Peptain-1 Blocks Ischemia/reperfusion-induced Retinal Capillary Degeneration in Mice

Overview

Journal Front Cell Neurosci

Specialty Cell Biology

Date 2024 Aug 16

PMID 39149168

Authors

Mi-Hyun Nam

Armaan Dhillon

Rooban B Nahomi

Noelle L Carrillo

Clarinda S Hougen

Ram H Nagaraj

Affiliations

Soon will be listed here.

Abstract

Introduction: Neurovascular degeneration results in vascular dysfunction, leakage, ischemia, and structural changes that can lead to significant visual impairment. We previously showed the protective effects of peptain-1, a 20 amino acid peptide derived from the αB-crystallin core domain, on retinal ganglion cells in two animal models of glaucoma. Here, we evaluated the ability of peptain-1 to block apoptosis of human retinal endothelial cells (HRECs) and retinal capillary degeneration in mice subjected to retinal ischemia/reperfusion (I/R) injury.

Methods: HRECs were treated with either peptain-1 or scrambled peptides (200 μg/mL) for 3 h and a combination of proinflammatory cytokines (IFN-γ 20 ng/mL + TNF-α 20 ng/mL+ IL-1β 20 ng/mL) for additional 48 h. Apoptosis was measured with cleaved caspase-3 formation via western blot, and by TUNEL assay. C57BL/6J mice (12 weeks old) were subjected to I/R injury by elevating the intraocular pressure to 120 mmHg for 60 min, followed by reperfusion. Peptain-1 or scrambled peptide (0.5 μg) was intravitreally injected immediately after I/R injury and 7 days later. One microliter of PBS was injected as vehicle control, and animals were euthanized on day 14 post-I/R injury. Retinal capillary degeneration was assessed after enzyme digestion followed by periodic acid-Schiff staining.

Results: Our data showed that peptain-1 entered HRECs and blocked proinflammatory cytokine-mediated apoptosis. Intravitreally administered peptain-1 was distributed throughout the retinal vessels after 4 h. I/R injury caused retinal capillary degeneration. Unlike scrambled peptide, peptain-1 protected capillaries against I/R injury. Additionally, peptain-1 inhibited microglial activation and reduced proinflammatory cytokine levels in the retina following I/R injury.

Discussion: Our study suggests that peptain-1 could be used as a therapeutic agent to prevent capillary degeneration and neuroinflammation in retinal ischemia.

References

Hammes H, Lin J, Renner O, Shani M, Lundqvist A, Betsholtz C . Pericytes and the pathogenesis of diabetic retinopathy. Diabetes. 2002; 51(10):3107-12. DOI: 10.2337/diabetes.51.10.3107. View

Watson E, Grant Z, Coultas L . Endothelial cell apoptosis in angiogenesis and vessel regression. Cell Mol Life Sci. 2017; 74(24):4387-4403. PMC: 11107683. DOI: 10.1007/s00018-017-2577-y. View

Nagaraj R, Nahomi R, Mueller N, Raghavan C, Ammar D, Petrash J . Therapeutic potential of α-crystallin. Biochim Biophys Acta. 2015; 1860(1 Pt B):252-7. PMC: 4591086. DOI: 10.1016/j.bbagen.2015.03.012. View

Nahomi R, Palmer A, Green K, Fort P, Nagaraj R . Pro-inflammatory cytokines downregulate Hsp27 and cause apoptosis of human retinal capillary endothelial cells. Biochim Biophys Acta. 2013; 1842(2):164-74. PMC: 3905326. DOI: 10.1016/j.bbadis.2013.11.011. View

Osborne N, Casson R, Wood J, Chidlow G, Graham M, Melena J . Retinal ischemia: mechanisms of damage and potential therapeutic strategies. Prog Retin Eye Res. 2004; 23(1):91-147. DOI: 10.1016/j.preteyeres.2003.12.001. View

Heise E, Fort P . Impact of diabetes on alpha-crystallins and other heat shock proteins in the eye. J Ocul Biol Dis Infor. 2012; 4(1-2):62-9. PMC: 3342406. DOI: 10.1007/s12177-011-9073-7. View

Xu G, Liu B, Sun Y, Du Y, Snetselaar L, Hu F . Prevalence of diagnosed type 1 and type 2 diabetes among US adults in 2016 and 2017: population based study. BMJ. 2018; 362:k1497. PMC: 6122253. DOI: 10.1136/bmj.k1497. View

Kannan R, Sreekumar P, Hinton D . Novel roles for α-crystallins in retinal function and disease. Prog Retin Eye Res. 2012; 31(6):576-604. PMC: 3472046. DOI: 10.1016/j.preteyeres.2012.06.001. View

Andley U, Song Z, Wawrousek E, Fleming T, Bassnett S . Differential protective activity of alpha A- and alphaB-crystallin in lens epithelial cells. J Biol Chem. 2000; 275(47):36823-31. DOI: 10.1074/jbc.M004233200. View

10.

Stitt A, Curtis T, Chen M, Medina R, Mckay G, Jenkins A . The progress in understanding and treatment of diabetic retinopathy. Prog Retin Eye Res. 2015; 51:156-86. DOI: 10.1016/j.preteyeres.2015.08.001. View

11.

Nam M, Smith A, Pantcheva M, Park K, Brzezinski J, Galligan J . Aspirin inhibits TGFβ2-induced epithelial to mesenchymal transition of lens epithelial cells: selective acetylation of K56 and K122 in histone H3. Biochem J. 2019; 477(1):75-97. PMC: 8259308. DOI: 10.1042/BCJ20190540. View

12.

Fu X, Gens J, Glazier J, Burns S, Gast T . Progression of Diabetic Capillary Occlusion: A Model. PLoS Comput Biol. 2016; 12(6):e1004932. PMC: 4907516. DOI: 10.1371/journal.pcbi.1004932. View

13.

Chen G, Yan F, Wei W, Wang F, Wang Z, Nie J . CD38 deficiency protects the retina from ischaemia/reperfusion injury partly via suppression of TLR4/MyD88/NF-κB signalling. Exp Eye Res. 2022; 219:109058. DOI: 10.1016/j.exer.2022.109058. View

14.

Shin E, Sorenson C, Sheibani N . Diabetes and retinal vascular dysfunction. J Ophthalmic Vis Res. 2015; 9(3):362-73. PMC: 4307665. DOI: 10.4103/2008-322X.143378. View

15.

Joussen A, Doehmen S, Le M, Koizumi K, Radetzky S, Krohne T . TNF-alpha mediated apoptosis plays an important role in the development of early diabetic retinopathy and long-term histopathological alterations. Mol Vis. 2009; 15:1418-28. PMC: 2716944. View

16.

Ucgun N, Zeki-Fikret C, Yildirim Z . Inflammation and diabetic retinopathy. Mol Vis. 2020; 26:718-721. PMC: 7655973. View

17.

Khan M, Hashim M, King J, Govender R, Mustafa H, Al Kaabi J . Epidemiology of Type 2 Diabetes - Global Burden of Disease and Forecasted Trends. J Epidemiol Glob Health. 2020; 10(1):107-111. PMC: 7310804. DOI: 10.2991/jegh.k.191028.001. View

18.

Homme R, Singh M, Majumder A, George A, Nair K, Sandhu H . Remodeling of Retinal Architecture in Diabetic Retinopathy: Disruption of Ocular Physiology and Visual Functions by Inflammatory Gene Products and Pyroptosis. Front Physiol. 2018; 9:1268. PMC: 6134046. DOI: 10.3389/fphys.2018.01268. View

19.

Nahomi R, Huang R, Nandi S, Wang B, Padmanabha S, Santhoshkumar P . Acetylation of lysine 92 improves the chaperone and anti-apoptotic activities of human αB-crystallin. Biochemistry. 2013; 52(45):8126-38. PMC: 3889837. DOI: 10.1021/bi400638s. View

20.

Zhao Y, Singh R . The role of anti-vascular endothelial growth factor (anti-VEGF) in the management of proliferative diabetic retinopathy. Drugs Context. 2018; 7:212532. PMC: 6113746. DOI: 10.7573/dic.212532. View