中华眼镜蛇毒致广西巴马小型猪坏死组织渗出液中的蛇毒成分分析

Analysis of the venom components of necrotic tissue exudates from Naja atra venom-infected Guangxi Bama miniature pigs

  • 摘要:
    目的  探究中华眼镜蛇毒致广西巴马小型猪坏死组织渗出液中的蛇毒成分及其蛋白表达的动态变化。
    方法  给予广西巴马小型猪注射中华眼镜蛇毒,构建蛇伤中毒的局部组织坏死模型,在注射毒素后的6 h、12 h、24 h、36 h和48 h取小猪坏死组织的渗出液,采用无标记的蛋白质组学技术鉴定分析渗出液中蛇毒蛋白质组成及动态变化,通过对所鉴定出的蛋白进行KEGG及GO通路富集分析,以深入了解蛇毒蛋白相关的生物学功能。
    结果  通过蛇伤中毒后广西巴马小型猪的生物学行为、局部肌肉组织的病理结果、注射部位的伤口变化来评判动物模型,广西巴马小型猪中毒后出现呼吸急促、肌肉组织坏死、局部伤口溃烂等症状,均符合临床特征的实际情况,表明成功构建了中华眼镜蛇毒导致局部组织坏死的广西巴马小型猪模型。在广西巴马小型猪的局部组织坏死渗出液中鉴定到40种蛇毒蛋白质,涵盖三指毒素、磷脂酶A2、富含半胱氨酸分泌蛋白、蛇毒金属蛋白酶、核苷酸焦磷酸/磷酸二酯酶等蛇毒蛋白家族,并呈现出动态性数量变化。
    结论  在中华眼镜蛇毒致广西巴马小型猪过程中,不同时间段的组织渗出液中均鉴定出多种三指毒素、磷脂酶A2等蛇毒蛋白,其可能在局部组织坏死中发挥着重要作用。

     

    Abstract:
    Objective  To investigate the components of snake venom and the dynamic changes in protein expression in the exudates of necrotic tissues affected by the Naja atra venom in Guangxi Bama miniature pigs.
    Methods  Guangxi Bama miniature pigs were injected with Naja atra venom to establish a local tissue necrosis model due to snakebite poisoning. Exudates from the necrotic tissues of the pigs were collected at 6 h, 12 h, 24 h, 36 h, and 48 h post-injection. Label-free proteomics techniques were employed to identify and analyze the composition and dynamic changes of snake venom proteins in the exudates. KEGG and GO pathway enrichment analyses were performed on the identified proteins to gain deeper insights into the biological functions related to snake venom proteins.
    Results  The biological behavior of the Guangxi Bama miniature pigs post-snakebite, pathological results of local muscle tissues, and changes at the injection site were used to evaluate the animal model. The pigs exhibited symptoms such as rapid respiration, muscle tissue necrosis, and local wound ulceration, consistent with clinical characteristics, indicating the successful establishment of a Guangxi Bama miniature pig model of local tissue necrosis induced by Naja atra venom. A total of 40 snake venom proteins were identified in the exudates of the local tissue necrosis in the Guangxi Bama miniature pigs, including three-finger toxins, phospholpase A2, cysteine- rich secretory proteins, snake venom metalloproteinases, and nucleotidyl pyrophosphatase/phosphodiesterases, showing dynamic quantitative changes.
    Conclusion  Various snake venom proteins, including three-finger toxins and phospholipase A2, are identified in the exudates of the tissues at different time points during the poisoning process of Guangxi Bama miniature pigs by Naja atra venom, suggesting their important roles in local tissue necrosis.

     

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