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 SDS-PAGE 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.
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