Identification and Purification of the Three Toxins from Odotobuthus bidentatus, Lourenco and Pezier 2002, Venom (Scorpions, Buthidae)

Masihipour, Behzad; Zare, Abbas; Navidpour, Shahrokh; Moghadam, Ahmad Taghavi; Akbari, Abolfazl; Rabiei, Hamed

doi:10.3923/jbs.2009.494.498

Cited by 2 publications

(3 citation statements)

References 13 publications

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“…Venom purification has already been performed in Iran for identification of toxic fractions of some scorpion venoms including Odotobuthus bidentatus and M. eupeus (6,15). Previously, Kadkhodaei et al (6) isolated toxic fractions of M. eupeus venom through two-step purification by gel filtration chromatography with Sephadex G-50 and cation-exchange chromatography.…”

Section: Discussionmentioning

confidence: 99%

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Comparison of two purified toxic fractions from Mesobuthus eupeus scorpion venom

Shirmardi¹,

Shamsaei²,

Ghanei³

et al. 2010

J. Venom. Anim. Toxins incl. Trop. Dis

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Abstract:Iranian scorpions belong mainly to the Buthidae and Scorpionidae families, distributed into 16 genera and 25 species. In Iran, similar to other parts of the world, there are a few known species of scorpions responsible for severe envenoming; amongst which Mesobuthus eupeus is the most common. Its venom contains several toxin fractions that may affect the ion channel. In the present study purification, labeling and biological evaluation of M. eupeus venom are described. For separation, soluble venom was loaded on a chromatography column packed with Sephadex G-50 gel. Subsequently, the fractions were collected according to UV absorption at a wavelength of 280 nm. Toxic fraction (F3) was loaded on an anionic ion exchanger resin and then on a cationic resin. Finally, toxic subfractions F3.1.6 and F3.1.9 were labeled with 99m Tc and injected into normal mice to distinguish excretion pathway. The venom toxic fraction was successfully obtained in its purified form. Radiolabeling of toxic fractions was performed at high specific activity with radiochemical purity of more than 97 and 95% respectively for F3.1.6 and F3.1.9. Biodistribution studies in normal mice with two toxic fractions usually show rapid clearance of the compounds from blood and tissue except for kidneys. Since tissue distribution studies are very important for clinical purpose, the present findings suggest that 99m Tc labeling of venom is a useful tool for in vivo studies and comprises an excellent approach to monitoring the process of biodistribution and kinetics of toxins.

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Section: Discussionmentioning

confidence: 99%

“…The toxic purified fractions of F3.1.6 and F3.1.9 were labeled with 99m Tc using stannous chloride and sodium borohydride as reducing agents (14)(15)(16). Preliminary studies were performed to establish the optimum conditions for obtaining the highest yield of labeled venom.…”

Section: Labeling With 99m Tcmentioning

confidence: 99%

Comparison of two purified toxic fractions from Mesobuthus eupeus scorpion venom

Shirmardi¹,

Shamsaei²,

Ghanei³

et al. 2010

J. Venom. Anim. Toxins incl. Trop. Dis

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“…Four fractions were obtained after passing the venom through the Sephadex G50 column, and it was observed that only fraction 2 caused death in mice. Subsequently, fraction 2 was separated using DEAE Sephadex chromatography, resulting in the isolation of four fractions, of which fractions 1 and 3 exhibited toxicity in mice (42).…”

Section: Discussionmentioning

confidence: 99%

Low molecular weight peptides derived from Iranian Scorpion (Odontobuthus bidentatus) Venom Induces Apoptosis in the Hepatocellular Carcinoma Cell Line (HepG2) in 3D Cell Culture

Alikhani

Zargan

Bidmeshkipour

et al. 2023

Preprint

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Background Hepatocellular carcinoma (HCC) is a deadly form of liver cancer with limited treatment options. Natural compounds, such as venom-derived peptides, have emerged as potential sources of anticancer agents. 3D cell models, which closely resemble the architectural characteristics of natural tumors, serve as an appropriate system for investigating the cytotoxic and apoptotic effects of scorpion venom on cancer cells. In this study, we investigated the apoptotic effects of low molecular weight peptides isolated from the venom of Odontobuthus bidentatus on HepG2 cells in a 3D cell culture model. Methods and Results The O. bidentatus venom was subjected to high-performance liquid chromatography (HPLC) for fractionation and purification of the low molecular weight peptides. Subsequently, the isolated peptides were evaluated for their impact on cell viability and apoptosis induction in HepG2 cells within a 3D cell culture system and were compared to crude venom. To create a 3D cell culture, HepG2 cells were enclosed within alginate hydrogel. Subsequently, the cytotoxic effects of scorpion venom were evaluated using MTT and neutral red uptake assays. Changes in the redox potential of HepG2 cells were assessed by measuring accumulated nitric oxide (NO) in the cell culture media, as well as levels of glutathione (GSH) and catalase activity. To determine the induction of apoptosis in cells treated with scorpion venom, various assays including alkaline comet assay, caspase-3 enzyme activity, and cytochrome c release were employed. Additionally, the expression of the pro-apoptotic gene BAX and the anti-apoptotic gene BCL-2 was evaluated using qRT-PCR. The results obtained from the MTT and neutral red uptake assays demonstrated that O. bidentatus crude venom and isolated fractions (5, 6, and 10) had cytotoxic effects on HepG2 cells in the 3D cell culture. The concentration of NO released into the culture media increased, while the levels of reduced glutathione and catalase decreased in a dose-dependent manner within the 3D culture. The findings from the caspase-3 enzyme activity, cytochrome c release assay, comet assay, and Bax/Bcl-2 gene expression ratio supported the conclusion that O. bidentatus scorpion venom induces apoptosis through the mitochondrial pathway. Conclusion This finding highlights the potential of scorpion venom-derived peptides as novel therapeutic agents for hepatocellular carcinoma.

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Identification and Purification of the Three Toxins from Odotobuthus bidentatus, Lourenco and Pezier 2002, Venom (Scorpions, Buthidae)

Cited by 2 publications

References 13 publications

Comparison of two purified toxic fractions from Mesobuthus eupeus scorpion venom

Comparison of two purified toxic fractions from Mesobuthus eupeus scorpion venom

Low molecular weight peptides derived from Iranian Scorpion (Odontobuthus bidentatus) Venom Induces Apoptosis in the Hepatocellular Carcinoma Cell Line (HepG2) in 3D Cell Culture

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