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两种毒液变体的酶活性、凝血活性和神经肌肉活性及抗蛇毒血清疗效的比较分析

Comparative Analysis of the Enzymatic, Coagulant, and Neuromuscular Activities of Two Variants of Venom and Antivenom Efficacy.

作者信息

Demico Poliana J, Oliveira Isabele N, Proença-Hirata Vitória S, Dias Samuel R, Ghirotti Hugo A, Silva Elisangela O, Giometti Inês C, Pacagnelli Francis L, Torres-Bonilla Kristian A, Hyslop Stephen, Galizio Nathália C, de Morais-Zani Karen, Pucca Manuela B, Rocha Anderson M, Maciel Jéssica B, Sartim Marco A, Monteiro Wuelton M, Floriano Rafael S

机构信息

Laboratory of Toxinology and Cardiovascular Research, University of Western São Paulo (UNOESTE), Presidente Prudente 19050-680, SP, Brazil.

Department of Pharmacology, Faculty of Medical Sciences, State University of Campinas (UNICAMP), Campinas 13083-888, SP, Brazil.

出版信息

Pharmaceuticals (Basel). 2025 Jan 6;18(1):54. doi: 10.3390/ph18010054.

Abstract

We compared the enzymatic, coagulant, and neuromuscular activities of two variants (yellow-CDRy and white-CDRw) of venom with a sample of (CDT) venom and examined their neutralization by antivenom against CDT venom. The venoms were screened for enzymatic and coagulant activities using standard assays, and electrophoretic profiles were compared by SDS-PAGE. Neutralization was assessed by preincubating venoms with crotalic antivenom and assaying the residual activity. SDS-PAGE showed that the venoms had similar electrophoretic profiles, with the main bands being phospholipase A (PLA), serine proteinases, L-amino acid oxidase (LAAO), and phosphodiesterase. CDRy venom had the highest proteolytic and LAAO activities, CDRw venom had greater PLA and esterolytic activities at the highest quantity tested, and CDT had greater PLA activity than CDRy. CDRw and CDT venoms had similar proteolytic and LAAO activities, and CDRy and CDT venoms had comparable esterolytic activity. None of the venoms altered the prothrombin time (PT), but all of them decreased the activated partial thromboplastin time (aPPT); this activity was neutralized by antivenom. The minimum coagulant dose potency was CDRw >> CDRy > CDT. All venoms had thrombin-like activity that was attenuated by antivenom. CDRy and CDRw venoms showed α-fibrinogenolytic activity. All venoms partially cleaved the β-chain. CDRy and CDT venoms caused neuromuscular facilitation (enhanced muscle contractions) followed by complete blockade, whereas CDRw venom caused only blockade. Antivenom neutralized the neuromuscular activity to varying degrees. These findings indicate that while CDR and CDT venoms share similarities, they also differ in some enzymatic and biological activities and in neutralization by antivenom. Some of these differences could influence the clinical manifestations of envenomation by and their neutralization by the currently used therapeutic antivenom.

摘要

我们将两种毒液变体(黄色 - CDRy和白色 - CDRw)的酶活性、凝血活性和神经肌肉活性与一种(CDT)毒液样本进行了比较,并检测了抗CDT毒液血清对它们的中和作用。使用标准测定法对毒液的酶活性和凝血活性进行筛选,并通过SDS - PAGE比较电泳图谱。通过将毒液与抗蝮蛇毒血清预孵育并测定残余活性来评估中和作用。SDS - PAGE显示,毒液具有相似的电泳图谱,主要条带为磷脂酶A(PLA)、丝氨酸蛋白酶、L - 氨基酸氧化酶(LAAO)和磷酸二酯酶。CDRy毒液具有最高的蛋白水解和LAAO活性,在测试的最高剂量下,CDRw毒液具有更高的PLA和酯水解活性,并且CDT的PLA活性高于CDRy。CDRw和CDT毒液具有相似的蛋白水解和LAAO活性,CDRy和CDT毒液具有相当的酯水解活性。所有毒液均未改变凝血酶原时间(PT),但它们均缩短了活化部分凝血活酶时间(aPPT);这种活性被抗蛇毒血清中和。最小凝血剂量效力为CDRw >> CDRy > CDT。所有毒液都具有类似凝血酶的活性,该活性被抗蛇毒血清减弱。CDRy和CDRw毒液表现出α - 纤维蛋白溶解活性。所有毒液都部分切割了β链。CDRy和CDT毒液引起神经肌肉易化(增强肌肉收缩),随后完全阻断,而CDRw毒液仅引起阻断。抗蛇毒血清不同程度地中和了神经肌肉活性。这些发现表明,虽然CDR和CDT毒液有相似之处,但它们在一些酶活性和生物学活性以及抗蛇毒血清的中和作用方面也存在差异。其中一些差异可能会影响由它们引起的中毒的临床表现以及目前使用的治疗性抗蛇毒血清对其的中和作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6bba/11768973/b9ac9e2d774f/pharmaceuticals-18-00054-g001.jpg

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