College of Animal Science and Technology, Northwest A&F University, Xinong Road 22nd, Xianyang 712100, China.
Int J Mol Sci. 2022 Sep 9;23(18):10448. doi: 10.3390/ijms231810448.
The outbreak of white spot syndrome (WSS) is a looming challenge, due to dramatic losses to the crustacean aquaculture industry. However, at present, there are no prophylactic or therapeutic means to control this infectious viral disease. Here, we screened fifteen medicinal plants for their inhibitory activity on the white spot syndrome virus (WSSV), using red swamp crayfish (Procambarus clarkii) as a model species. The results showed that the crude extracts of Pinellia ternata (Thunb.) Breit. had the highest inhibitory effect (91.59%, 100 mg/kg) on WSSV proliferation, and its main component, beta-sitosterol, showed a much higher activity (95.79%, 50 mg/kg). Further, beta-sitosterol potently reduced (p < 0.01) viral loads and viral gene transcription levels in a concentration-dependent fashion, and significantly promoted the survival rate of WSSV-challenged crayfish (57.14%, 50 mg/kg). The co-incubation assay indicated that beta-sitosterol did not influence the infectivity of WSSV particles. Both pre- and post-treatment of beta-sitosterol exerted a significant inhibitory effect (p < 0.01) on the viral load in vivo. Mechanistically, beta-sitosterol not only interfered with the expression of viral genes (immediate early gene 1, ie1; DNA polymerase, DNApol) that are important in initiating WSSV transcription, but it also attenuated the hijacking of innate immune signaling pathways (Toll, IMD, and JAK/STAT pathways) by viral genes to block WSSV replication. Moreover, the expression of several antiviral immune, antioxidant, pro-inflammatory, and apoptosis-related genes changed significantly in beta-sitosterol-treated crayfish. Beta-sitosterol is a potent WSSV inhibitor and has the potential to be developed as an effective anti-WSSV agent against a WSS outbreak in crustacean aquaculture.
白斑综合征（WSS）的爆发是甲壳类水产养殖业面临的一个严峻挑战，因为它会导致严重的损失。然而，目前尚无预防或治疗这种传染性病毒疾病的方法。在这里，我们以红螯螯虾（Procambarus clarkii）为模型物种，筛选了 15 种药用植物对白斑综合征病毒（WSSV）的抑制活性。结果表明，半夏（Pinellia ternata (Thunb.) Breit.）的粗提取物对 WSSV 增殖的抑制作用最强（91.59%，100mg/kg），其主要成分β-谷甾醇的活性更高（95.79%，50mg/kg）。此外，β-谷甾醇能显著降低（p<0.01）病毒载量和病毒基因转录水平，并显著提高 WSSV 感染虾的存活率（57.14%，50mg/kg）。共孵育试验表明，β-谷甾醇不影响 WSSV 颗粒的感染力。β-谷甾醇的预治疗和后治疗均对体内病毒载量有显著抑制作用（p<0.01）。在机制上，β-谷甾醇不仅干扰了 WSSV 转录起始过程中重要的早期基因 1（ie1）和 DNA 聚合酶（DNApol）等病毒基因的表达，而且还减弱了病毒基因对先天免疫信号通路（Toll、IMD 和 JAK/STAT 通路）的劫持，从而阻断了 WSSV 的复制。此外，β-谷甾醇处理的虾体内几种抗病毒免疫、抗氧化、促炎和凋亡相关基因的表达发生了显著变化。β-谷甾醇是一种有效的 WSSV 抑制剂，有潜力开发为防治甲壳类水产养殖中 WSS 爆发的有效抗 WSSV 药物。
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