Tavares Maria Naiara Macedo, de Melo Cordeiro Eulalio Micaela, Santana Hallison Mota, Boeno Charles Nunes, Dos Reis Valdison Pereira, Alves Rego Cristina Matiele, Ferreira E Ferreira Alex Augusto, Paloschi Mauro Valentino, Soares Andreimar M, Fortes-Dias Consuelo Latorre, Setúbal Sulamita Silva, Zuliani Juliana Pavan
Laboratório de Imunologia Celular Aplicada à Saúde, Fundação Oswaldo Cruz, Fiocruz Rondônia, Porto Velho-RO, Brazil.
Laboratório de Biotecnologia de Proteínas e Compostos Bioativos, Labioprot, Fundação Oswaldo Cruz, Fiocruz Rondônia, Porto Velho-RO, Brazil.
Curr Protein Pept Sci. 2025;26(5):352-364. doi: 10.2174/0113892037317894240926081324.
Neutralizing Factor (CNF) is a γ-type Phospholipase A (PLA) inhibitor present in the blood of snake. Particularly, CNF inhibits the toxic action of Crotoxin (CTX), which is a major neurotoxin found in venom. CTX induces also myotoxic action and demonstrates high selectivity for skeletal muscle fibers. Consequently, CTX can diffuse beyond the site of infection, which can potentially evoke rhabdomyolysis. The present study has evaluated the effects of CTX on myoblasts and myotubes of muscle cells C2C12 and the effect of CNF on CTX-induced damage.
Cytotoxicity assays were performed by measuring the mitochondrial enzyme dehydrogenase levels. Furthermore, creatine kinase and lactate dehydrogenase levels were used as indicators of muscle damage.
Crotoxin has been found to have cytotoxic effects on C2C12 myoblast cells, while CNF has not shown toxic effects on these cells. Furthermore, the findings have shown CNF (50 μg/mL) to abolish CTX toxicity in myoblasts. The myotubes, differentiated cells, showed no change in mitochondrial respiration when exposed to CNF or CTX, showing greater resistance to the toxic actions of crotoxin.
The data have confirmed the potential of CNF as an anti-myotoxic agent to prevent CTX-damaged myoblasts and increase resistance to the toxic effects of crotoxin on differentiated cells.
中和因子（CNF）是一种存在于蛇血液中的γ型磷脂酶A（PLA）抑制剂。特别地，CNF可抑制响尾蛇毒素（CTX）的毒性作用，CTX是蛇毒中一种主要的神经毒素。CTX还会引发肌毒性作用，并且对骨骼肌纤维具有高度选择性。因此，CTX可扩散至感染部位以外，这可能会引发横纹肌溶解。本研究评估了CTX对肌肉细胞C2C12的成肌细胞和肌管的影响，以及CNF对CTX诱导损伤的影响。
通过测量线粒体酶脱氢酶水平进行细胞毒性测定。此外，肌酸激酶和乳酸脱氢酶水平用作肌肉损伤的指标。
已发现响尾蛇毒素对C2C12成肌细胞具有细胞毒性作用，而CNF对这些细胞未显示出毒性作用。此外，研究结果表明CNF（50μg/mL）可消除成肌细胞中的CTX毒性。分化细胞肌管在暴露于CNF或CTX时线粒体呼吸没有变化，显示出对响尾蛇毒素毒性作用的更大抵抗力。
数据证实了CNF作为抗肌毒性剂的潜力，可预防CTX损伤的成肌细胞，并增加对响尾蛇毒素对分化细胞毒性作用的抵抗力。
