Ferreira Felipe Roberto Borba, da Silva Pollyanna Michelle, Soares Tatiana, Gonçalves Machado Larissa, de Araújo Larissa Cardoso Corrêa, da Silva Teresinha Gonçalves, de Mello Gabriela Souto Vieira, Galdino da Rocha Pitta Maira, de Melo Rego Moacyr Jesus Barreto, Pontual Emmanuel Viana, Zingali Russolina Benedeta, Napoleão Thiago Henrique, Paiva Patrícia Maria Guedes
Departamento de Bioquímica, Centro de Biociências, Universidade Federal de Pernambuco, Recife, PE, Brazil.
Centro de Tecnologias Estratégicas do Nordeste, Recife, PE, Brazil.
Toxicon. 2016 Nov;122:119-126. doi: 10.1016/j.toxicon.2016.09.019. Epub 2016 Sep 29.
This study characterized the protein/peptide profile of venom isolated from the spider Lasiodora sp. (Mygalomorphae, Theraphosidae) found in northeastern Brazil and determined its antimicrobial activity, toxicity against human cells, and hemolytic activity. Protein concentration of the Lasiodora sp. venom was 4.53 ± 0.38 mg/mL. SDS-PAGE showed proteins with molecular masses up to 75 kDa, some of which contained disulfide bridges. RP-HPLC analysis separate at least 12 peaks that were identified by mass spectrometry as peptides U-theraphotoxin-Lp1a (lasiotoxin-1), U-theraphotoxin-Lp1c (lasiotoxin-3), U-theraphotoxin-Lsp1a (LTx5), and ω-theraphotoxin-Asp3a as well as the proteins phospholipase A (PLA) and hyaluronidase. The crude venom exhibited bactericidal effect against Aeromonas sp., Bacillus subtilis, and Micrococcus luteus and fungicidal effect against Candida parapsilosis and Candida albicans. In addition, the venom exerted bacteriostatic effect against Klebsiella pneumoniae, Pseudomonas aeruginosa, and Staphylococcus aureus and fungistatic effect against Candida tropicalis and Candida krusei. The minimum inhibitory (MIC), minimum bactericidal (MBC), and minimum fungicidal (MFC) concentrations ranged from 3.9 to 500 μg/mL. The Lasiodora sp. venom decreased the viability of human peripheral blood mononuclear cells (PBMCs) by 50%-90% at concentrations of 0.1, 1, 10, and 100 μg/mL, promoting apoptosis of these cells. On the other hand, the venom showed weak hemolytic activity against Mus musculus erythrocytes (EC: 757 μg/mL). In conclusion, the Lasiodora sp. spider venom is a rich source of antimicrobial agents. Future studies will focus on identifying antimicrobial agents present in this venom and evaluating whether these agents contribute to its cytotoxic effects against PBMCs.
本研究对从巴西东北部发现的蜘蛛Lasiodora sp.（原蛛亚目，捕鸟蛛科）分离出的毒液的蛋白质/肽谱进行了表征，并测定了其抗菌活性、对人细胞的毒性和溶血活性。Lasiodora sp.毒液的蛋白质浓度为4.53±0.38mg/mL。SDS-PAGE显示分子量高达75kDa的蛋白质，其中一些含有二硫键。RP-HPLC分析分离出至少12个峰，通过质谱鉴定为肽U-theraphotoxin-Lp1a（lasiotoxin-1）、U-theraphotoxin-Lp1c（lasiotoxin-3）、U-theraphotoxin-Lsp1a（LTx5）和ω-theraphotoxin-Asp3a以及蛋白质磷脂酶A（PLA）和透明质酸酶。粗毒液对气单胞菌属、枯草芽孢杆菌和藤黄微球菌具有杀菌作用，对近平滑念珠菌和白色念珠菌具有杀真菌作用。此外，毒液对肺炎克雷伯菌、铜绿假单胞菌和金黄色葡萄球菌具有抑菌作用，对热带念珠菌和克柔念珠菌具有抑真菌作用。最低抑菌（MIC）、最低杀菌（MBC）和最低杀真菌（MFC）浓度范围为3.9至500μg/mL。Lasiodora sp.毒液在0.1、1、10和100μg/mL浓度下可使人类外周血单核细胞（PBMC）的活力降低50%-90%，促进这些细胞的凋亡。另一方面，该毒液对小家鼠红细胞的溶血活性较弱（半数有效浓度：757μg/mL）。总之，Lasiodora sp.蜘蛛毒液是抗菌剂的丰富来源。未来的研究将集中于鉴定该毒液中存在的抗菌剂，并评估这些药剂是否对其对PBMC的细胞毒性作用有贡献。
