Antioxidant effect of esculin and sulforaphane in zebrafish larvae: A comparative study

IF 3.9 3区环境科学与生态学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Comparative Biochemistry and Physiology C-toxicology & Pharmacology Pub Date : 2025-07-03 DOI:10.1016/j.cbpc.2025.110264

Srija Babu, Vignesh Sundarraja, Suryaa Manoharan, Ekambaram Perumal

{"title":"Antioxidant effect of esculin and sulforaphane in zebrafish larvae: A comparative study","authors":"Srija Babu, Vignesh Sundarraja, Suryaa Manoharan, Ekambaram Perumal","doi":"10.1016/j.cbpc.2025.110264","DOIUrl":null,"url":null,"abstract":"<div><div>Antioxidants are crucial for maintaining cellular redox homeostasis by neutralizing free radicals, including reactive oxygen species (ROS). Nuclear factor erythroid-2-related factor 2 (Nrf2) is a transcription factor that activates various cytoprotective genes to counteract oxidative damage. The present study aimed to compare the antioxidant potential of natural phytochemicals, esculin (ESC), and sulforaphane (SFN) in zebrafish larvae, focusing on Nrf2 activation. Zebrafish larvae were treated with ESC (100 μM) and SFN (40 μM) along with control for 6 h. The levels of ROS, lipid peroxidation (LPO), nitric oxide (NO), glutathione S-transferase (GST), glutathione peroxidase (GPx), catalase (CAT), and reduced glutathione (GSH) were measured. Further, RT-PCR and Western blotting methods were employed to determine the antioxidant genes and protein levels, respectively. In addition, Nrf2 localization within the cell was assessed using nuclear/cytosolic fractionation assay and wholemount immunohistochemistry method. Our results demonstrate that ESC treatment significantly increased antioxidant enzyme activities (GST, GPx, CAT), similar to the effects observed with SFN. Further, ESC enhanced the mRNA levels of antioxidant genes expression, such as <em>nrf2, gstp1, hmox1a, prdx1, nqo1, gss, gsr, sqstm1, hsp90aa1.2</em> than SFN. Antioxidant proteins Nrf2, Gstp1, Cat, and Sod2 were upregulated in the ESC-treated groups. Subsequently, nuclear localization of Nrf2 was detected in both SFN- and ESC-treated groups, confirming Nrf2 translocation into the nucleus. Our findings suggest that ESC possesses potent antioxidant activity as SFN in 72-hpf zebrafish larvae through the Keap1-Nrf2 signaling pathway.</div></div>","PeriodicalId":10602,"journal":{"name":"Comparative Biochemistry and Physiology C-toxicology & Pharmacology","volume":"297 ","pages":"Article 110264"},"PeriodicalIF":3.9000,"publicationDate":"2025-07-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Comparative Biochemistry and Physiology C-toxicology & Pharmacology","FirstCategoryId":"93","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S1532045625001450","RegionNum":3,"RegionCategory":"环境科学与生态学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"BIOCHEMISTRY & MOLECULAR BIOLOGY","Score":null,"Total":0}

引用次数: 0

Abstract

Antioxidants are crucial for maintaining cellular redox homeostasis by neutralizing free radicals, including reactive oxygen species (ROS). Nuclear factor erythroid-2-related factor 2 (Nrf2) is a transcription factor that activates various cytoprotective genes to counteract oxidative damage. The present study aimed to compare the antioxidant potential of natural phytochemicals, esculin (ESC), and sulforaphane (SFN) in zebrafish larvae, focusing on Nrf2 activation. Zebrafish larvae were treated with ESC (100 μM) and SFN (40 μM) along with control for 6 h. The levels of ROS, lipid peroxidation (LPO), nitric oxide (NO), glutathione S-transferase (GST), glutathione peroxidase (GPx), catalase (CAT), and reduced glutathione (GSH) were measured. Further, RT-PCR and Western blotting methods were employed to determine the antioxidant genes and protein levels, respectively. In addition, Nrf2 localization within the cell was assessed using nuclear/cytosolic fractionation assay and wholemount immunohistochemistry method. Our results demonstrate that ESC treatment significantly increased antioxidant enzyme activities (GST, GPx, CAT), similar to the effects observed with SFN. Further, ESC enhanced the mRNA levels of antioxidant genes expression, such as nrf2, gstp1, hmox1a, prdx1, nqo1, gss, gsr, sqstm1, hsp90aa1.2 than SFN. Antioxidant proteins Nrf2, Gstp1, Cat, and Sod2 were upregulated in the ESC-treated groups. Subsequently, nuclear localization of Nrf2 was detected in both SFN- and ESC-treated groups, confirming Nrf2 translocation into the nucleus. Our findings suggest that ESC possesses potent antioxidant activity as SFN in 72-hpf zebrafish larvae through the Keap1-Nrf2 signaling pathway.

Abstract Image

查看原文本刊更多论文

esculin和萝卜硫素对斑马鱼幼鱼抗氧化作用的比较研究。

抗氧化剂是通过中和自由基（包括活性氧）来维持细胞氧化还原稳态的关键。核因子红细胞2相关因子2- Nrf2是一种激活多种细胞保护基因以对抗氧化损伤的转录因子。本研究旨在比较天然植物化学物质esculin （ESC）和sulforaphane （SFN）在斑马鱼幼虫中的抗氧化能力，重点研究Nrf2的激活。分别用ESC（100 μM）和SFN（40 μM）处理斑马鱼幼鱼6 h。测定小鼠体内ROS、脂质过氧化（LPO）、一氧化氮（NO）、谷胱甘肽s -转移酶（GST）、谷胱甘肽过氧化物酶（GPx）、过氧化氢酶（CAT）、还原性谷胱甘肽（GSH）水平。采用RT-PCR和Western blotting方法分别检测抗氧化基因和蛋白水平。此外，采用核/胞浆分离法和全量免疫组织化学方法评估Nrf2在细胞内的定位。结果表明，ESC处理显著提高了抗氧化酶活性（GST、GPx、CAT），与SFN处理的效果相似。此外，与SFN相比，ESC提高了nrf2、gstp1、hmox1a、prdx1、nqo1、gss、gsr、sqstm1、hsp90aa1.2等抗氧化基因的mRNA表达水平。抗氧化蛋白Nrf2、Gstp1、Cat和Sod2在esc处理组上调。随后，在SFN-和esc处理组中均检测到Nrf2的核定位，证实Nrf2易位到细胞核中。我们的研究结果表明，ESC作为SFN通过Keap1-Nrf2信号通路在72-hpf斑马鱼幼虫中具有强大的抗氧化活性。

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

Comparative Biochemistry and Physiology C-toxicology & Pharmacology 医学-动物学

CiteScore

7.50

自引率

5.10%

发文量

206

审稿时长

30 days

期刊介绍： Part C: Toxicology and Pharmacology. This journal is concerned with chemical and drug action at different levels of organization, biotransformation of xenobiotics, mechanisms of toxicity, including reactive oxygen species and carcinogenesis, endocrine disruptors, natural products chemistry, and signal transduction with a molecular approach to these fields.