Ivanović Saša R, Rešetar Maslov Dina, Rubić Ivana, Mrljak Vladimir, Živković Irena, Borozan Nevena, Grujić-Milanović Jelica, Borozan Sunčica
Department of Pharmacology and Toxicology, Faculty of Veterinary Medicine, University of Belgrade, Bulevar Oslobođenja 18, 11000 Belgrade, Serbia.
Laboratory of Proteomics, Clinic for Internal Diseases, Faculty of Veterinary Medicine, University of Zagreb, Heinzelova 55, 10000 Zagreb, Croatia.
Vet Sci. 2024 Nov 28;11(12):605. doi: 10.3390/vetsci11120605.
Deep proteomic analyses identified, in total, 159 master proteins (with 1% FDR and 2 unique peptides) from 26 protein families in the venom of Data are available via ProteomeXchange with the identifier PXD056495. The relative abundance of PLA2s is 11.60% of the crude venom, of which 4.35% are neurotoxic Ammodytoxins (Atxs). The neurotoxicity of the venom of and the neutralizing effect of the antivenom were tested on the neuromuscular preparation of the diaphragm (NPD) of rats. The activity of PLA2 in the venom of and its neutralization by the antivenom were determined under in vitro conditions. The venom leads to a progressive decrease in NPD contractions. We administered pre-incubated venom/antivenom mixtures at various ratios of 1:2, 1:10 and 1:20 (/) and observed the effects of these mixtures on NPD contractions. The results show that the mean effective time (ET) for NPD contractions with the 1:20 mixture is highly significantly different ( < 0.001) from the ET for the venom and the ET for the 1:2 and 1:10 mixture ratios. We also found a highly significant ( < 0.001) reduction in Na/K-ATPase activity in the NPD under the influence of the venom. The reduction in the activity of this enzyme was reversible by the antivenom. Under in vitro conditions, we have achieved the complete neutralization of PLA2 by the antivenom. In conclusion, the antivenom abolished the venom-induced progressive decrease in NPD contractions in a concentration-dependent manner. Antivenom with approximately the same mass proportion almost completely restores Na/K-ATPase activity in the NPD and completely neutralizes the PLA2 activity of the venom in vitro.
深度蛋白质组学分析总共从[具体物种]毒液中的26个蛋白质家族中鉴定出159种主蛋白(错误发现率为1%且有2条独特肽段)。数据可通过ProteomeXchange获取,标识符为PXD056495。磷脂酶A2(PLA2s)的相对丰度占粗毒液的11.60%,其中4.35%是神经毒性的锯鳞蝰毒素(Atxs)。[具体物种]毒液的神经毒性及抗蛇毒血清的中和作用在大鼠膈神经肌肉标本(NPD)上进行了测试。在体外条件下测定了[具体物种]毒液中PLA2的活性及其被抗蛇毒血清的中和情况。[具体物种]毒液导致NPD收缩逐渐减少。我们以1:2、1:10和1:20的不同比例(毒液/抗蛇毒血清)给予预孵育的毒液/抗蛇毒血清混合物,并观察这些混合物对NPD收缩的影响。结果表明,1:20混合物使NPD收缩的平均有效时间(ET)与毒液的ET以及1:2和1:10混合比例的ET相比有极显著差异(P<0.001)。我们还发现,在毒液影响下,NPD中钠钾ATP酶活性有极显著降低(P<0.001)。该酶活性的降低可被抗蛇毒血清逆转。在体外条件下,抗蛇毒血清实现了对PLA2的完全中和。总之,抗蛇毒血清以浓度依赖的方式消除了毒液诱导的NPD收缩逐渐减少的现象。质量比例大致相同的抗蛇毒血清几乎完全恢复了NPD中的钠钾ATP酶活性,并在体外完全中和了毒液的PLA2活性。