Oxidative stress, autophagy and pyroptosis in the neovascularization of oxygen‑induced retinopathy in mice Erratum in /10.3892/mmr.2020.11753

Wang,Shuai; Ji,Li‑Yang; Li,Li; Li,Jing‑Min

doi:10.3892/mmr.2018.9759

Oxidative stress, autophagy and pyroptosis in the neovascularization of oxygen‑induced retinopathy in mice

Erratum in: /10.3892/mmr.2020.11753

Authors:
- Shuai Wang
- Li‑Yang Ji
- Li Li
- Jing‑Min Li
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Affiliations: Department of Ophthalmology, Second Affiliated Hospital of Dalian Medical University, Dalian, Liaoning 116023, P.R. China, Department of Ophthalmology, Zaozhuang Municipal Hospital, Zaozhuang, Shandong 277000, P.R. China
Published online on: December 13, 2018 https://doi.org/10.3892/mmr.2018.9759
Pages: 927-934
Copyright: © Wang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Retinal neovascularization (RNV) is a principal cause of visual impairment and blindness worldwide. The present study aimed to investigate how oxidative stress, autophagy and pyroptosis alter in RNV. The oxygen‑induced retinopathy (OIR) model was established in C57BL/6J mice by exposing them to a high concentration of oxygen. RNV was clearly visible in the fundus images and was qualitatively analyzed by counting the number of neovascular endothelial cell nuclei at postnatal day 17. Subsequently, the expression of vascular endothelial growth factor (VEGF)‑A and hypoxia‑inducible factor‑1α (HIF‑1α) at the protein level were measured. Furthermore, oxidative stress was examined using dihydroethidium (DHE) staining, and NADPH oxidase (NOX) 1 and 4 in the retinas were detected using reverse transcription‑quantitative polymerase chain reaction analysis. Additionally, immunostaining of microtubule associated protein 1 light chain 3α (LC3) was performed and the expression levels of the LC3, p62, autophagy protein (Atg)5, Atg7, Atg12, Beclin1, NOD‑like receptor family pyrin domain‑containing 3 (NLRP3), caspase‑1, interleukin (IL)‑1β, pro‑caspase‑1 and pro‑IL‑1β proteins were determined using western blotting in order to detect pyroptosis and autophagic flux. Autophagosomes were also detected using transmission electron microscopy. The results revealed that VEGF‑A and HIF‑1α protein expression levels, the DHE‑positive area, and NOX1 and NOX4 mRNA expression levels were significantly increased in the OIR mice. Furthermore, increased levels of NLRP3, caspase‑1, IL‑1β, pro‑caspase‑1 and pro‑IL‑1β proteins demonstrated that pyroptosis was activated. However, an accumulation of p62 and a reduction in the levels of LC3II/I and autophagosomes indicated that autophagic flux was compromised. Therefore, elevated levels of reactive oxygen species and pyroptosis along with attenuated autophagy were demonstrated in the OIR mice. The combination of oxidative stress, pyroptosis and impaired autophagy may serve an important role in the pathophysiology of RNV and may be a potential target to prevent RNV.

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1	Kim YW and Byzova TV: Oxidative stress in angiogenesis and vascular disease. Blood. 123:625–631. 2014. View Article : Google Scholar : PubMed/NCBI
2	Bhatt L, Groeger G, McDermott K and Cotter TG: Rod and cone photoreceptor cells produce ROS in response to stress in a live retinal explant system. Mol Vis. 16:283–293. 2010.PubMed/NCBI
3	Al-Shabrawey M, Bartoli M, El-Remessy AB, Platt DH, Matragoon S, Behzadian MA, Caldwell RW and Caldwell RB: Inhibition of NAD(P)H oxidase activity blocks vascular endothelial growth factor overexpression and neovascularization during ischemic retinopathy. Am J Pathol. 167:599–607. 2005. View Article : Google Scholar : PubMed/NCBI
4	Guo C, Yang M, Jing L, Wang J, Yu Y, Li Y, Duan J, Zhou X, Li Y and Sun Z: Amorphous silica nanoparticles trigger vascular endothelial cell injury through apoptosis and autophagy via reactive oxygen species-mediated MAPK/Bcl-2 and PI3K/Akt/mTOR signaling. Int J Nanomed. 11:5257–5276. 2016. View Article : Google Scholar
5	Adamis AP and Shima DT: The role of vascular endothelial growth factor in ocular health and disease. Retina. 25:111–118. 2005. View Article : Google Scholar : PubMed/NCBI
6	Zepeda AB, Pessoa A Jr, Castillo RL, Figueroa CA, Pulgar VM and Farías JG: Cellular and molecular mechanisms in the hypoxic tissue: Role of HIF-1 and ROS. Cell Biochem Funct. 31:451–459. 2013. View Article : Google Scholar : PubMed/NCBI
7	Saito S, Lin YC, Tsai MH, Lin CS, Murayama Y, Sato R and Yokoyama KK: Emerging roles of hypoxia-inducible factors and reactive oxygen species in cancer and pluripotent stem cells. Kaohsiung J Med Sci. 31:279–286. 2015. View Article : Google Scholar : PubMed/NCBI
8	Kietzmann T and Görlach A: Reactive oxygen species in the control of hypoxia-inducible factor-mediated gene expression. Semin Cell Dev Biol. 16:474–486. 2005. View Article : Google Scholar : PubMed/NCBI
9	Green DR, Galluzzi L and Kroemer G: Mitochondria and the autophagy-inflammation-cell death axis in organismal aging. Science. 333:1109–1112. 2011. View Article : Google Scholar : PubMed/NCBI
10	Vande Walle L and Lamkanfi M: Pyroptosis. Curr Biol. 26:R568–R572. 2016. View Article : Google Scholar : PubMed/NCBI
11	Man SM, Karki R and Kanneganti TD: Molecular mechanisms and functions of pyroptosis, inflammatory caspases and inflammasomes in infectious diseases. Immunol Rev. 277:61–75. 2017. View Article : Google Scholar : PubMed/NCBI
12	Li L, Tan J, Miao Y, Lei P and Zhang Q: ROS and autophagy: Interactions and molecular regulatory mechanisms. Cell Mol Neurobiol. 35:615–621. 2015. View Article : Google Scholar : PubMed/NCBI
13	Levonen AL, Hill BG, Kansanen E, Zhang J and Darley-Usmar VM: Redox regulation of antioxidants, autophagy, and the response to stress: Implications for electrophile therapeutics. Free Radic Biol Med. 71:196–207. 2014. View Article : Google Scholar : PubMed/NCBI
14	Lapaquette P, Guzzo J, Bretillon L and Bringer MA: Cellular and molecular connections between autophagy and inflammation. Mediators Inflamm. 2015:3984832015. View Article : Google Scholar : PubMed/NCBI
15	Blasiak J, Petrovski G, Veréb Z, Facskó A and Kaarniranta K: Oxidative stress, hypoxia, and autophagy in the neovascular processes of age-related macular degeneration. Biomed Res Int. 2014:7680262014. View Article : Google Scholar : PubMed/NCBI
16	Grossniklaus HE, Kang SJ and Berglin L: Animal models of choroidal and retinal neovascularization. Prog Retin Eye Res. 29:500–519. 2010. View Article : Google Scholar : PubMed/NCBI
17	Kilkenny C, Browne WJ, Cuthill IC, Emerson M and Altman DG: Improving bioscience research reporting: The ARRIVE guidelines for reporting animal research. PLoS Biol. 8:e10004122010. View Article : Google Scholar : PubMed/NCBI
18	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
19	Wilkinson-Berka JL, Deliyanti D, Rana I, Miller AG, Agrotis A, Armani R, Szyndralewiez C, Wingler K, Touyz RM, Cooper ME, et al: NADPH oxidase, NOX1, mediates vascular injury in ischemic retinopathy. Antioxid Redox Signal. 20:2726–2740. 2014. View Article : Google Scholar : PubMed/NCBI
20	Li J, Wang JJ, Yu Q, Chen K, Mahadev K and Zhang SX: Inhibition of reactive oxygen species by Lovastatin downregulates vascular endothelial growth factor expression and ameliorates blood-retinal barrier breakdown in db/db mice: Role of NADPH oxidase 4. Diabetes. 59:1528–1538. 2010. View Article : Google Scholar : PubMed/NCBI
21	Harris J: Autophagy and cytokines. Cytokine. 56:140–144. 2011. View Article : Google Scholar : PubMed/NCBI
22	Klionsky DJ, Abdelmohsen K, Abe A, Abedin MJ, Abeliovich H, Acevedo Arozena A, Adachi H, Adams CM, Adams PD, Adeli K, et al: Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition). Autophagy. 12:1–222. 2016. View Article : Google Scholar : PubMed/NCBI
23	Zin A and Gole GA: Retinopathy of prematurity-incidence today. Clin Perinatol. 40:185–200. 2013. View Article : Google Scholar : PubMed/NCBI
24	Gilbert C, Fielder A, Gordillo L, Quinn G, Semiglia R, Visintin P and Zin A: International NO-ROP Group: Characteristics of infants with severe retinopathy of prematurity in countries with low, moderate, and high levels of development: Implications for screening programs. Pediatrics. 115:e518–e525. 2005. View Article : Google Scholar : PubMed/NCBI
25	Cabral T, Mello LGM, Lima LH, Polido J, Regatieri CV, Belfort R Jr and Mahajan VB: Retinal and choroidal angiogenesis: A review of new targets. Int J Retina Vitreous. 3:312017. View Article : Google Scholar : PubMed/NCBI
26	Cervia D, Catalani E, Dal Monte M and Casini G: Vascular endothelial growth factor in the ischemic retina and its regulation by somatostatin. J Neurochem. 120:818–829. 2012. View Article : Google Scholar : PubMed/NCBI
27	Finkel T and Holbrook NJ: Oxidants, oxidative stress and the biology of ageing. Nature. 408:239–247. 2000. View Article : Google Scholar : PubMed/NCBI
28	Gu Q, He Y, Ji J, Yao Y, Shen W, Luo J, Zhu W, Cao H, Geng Y, Xu J, et al: Hypoxia-inducible factor 1α (HIF-1α) and reactive oxygen species (ROS) mediates radiation-induced invasiveness through the SDF-1α/CXCR4 pathway in non-small cell lung carcinoma cells. Oncotarget. 6:10893–10907. 2015. View Article : Google Scholar : PubMed/NCBI
29	Andrei C, Dazzi C, Lotti L, Torrisi MR, Chimini G and Rubartelli A: The secretory route of the leaderless protein interleukin 1beta involves exocytosis of endolysosome-related vesicles. Mol Biol Cell. 10:1463–1475. 1999. View Article : Google Scholar : PubMed/NCBI
30	Miao EA, Rajan JV and Aderem A: Caspase-1-induced pyroptotic cell death. Immunol Rev. 243:206–214. 2011. View Article : Google Scholar : PubMed/NCBI
31	Zhou R, Tardivel A, Thorens B, Choi I and Tschopp J: Thioredoxin-interacting protein links oxidative stress to inflammasome activation. Nat Immunol. 11:136–140. 2009. View Article : Google Scholar : PubMed/NCBI
32	Wu D, Han R, Deng S, Liu T, Zhang T, Xie H and Xu Y: Protective effects of flagellin A N/C against radiation-induced NLR pyrin domain containing 3 inflammasome-dependent pyroptosis in intestinal cells. Int J Radiat Oncol Biol Phys. 101:107–117. 2018. View Article : Google Scholar : PubMed/NCBI
33	Kim JY, Paton JC, Briles DE, Rhee DK and Pyo S: Streptococcus pneumoniae induces pyroptosis through the regulation of autophagy in murine microglia. Oncotarget. 6:44161–44178. 2015. View Article : Google Scholar : PubMed/NCBI
34	Szumiel I: Autophagy, reactive oxygen species and the fate of mammalian cells. Free Radic Res. 45:253–265. 2011. View Article : Google Scholar : PubMed/NCBI
35	Filomeni G, Desideri E, Cardaci S, Rotilio G and Ciriolo MR: Under the ROS…thiol network is the principal suspect for autophagy commitment. Autophagy. 6:999–1005. 2014. View Article : Google Scholar
36	Mitter SK, Song C, Qi X, Mao H, Rao H, Akin D, Lewin A, Grant M, Dunn W Jr, Ding J, et al: Dysregulated autophagy in the RPE is associated with increased susceptibility to oxidative stress and AMD. Autophagy. 10:1989–2005. 2014. View Article : Google Scholar : PubMed/NCBI
37	Kaarniranta K, Sinha D, Blasiak J, Kauppinen A, Veréb Z, Salminen A, Boulton ME and Petrovski G: Autophagy and heterophagy dysregulation leads to retinal pigment epithelium dysfunction and development of age-related macular degeneration. Autophagy. 9:973–984. 2013. View Article : Google Scholar : PubMed/NCBI
38	Lopes de Faria JM, Duarte DA, Montemurro C, Papadimitriou A, Consonni SR and Lopes de Faria JB: Defective autophagy in diabetic retinopathy. Invest Ophthalmol Vis Sci. 57:4356–4366. 2016. View Article : Google Scholar : PubMed/NCBI
39	Liu J, Copland DA, Theodoropoulou S, Chiu HA, Barba MD, Mak KW, Mack M, Nicholson LB and Dick AD: Impairing autophagy in retinal pigment epithelium leads to inflammasome activation and enhanced macrophage-mediated angiogenesis. Sci Rep. 6:206392016. View Article : Google Scholar : PubMed/NCBI
40	Wang Y, Hanus J, Abu-Asab M, Shen D, Ogilvy A, Ou J, Chu XK, Shi G, Li W, Wang S and Chan CC: NLRP3 upregulation in retinal pigment epithelium in age-related macular degeneration. Int J Mol Sci. 17(pii): E732016. View Article : Google Scholar : PubMed/NCBI
41	Sen CK, Khanna S, Babior BM, Hunt TK, Ellison EC and Roy S: Oxidant-induced vascular endothelial growth factor expression in human keratinocytes and cutaneous wound healing. J Biol Chem. 277:33284–33290. 2002. View Article : Google Scholar : PubMed/NCBI
42	Schreml S, Szeimies RM, Prantl L, Karrer S, Landthaler M and Babilas P: Oxygen in acute and chronic wound healing. Br J Dermatol. 163:257–268. 2010. View Article : Google Scholar : PubMed/NCBI
43	Chung AS and Ferrara N: Developmental and Pathological Angiogenesis. Annu Rev Cell Dev Biol. 27:563–584. 2011. View Article : Google Scholar : PubMed/NCBI
44	Moreno ML, Mérida S, Bosch-Morell F, Miranda M and Villar VM: Autophagy dysfunction and oxidative stress, two related mechanisms implicated in retinitis pigmentosa. Front Physiol. 9:10082018. View Article : Google Scholar : PubMed/NCBI
45	Sureshbabu A, Ryter SW and Choi ME: Oxidative stress and autophagy: Crucial modulators of kidney injury. Redox Biol. 4:208–214. 2015. View Article : Google Scholar : PubMed/NCBI