Micronutrient antioxidant supplementation alleviates valproic acid-induced oxidative stress and male infertility via the NRF2/HO-1 pathway

IF 10.7 1区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Redox Biology Pub Date : 2025-05-29 DOI:10.1016/j.redox.2025.103685

Muhammad Arif Asghar , Bing Wan , Lu Li , Jie Zhang , Shixin Tang , Hang Han , Yuanyuan Yang , Long Chu , Qian Zhang , Xiao Zhang , Qinjian Zhao

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引用次数: 0

Abstract

Background

Valproic Acid (VPA), a widely used anticonvulsant, is known to induce oxidative stress, contributing to male infertility. This study explores the potential of micronutrient antioxidants to improve fertility in VPA-treated individuals.

Methods

Six-week-old male mice were treated with VPA and supplemented with antioxidants, including l-Arginine (120 mg/kg), N-Acetylcysteine (NAC) (2 mg/kg), Taurine (200 mg/kg), L-Tryptophan (0.5 mg/kg), Zinc chloride (ZnCl2) (1.5 mg/kg), and Selenium (0.5 mg/kg). The dosing regimen lasted for 34 days. Sperm quality, oxidative stress, and inflammatory biomarkers were assessed through gene expression analysis, western blotting, histological assessments, TUNEL assays, and immunohistochemistry. Additionally, GC-2spd(ts) and HepG2 cell lines were used to examine the testicular and systemic effects of VPA and antioxidants. Network pharmacology was applied to identify key molecular targets and pathways.

Results

Antioxidant supplementation significantly improved sperm count, with l-Arginine showing an approximately 296.1 % increase, NAC a 270.7 % increase, and Taurine a 255.9 % increase compared to the VPA-only group. Furthermore, antioxidants enhanced semen volume, testosterone levels, sperm motility, morphology, and viability. Gene expression analysis revealed significant upregulation of key oxidative stress-related proteins such as SOD1, HO-1, NRF2, and NQO1. Western blot and histological analyses showed a reversal of oxidative stress and preservation of seminiferous tubule integrity. TUNEL assays demonstrated a reduction in apoptotic damage, and IHC confirmed an increase in HO-1 and SOD1. In vitro studies with GC-2spd(ts) and HepG2 cells confirmed that antioxidants alleviated VPA-induced oxidative stress. Network pharmacology identified key molecular targets, such as GPX4, SOD1, HO-1, and NRF2, which are involved in oxidative stress, apoptosis, and inflammation pathways, that were modulated by antioxidants.

Conclusion

Micronutrient antioxidants effectively reduce VPA-induced oxidative stress and improve male fertility. These results suggest that antioxidant supplementation could be a promising strategy to mitigate oxidative damage and enhance fertility in individuals undergoing VPA therapy.

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补充微量营养素抗氧化剂可通过NRF2/HO-1途径缓解丙戊酸诱导的氧化应激和男性不育

丙戊酸（VPA）是一种广泛使用的抗惊厥药，已知会引起氧化应激，导致男性不育。本研究探讨了微量营养素抗氧化剂提高vpa治疗个体生育能力的潜力。方法6周龄雄性小鼠经VPA处理后，添加抗氧化剂：l-精氨酸（120 mg/kg）、n -乙酰半胱氨酸（NAC）（2 mg/kg）、牛磺酸（200 mg/kg）、l-色氨酸（0.5 mg/kg）、氯化锌（ZnCl2）（1.5 mg/kg）、硒（0.5 mg/kg）。给药期为34 d。通过基因表达分析、western blotting、组织学评估、TUNEL检测和免疫组织化学来评估精子质量、氧化应激和炎症生物标志物。此外，GC-2spd（ts）和HepG2细胞系被用来检测VPA和抗氧化剂对睾丸和全身的影响。网络药理学应用于鉴定关键分子靶点和通路。结果：与纯vpa组相比，补充抗氧化剂显著提高了精子数量，l-精氨酸增加了约296.1%，NAC增加了270.7%，牛磺酸增加了255.9%。此外，抗氧化剂还能增加精液量、睾酮水平、精子活力、形态和活力。基因表达分析显示SOD1、HO-1、NRF2和NQO1等关键氧化应激相关蛋白显著上调。Western blot和组织学分析显示氧化应激的逆转和精管完整性的保存。TUNEL实验显示凋亡损伤减少，IHC证实HO-1和SOD1增加。GC-2spd（ts）和HepG2细胞的体外研究证实，抗氧化剂可减轻vpa诱导的氧化应激。网络药理学鉴定出GPX4、SOD1、HO-1和NRF2等关键分子靶点，它们参与氧化应激、细胞凋亡和炎症通路，并受抗氧化剂调节。结论微量营养素抗氧化剂能有效降低vpa诱导的氧化应激，提高男性生育能力。这些结果表明，在接受VPA治疗的个体中，补充抗氧化剂可能是一种有希望的减轻氧化损伤和提高生育能力的策略。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Redox Biology BIOCHEMISTRY & MOLECULAR BIOLOGY-

CiteScore

19.90

自引率

3.50%

发文量

318

审稿时长

25 days

期刊介绍： Redox Biology is the official journal of the Society for Redox Biology and Medicine and the Society for Free Radical Research-Europe. It is also affiliated with the International Society for Free Radical Research (SFRRI). This journal serves as a platform for publishing pioneering research, innovative methods, and comprehensive review articles in the field of redox biology, encompassing both health and disease. Redox Biology welcomes various forms of contributions, including research articles (short or full communications), methods, mini-reviews, and commentaries. Through its diverse range of published content, Redox Biology aims to foster advancements and insights in the understanding of redox biology and its implications.