神經壞死病毒（Nervous necrosis virus, NNV）為石斑魚養殖產業中主要的病原之一，常引發大腦組織損傷與高死亡率。然而，NNV 感染所誘發的免疫病理機制仍未被完全釐清。近年研究指出，程序性壞死（necroptosis）為一種免疫相關的細胞死亡路徑，可能參與病毒感染引起的免疫失衡與組織傷害。因此，本研究旨在探討 NNV 感染是否透過活化 necroptosis 路徑，促發石斑魚腦部免疫病理反應，並進一步探討抗氧化劑對相關路徑的調控作用。
本研究以 GF-1 細胞與石斑魚苗進行 NNV 感染模型，結果顯示在感染後 RIPK1、RIPK3 與 MLKL 基因與蛋白表現量均明顯上升，轉錄體分析亦顯示感染組在 necroptosis 及多項免疫因子（如 IL-1β、IL-10、IL-21）表現上調，顯示 NNV 可能促發細胞死亡與免疫風暴。此外，抗氧化劑處理可降低 RIPK1、RIPK3 表現，並抑制 oxidative stress 相關基因（如 Prdx、NOX、SOD）與部分發炎反應，顯示 ROS 在此病理機制中具調節角色。
綜合上述結果，本研究首次系統性證實 NNV 感染可透過誘發 necroptosis 路徑活化，促進石斑魚腦部免疫反應失衡與發炎傷害，並指出氧化壓力在此過程中可能具有調控角色。此結果有助於釐清 NNV 感染之致病機轉，並為未來魚類病毒疾病的免疫調控與治療策略提供重要參考依據。

Nervous necrosis virus (NNV) is a highly neurotropic pathogen responsible for severe brain lesions and high mortality in groupers. While its pathological effects are well-documented, the underlying mechanisms of virus-induced immune and inflammatory responses remain unclear. This study investigates whether NNV infection activates necroptosis—a form of programmed necrotic cell death—and how this process contributes to immune dysregulation and inflammatory pathology.
Through qPCR and Western blot analyses in GF-1 cells and infected grouper brain tissues, we observed significant upregulation of key necroptosis regulators: RIPK1, RIPK3, and MLKL. Transcriptomic profiling further revealed the concurrent induction of necroptosis-related genes and inflammatory cytokines, including IL-1β, IL-10, and IL-21, suggesting a link between virus-induced cell death and cytokine imbalance. These findings support the hypothesis that NNV-induced necroptosis may act as a driver of immune overactivation and contribute to cytokine storm-like conditions in the infected host.
Additionally, we evaluated the potential modulatory effect of antioxidant treatment, which partially suppressed necroptosis-related gene expression and inflammatory markers, implying a possible regulatory role of oxidative stress in this process.
Together, these results provide the first evidence that NNV can induce necroptosis-driven immunopathology in grouper brain tissue, offering novel insights into the cellular mechanisms underlying fish viral encephalopathy and potential directions for immunoregulatory intervention.

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