Effects of exosomes derived from activated corneal stromal keratocytes on the inflammation, proliferation, neuroprotection and epithelial-mesenchymal transition in retinal pigment epithelium cells

IF 5.2 2区医学 Q1 MEDICINE, RESEARCH & EXPERIMENTAL

Life sciences Pub Date : 2025-03-31 DOI:10.1016/j.lfs.2025.123592

Fatemeh Sanie-Jahromi , Niloofar Sadeghi , Zahra Moayedfard , Zahra Gharegezloo , Mahmood Nejabat , M. Hossein Nowroozzadeh

{"title":"Effects of exosomes derived from activated corneal stromal keratocytes on the inflammation, proliferation, neuroprotection and epithelial-mesenchymal transition in retinal pigment epithelium cells","authors":"Fatemeh Sanie-Jahromi , Niloofar Sadeghi , Zahra Moayedfard , Zahra Gharegezloo , Mahmood Nejabat , M. Hossein Nowroozzadeh","doi":"10.1016/j.lfs.2025.123592","DOIUrl":null,"url":null,"abstract":"<div><h3>Aims</h3><div>This study investigated the effects of activated keratocyte-derived exosomes (aKExo) on retinal pigment epithelial (RPE) cells <em>in-vitro</em>, focusing on cell viability, inflammatory cytokine expression, and neuroprotective properties.</div></div><div><h3>Materials and methods</h3><div>Keratocytes were cultured, and exosomes were extracted and characterized using transmission electron microscopy (TEM), scanning electron microscopy (SEM), flow cytometry, and dynamic light scattering (DLS). RPE cells, isolated from a human donor, were confirmed <em>via</em> RPE65 expression. aKExo effects on RPE cells were assessed using MTT assay at concentrations from 10<sup>−1</sup> (35 μg/mL) to 10<sup>−5</sup> (3.5 × 10<sup>−3</sup> μg/mL). The optimal aKExo concentration (10<sup>−5</sup>) enhanced cell viability and exhibited the highest proliferative potential compared to the control group, making it the optimal dose for subsequent experiments including gene expression analysis, and ELISA.</div></div><div><h3>Key findings</h3><div>aKExo downregulated IL-6 mRNA (0.70 ± 0.06, <em>p</em> = 0.0009) and marginally reduced TGF-β mRNA (0.75 ± 0.16, <em>p</em> = 0.0575). ELISA confirmed a reduction in IL-6 (31.33 ± 5.77 pg/mL <em>vs.</em> 50.22 ± 13.47 pg/mL, <em>p</em> = 0.0894) and TGF-β (8.91 ± 0.16 pg/mL <em>vs.</em> 11.39 ± 1.49 pg/mL, <em>p</em> = 0.0460). No significant changes were observed for IL-1β expression or other epithelial-mesenchymal transition (EMT)-related genes (α-SMA, ZEB-1, β-catenin). Neuroprotective genes NGF (4.34 ± 1.05, <em>p</em> = 0.0053) and CD90 (1.55 ± 0.25, <em>p</em> = 0.0184) were significantly upregulated, while VEGF-A was elevated (1.65 ± 0.15, <em>p</em> = 0.0018).</div></div><div><h3>Significance</h3><div>These findings highlight aKExo's immunomodulatory, neuroprotective, and anti-EMT effects, suggesting potential therapeutic applications for retinal disorders, while noting that VEGF-A upregulation requires further investigation.</div></div>","PeriodicalId":18122,"journal":{"name":"Life sciences","volume":"371 ","pages":"Article 123592"},"PeriodicalIF":5.2000,"publicationDate":"2025-03-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Life sciences","FirstCategoryId":"3","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S0024320525002267","RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"MEDICINE, RESEARCH & EXPERIMENTAL","Score":null,"Total":0}

引用次数: 0

Abstract

Aims

This study investigated the effects of activated keratocyte-derived exosomes (aKExo) on retinal pigment epithelial (RPE) cells in-vitro, focusing on cell viability, inflammatory cytokine expression, and neuroprotective properties.

Materials and methods

Keratocytes were cultured, and exosomes were extracted and characterized using transmission electron microscopy (TEM), scanning electron microscopy (SEM), flow cytometry, and dynamic light scattering (DLS). RPE cells, isolated from a human donor, were confirmed via RPE65 expression. aKExo effects on RPE cells were assessed using MTT assay at concentrations from 10⁻¹ (35 μg/mL) to 10⁻⁵ (3.5 × 10⁻³ μg/mL). The optimal aKExo concentration (10⁻⁵) enhanced cell viability and exhibited the highest proliferative potential compared to the control group, making it the optimal dose for subsequent experiments including gene expression analysis, and ELISA.

Key findings

aKExo downregulated IL-6 mRNA (0.70 ± 0.06, p = 0.0009) and marginally reduced TGF-β mRNA (0.75 ± 0.16, p = 0.0575). ELISA confirmed a reduction in IL-6 (31.33 ± 5.77 pg/mL vs. 50.22 ± 13.47 pg/mL, p = 0.0894) and TGF-β (8.91 ± 0.16 pg/mL vs. 11.39 ± 1.49 pg/mL, p = 0.0460). No significant changes were observed for IL-1β expression or other epithelial-mesenchymal transition (EMT)-related genes (α-SMA, ZEB-1, β-catenin). Neuroprotective genes NGF (4.34 ± 1.05, p = 0.0053) and CD90 (1.55 ± 0.25, p = 0.0184) were significantly upregulated, while VEGF-A was elevated (1.65 ± 0.15, p = 0.0018).

Significance

These findings highlight aKExo's immunomodulatory, neuroprotective, and anti-EMT effects, suggesting potential therapeutic applications for retinal disorders, while noting that VEGF-A upregulation requires further investigation.

查看原文本刊更多论文

活化的角膜基质角质细胞衍生的外泌体对视网膜色素上皮细胞炎症、增殖、神经保护和上皮-间质转化的影响

本研究探讨活化的角化细胞衍生外泌体（aKExo）对体外视网膜色素上皮（RPE）细胞的影响，重点关注细胞活力、炎症细胞因子表达和神经保护特性。材料和方法培养角质细胞，提取外泌体，并利用透射电镜（TEM）、扫描电镜（SEM）、流式细胞术和动态光散射（DLS）对其进行表征。从人供体中分离的RPE细胞通过表达RPE65得到证实。在浓度为10−1 （35 μg/mL）至10−5 （3.5 × 10−3 μg/mL）时，采用MTT法评估aKExo对RPE细胞的影响。与对照组相比，最佳aKExo浓度（10−5）提高了细胞活力，并表现出最高的增殖潜力，使其成为后续实验（包括基因表达分析和ELISA）的最佳剂量。关键发现sakexo下调IL-6 mRNA(0.70±0.06,p = 0.0009)，轻度下调TGF-β mRNA（0.75±0.16,p = 0.0575）。ELISA证实降低il - 6(31.33±5.77 pg / mL和50.22±13.47 pg / mL, p = 0.0894)和TGF -β(8.91±0.16 pg / mL和11.39±1.49 pg / mL, p = 0.0460)。IL-1β及其他上皮-间质转化（EMT）相关基因（α-SMA、ZEB-1、β-catenin）的表达未见显著变化。神经保护基因NGF（4.34±1.05,p = 0.0053）和CD90（1.55±0.25,p = 0.0184）显著上调，VEGF-A（1.65±0.15,p = 0.0018）显著升高。这些发现突出了aKExo的免疫调节、神经保护和抗emt作用，提示了视网膜疾病的潜在治疗应用，同时注意到VEGF-A上调需要进一步研究。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

求助全文

约1分钟内获得全文求助全文

来源期刊

Life sciences 医学-药学

CiteScore

12.20

自引率

1.60%

发文量

841

审稿时长

6 months

期刊介绍： Life Sciences is an international journal publishing articles that emphasize the molecular, cellular, and functional basis of therapy. The journal emphasizes the understanding of mechanism that is relevant to all aspects of human disease and translation to patients. All articles are rigorously reviewed. The Journal favors publication of full-length papers where modern scientific technologies are used to explain molecular, cellular and physiological mechanisms. Articles that merely report observations are rarely accepted. Recommendations from the Declaration of Helsinki or NIH guidelines for care and use of laboratory animals must be adhered to. Articles should be written at a level accessible to readers who are non-specialists in the topic of the article themselves, but who are interested in the research. The Journal welcomes reviews on topics of wide interest to investigators in the life sciences. We particularly encourage submission of brief, focused reviews containing high-quality artwork and require the use of mechanistic summary diagrams.