S-亚硝基谷胱甘肽提高曲霉孢子抗氧化能力

S-Nitrosoglutathione Enhances the Antioxidant Capacity ofAspergillus nidulans Conidia

摘要

NO 和 H₂O₂ 是免疫系统产生的两类活性抗菌分子，各自具有不同的细胞毒性。虽然通常认为它们的共同存在会增强抗菌效应，但已有证据表明，某些动物细胞、植物细胞以及细菌能够利用 NO 快速缓解氧化损伤，从而抵消这种效应。然而，在丝状真菌中，NO 在 H₂O₂ 耐受性调控中的作用尚未得到广泛研究。本研究探讨了 NO 供体 S-亚硝基谷胱甘肽（GSNO）对构巢曲霉（Aspergillus nidulans）分生孢子 H₂O₂ 耐受性的影响。结果表明，GSNO 预处理显著提高了休眠分生孢子在 2 mmol/L H₂O₂ 压力下的存活率，而对膨润孢子和菌丝无显著作用；通过关键氧化应激抗性基因的敲除与表型分析，鉴定出过氧还蛋白（PrxA）是该增强效应的核心介导因子；GSNO 预处理促进了 PrxA 二聚体中二硫键的还原，导致 H₂O₂ 压力下胞内 PrxA 二聚体水平下降，这一修饰降低了 PrxA 催化分解 H₂O₂ 的能力，从而节约胞内还原力，并将细胞资源重新分配至蛋白质和 DNA 的氧化损伤修复。此研究结果为真菌分生孢子抵御氧化压力的保护机制提供了新的见解，并揭示了 NO 在真菌应激中新的作用。

Abstract

Nitric oxide (NO) and hydrogen peroxide (H₂O₂) are two reactive antimicrobial molecules produced by the immune system, each possessing distinct cytotoxic properties. While their combined presence is generally believed to enhance antimicrobial efficacy, evidence suggests that certain animal and plant cells, as well as bacteria, can counteract this effect by utilizing NO to rapidly mitigate oxidative damage. However, the role of NO in modulating H₂O₂ resistance in filamentous fungi has not been extensively studied. In this study, we investigated the effect of the NO donor S-nitrosoglutathione (GSNO) on H₂O₂ resistance ofAspergillus nidulans conidia. We found that GSNO pre-treatment significantly enhanced the survival rate of dormant conidia exposed to 2 mmol/L H₂O₂, while having no effect on swollen conidia or hyphae. Through knockout and phenotypic analysis of key oxidative stress resistance genes, we identified peroxiredoxin (PrxA) as a critical mediator of this enhanced resistance. Further biochemical analyses revealed that GSNO pre-treatment promoted the reduction of disulfide bonds in PrxA dimers, leading to decreased intracellular PrxA dimer levels under H₂O₂ stress. This modification reduced PrxA-mediated H₂O₂ decomposition, thereby conserving intracellular reducing power and redirecting cellular resources toward the repair of oxidative damage to proteins and DNA. Our findings provide novel insights into the protective mechanisms employed by fungal conidia to withstand oxidative stress and highlight a previously unrecognized role of NO in fungal stress adaptation.

关键词

NO / H₂O₂ / 构巢曲霉 / 过氧还蛋白 / 氧化压力 / S-亚硝基谷胱甘肽

Key words

NO / H₂O₂ / Aspergillus nidulans / peroxiredoxin (PrxA) / oxidative stress / S-nitrosoglutathione (GSNO)

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基金资助

国家自然科学基金(22077032; 21672065)

天然药物及仿生药物国家重点实验室(K202415)