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Enzymatic analysis of venom from Cuban scorpion Rhopalurus junceus. 古巴蝎子 Rhopalurus junceus 毒液的酶分析。
Pub Date : 2015-07-22 eCollection Date: 2015-01-01
Alexis Díaz-García, Jenny Laura Ruiz-Fuentes, Arianna Yglesias-Rivera, Hermis Rodríguez-Sánchez, Yanelis Riquenes Garlobo, Osmel Fleitas Martinez, José A Fraga Castro

Rhopalurus junceus scorpion venom has been identified as a natural extract with anticancer potential. Interestingly, this scorpion venom does not cause adverse symptoms in humans. However, there is scarce information about its composition and enzymatic activity. In this work, we determined the electrophoretic profile of the venom, the gelatinase and caseinolytic activity, and the phospholipase A2 (PLA2) and hemolytic activity. The effect of different venom doses (6.25, 12.5 and 25 mg/kg) on gastrocnemius muscle was also measured as CK and LDH activity in serum. The presence of hyaluronidase was determined by turbidimetric assay. The effect of different fractions obtained by gel filtration chromatography were evaluated at different concentrations (0.05, 0.1, 0.2, 0.4, 0.6mg/ml) against lung cancer cell A549 and lung normal cell MRC-5 using MTT assay. The electrophoretic profile demonstrated the presence of proteins bands around 67kDa, 43kDa, 18.4kDa and a majority band below 14.3kDa. The venom did not showed caseinolytic, gelatinase, PLA2 and hemolytic activity even at highest venom concentration used in the study. Scorpion venom only showed a significant toxic effect on gastrocnemius muscles identified by CK and LDH release after subcutaneous injection of 12.5 and 25mg/kg. Low molecular weight fractions (<4kDa) induced a significant cytotoxicity in A549 cells while high molecular weight proteins (45-60kDa) were responsible for hyaluronidase activity and toxic effect against MRC-5. Experiments indicate that Rhopalurus junceus scorpion venom has low enzymatic activity, which could contribute to the low toxic potential of this scorpion venom.

君子兰蝎毒已被确认为一种具有抗癌潜力的天然提取物。有趣的是,这种蝎毒不会对人类造成不良症状。然而,有关其成分和酶活性的信息却很少。在这项工作中,我们测定了毒液的电泳图谱、明胶酶和酪蛋白溶解活性、磷脂酶 A2(PLA2)和溶血活性。不同毒液剂量(6.25、12.5 和 25 毫克/千克)对腓肠肌的影响还通过血清中的 CK 和 LDH 活性进行了测定。透明质酸酶的存在是通过比浊法测定的。用 MTT 法评估了凝胶过滤色谱法获得的不同馏分在不同浓度(0.05、0.1、0.2、0.4、0.6 毫克/毫升)下对肺癌细胞 A549 和肺正常细胞 MRC-5 的影响。电泳图谱显示存在 67kDa、43kDa、18.4kDa 左右的蛋白质条带,大部分条带低于 14.3kDa。即使在研究中使用的最高毒液浓度下,蝎毒也没有显示出酪蛋白溶解、明胶酶、PLA2 和溶血活性。皮下注射 12.5 毫克/千克和 25 毫克/千克蝎毒后,蝎毒仅对腓肠肌显示出明显的毒性作用,并通过 CK 和 LDH 释放来确定。低分子量馏分
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Journal of venom research
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