu X, Wang N, Song Z, Wu K. A Comparison of the effects of benzalkonium chloride on ocular surfaces between C57BL/6 and BALB/c mice. Int J Mol Sci. 2017;18:509.
Chen W, Li Z, Hu J, Zhang Z, Chen L, Chen Y, et al. Corneal alternations induced by topical application of benzalkonium chloride in rabbit. PLoS One. 2011;6:e26103.
Lin Z, Liu X, Zhou T, Wang Y, Bai L, He H, et al. A mouse dry eye model induced by topical administration of benzalkonium chloride. Mol Vis. 2011;17:257–64.
Shanmugham V, Subban R. Capsanthin from capsicum annum fruits exerts anti-glaucoma, antioxidant, anti-inflammatory activity, and corneal pro-inflammatory cytokine gene expression in a benzalkonium chloride-induced rat dry eye model. J Food Biochem. 2022;46:e14352.
Carpena-Torres C, Pintor J, Perez de Lara MJ, Huete-Toral F, Crooke A, Pastrana C, et al. Optimization of a rabbit dry eye model induced by topical instillation of benzalkonium chloride. J Ophthalmol. 2020;2020:7204951.
Bock F, Maruyama K, Regenfuss B, Hos D, Steven P, Heindl LM, et al. Novel anti(lymph)angiogenic treatment strategies for corneal and ocular surface diseases. Prog Retin Eye Res. 2013;34:89–124.
Ji YW, Lee JL, Kang HG, Gu N, Byun H, Yeo A, et al. Corneal lymphangiogenesis facilitates ocular surface inflammation and cell trafficking in dry eye disease. Ocul Surf. 2018;16:306–13.
Liu C, He L, Wang J, Wang Q, Sun C, Li Y, et al. Anti-angiogenic effect of Shikonin in rheumatoid arthritis by downregulating PI3K/AKT and MAPKs signaling pathways. J Ethnopharmacol. 2020;260:113039.
Patnam M, Dommaraju SR, Masood F, Herbst P, Chang JH, Hu WY, et al. Lymphangiogenesis guidance mechanisms and therapeutic implications in pathological states of the cornea. Cells. 2023;12:319.
Yang T, Wang X, Guo L, Zheng F, Meng C, Zheng Y, et al. Daphnetin inhibits corneal inflammation and neovascularization on a mouse model of corneal alkali burn. Int Immunopharmacol. 2022;103:108434.
Eccles SA. Parallels in invasion and angiogenesis provide pivotal points for therapeutic intervention. Int J Dev Biol. 2004;48:583–98.
Yang W, Yang Y, Wan S, Xu Y, Li J, Zhang L, et al. Exploring the mechanism of the miRNA-145/Paxillin axis in cell metabolism during VEGF-A-induced corneal angiogenesis. Invest Ophthalmol Vis Sci. 2021;62:25.
Meng N, **e HX, Hou JR, Chen YB, Wu MJ, Guo YW, et al. Design and semisynthesis of oleanolic acid derivatives as VEGF inhibitors: Inhibition of VEGF-induced proliferation, angiogenesis, and VEGFR2 activation in HUVECs. Chin J Nat Med. 2022;20:229–40.
Ouyang B, **e Y, Zhang C, Deng C, Lv L, Yao J, et al. Extracellular vesicles from human urine-derived stem cells ameliorate erectile dysfunction in a diabetic rat model by delivering proangiogenic microRNA. Sex Med. 2019;7:241–50.
Rezzola S, Belleri M, Gariano G, Ribatti D, Costagliola C, Semeraro F, et al. In vitro and ex vivo retina angiogenesis assays. Angiogenesis. 2014;17:429–42.
Zhong W, Montana M, Santosa SM, Isjwara ID, Huang YH, Han KY, et al. Angiogenesis and lymphangiogenesis in corneal transplantation—a review. Surv Ophthalmol. 2018;63:453–79.
Kim M, Lee C, Payne R, Yue BY, Chang JH, Ying H. Angiogenesis in glaucoma filtration surgery and neovascular glaucoma: a review. Surv Ophthalmol. 2015;60:524–35.
Apte RS, Chen DS, Ferrara N. VEGF in signaling and disease: beyond discovery and development. Cell. 2019;176:1248–64.
Nicholas MP, Mysore N. Corneal neovascularization. Exp Eye Res. 2021;202:108363.
Droy-Lefaix MT, Bueno L, Caron P, Belot E, Roche O. Ocular inflammation and corneal permeability alteration by benzalkonium chloride in rats: a protective effect of a myosin light chain kinase inhibitor. Invest Ophthalmol Vis Sci. 2013;54:2705–10.
Chen Y, Chauhan SK, Lee HS, Stevenson W, Schaumburg CS, Sadrai Z, et al. Effect of desiccating environmental stress versus systemic muscarinic AChR blockade on dry eye immunopathogenesis. Invest Ophthalmol Vis Sci. 2013;54:2457–64.
Baudouin C, Rolando M, Benitez Del Castillo JM, Messmer EM, Figueiredo FC, Irkec M, et al. Reconsidering the central role of mucins in dry eye and ocular surface diseases. Prog Retin Eye Res. 2019;71:68–87.
Dartt DA, Masli S. Conjunctival epithelial and goblet cell function in chronic inflammation and ocular allergic inflammation. Curr Opin Allergy Clin Immunol. 2014;14:464–70.
Contreras-Ruiz L, Ghosh-Mitra A, Shatos MA, Dartt DA, Masli S. Modulation of conjunctival goblet cell function by inflammatory cytokines. Mediators Inflamm. 2013;2013:636812.
Stephens DN, McNamara NA. Altered mucin and glycoprotein expression in dry eye disease. Optom Vis Sci. 2015;92:931–8.
Zhang M, Tombran-Tink J, Yang S, Zhang X, Li X, Barnstable CJ. PEDF is an endogenous inhibitor of VEGF-R2 angiogenesis signaling in endothelial cells. Exp Eye Res. 2021;213:108828.
Cho HD, Kim JH, Park JK, Hong SM, Kim DH, Seo KI. Kochia scoparia seed extract suppresses VEGF-induced angiogenesis via modulating VEGF receptor 2 and PI3K/AKT/mTOR pathways. Pharm Biol. 2019;57:684–93.
Cho HD, Moon KD, Park KH, Lee YS, Seo KI. Effects of auriculasin on vascular endothelial growth factor (VEGF)-induced angiogenesis via regulation of VEGF receptor 2 signaling pathways in vitro and in vivo. Food Chem Toxicol. 2018;121:612–21.
Cho HD, Lee KW, Won YS, Kim JH, Seo KI. Cultivated Orostachys japonicus extract inhibits VEGF-induced angiogenesis via regulation of VEGFR2 signaling pathway in vitro and in vivo. J Ethnopharmacol. 2020;256:112664.
Sethi G, Sung B, Aggarwal BB. TNF: a master switch for inflammation to cancer. Front Biosci. 2008;13:5094–107.
Hadrian K, Willenborg S, Bock F, Cursiefen C, Eming SA, Hos D. Macrophage-mediated tissue vascularization: similarities and differences between cornea and skin. Front Immunol. 2021;12:667830.
Spiller KL, Anfang RR, Spiller KJ, Ng J, Nakazawa KR, Daulton JW, et al. The role of macrophage phenotype in vascularization of tissue engineering scaffolds. Biomaterials. 2014;35:4477–88.
Hall KL, Volk-Draper LD, Flister MJ, Ran S. New model of macrophage acquisition of the lymphatic endothelial phenotype. PLoS One. 2012;7:e31794.
Corliss BA, Azimi MS, Munson JM, Peirce SM, Murfee WL. Macrophages: an inflammatory link between angiogenesis and lymphangiogenesis. Microcirculation. 2016;23:95–121.
Acknowledgements
We are thankful for financial support from the National Natural Science Foundation of China (81871240), Shanghai Municipal Science and Technology Major Project and the CAS “Light of West China” Program and CAS Interdisciplinary Innovation Team.
Author information
Authors and Affiliations
Contributions
ZML, SJH, and JPZ contributed to the conception and design of this study; CMW, JWM, XQY, and CCX completed the main part of the experiment; JYY, JWM, JZZ, MX, YFH, XT, DL, FHZ, and XYX participated in the in vivo and in vitro experiment; MNC participated in data statistic analysis; ZML, SJH, and JPZ contributed to drafting the manuscript and revising critically for important intellectual content.
Corresponding authors
Ethics declarations
Competing interests
The authors declare no competing interests.
Rights and permissions
Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Wu, Cm., Mao, Jw., Zhu, Jz. et al. DZ2002 alleviates corneal angiogenesis and inflammation in rodent models of dry eye disease via regulating STAT3-PI3K-Akt-NF-κB pathway. Acta Pharmacol Sin 45, 166–179 (2024). https://doi.org/10.1038/s41401-023-01146-y
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/s41401-023-01146-y
- Springer Nature Singapore Pte Ltd.