Effect of frequency of Rhopalurus junceus scorpion venom collection on protein content and biological activity

A. Díaz-García, Arianna Yglesias-Rivera, Jenny L. Ruiz-Fuentes, Rayner Ochoa-Cardentey, Jose Raúl Torres Viltres, Hermis Rodríguez-Sánchez, Tania Gómez Peña
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引用次数: 2

Abstract

Frequency of scorpion venom extraction could influence the quality of venom and as a result may vary their biological activity, whether pharmacological or toxicological behavior, due to venom regeneration is an asynchronous process. In the present study we evaluate the protein content, concentration and biological activity of Rhopalurus junceus scorpion varying the extended periods of venom collection. Three scorpion groups were re-milked at 7, 15 and 21 days after first extraction. Differences in protein content were analyzed by SDSPAGE and HPLC techniques. The biological effect against A549, C6/36HT and Vero cells was assayed by MTT using different venom concentration (0.062; 0.125; 0.25; 0.5; 1 mg/mL). Venom from day 7 and 15 showed significant differences in protein concentration (p<0.05) respect to first extraction. SDS-PAGE and HPLC showed the highest and lowest variation in venom content, at day 7 and 21 respectively. Vero normal cells viability was not affected at all extraction periods. Meanwhile, insect cells, C6/36HT was sensitive at all extraction periods. Extended periods of venom collection was positively correlated with the regeneration of venom composition and the increase of cytotoxic effect against A549 cancer cells. R. junceus venom regeneration showed to be asynchronous depending of the target cell. Besides, venom seems to be completely renewal after 21 days showing high protein content and reproducible results against cancer cells respect to former extraction. *Correspondence to: Alexis Díaz-García, Millennium Nucleus of Ion ChannelsAssociated Diseases (MiNICAD), University of Chile, Faculty of Medicine, North Campus, Ave. Independencia 1027 Hall B, First Floor, Chile, E-mail: alexisdg76@gmail.com
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采集蛇毒次数对蛋白质含量及生物活性的影响
蝎子毒液的提取频率会影响毒液的质量,从而可能改变其生物活性,无论是药理学还是毒理学行为,因为毒液再生是一个异步过程。在本研究中,我们评价了不同时间的毒蝎的蛋白质含量、浓度和生物活性。三组蝎子分别在第一次提取后的第7、15和21天再次挤奶。采用SDSPAGE和HPLC技术分析蛋白质含量差异。采用MTT法测定不同毒液浓度(0.062;0.125;0.25;0.5;1毫克/毫升)。与第一次提取相比,第7天和第15天提取的毒液蛋白质浓度差异显著(p<0.05)。SDS-PAGE和HPLC分别在第7天和第21天显示出毒液含量的最高和最低变化。在所有提取期,Vero正常细胞的活力均未受到影响。同时,昆虫细胞C6/36HT在各提取时段均敏感。延长毒液采集时间与毒液成分的再生和对A549癌细胞的细胞毒作用增强呈正相关。蛇毒的再生是不同步的,取决于不同的靶细胞。此外,毒液在21天后似乎完全更新,与以前的提取相比,蛋白质含量高,对癌细胞的杀伤效果可重复。*通信:亚历克西斯Díaz-García,离子通道相关疾病千年核(MiNICAD),智利大学医学院,北校区,独立大道1027号B厅,智利,一楼,E-mail: alexisdg76@gmail.com
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