Introduction: It was reported that snake venom contains a wide mixture of proteins and peptides with various toxicological and pharmacological effects. In recent years, the isolation of effective molecules from snake venom as a drug to treatment of some incurable diseases is considered by many biological research centers. This investigation aimed to determine electrophoretic and chromatographic patterns of Pseudocerastes persicus snake venom and in-vitro study of bactericidal and anti-cancer properties of its fractions in a human hepatocellular liver carcinoma cell line (HepG2). Materials and methods: The fractions of venom were isolated and collected using preparative RP-HPLC. The Bactericidal activity of fractions in 20 μg/ml of protein concentration was investigated toward Gram-negative and Gram-positive bacteria. Antibacterial susceptibility tests were done using MTT and MIC assays. Furthermore, cell viability was measured by MTT reduction assay and confirmed with Neutral red uptake assay following exposure of HepG2 cancer cells to 20 and 40 μg/ml of protein concentrations of fractions having more antibacterial properties. Apoptotic effects were investigated using the comet assay. Results: SDS-PAGE pattern of the crude venom revealed 10 major bands with molecular weight ranging from 13.2 to 99.25 kDa and the single-step separation of different fractions from the venom by HPLC resulted in isolation and collection of 11 fractions. The results of this study showed that two fractions of snake venom have significant antibacterial activity against Gram-positive as compared to Gram-negative bacteria. On the other hand, our findings indicated that one of them is not toxic to the HepG2 cells. Discussion: We for the first time reported bactericidal and anticancer effects of isolated fractions of Persian horned viper venom. These properties make components of these fractions a favorable source for isolation of molecule(s) with antibacterial and antitumor activities.
Antibiotic resistance has been reported as one of the world's most critical public health problems. Recent investigations have demonstrated that the venom of some species of snakes has antimicrobial and anticancer activities. The purpose of this study was to examine the antibacterial activity of Persian horned viper venom toward Escherichia coli, Bacillus subtilis, and Staphylococcus aureus bacteria and also investigate antitumor effects of concentrations of venom with higher antibacterial activity in a human hepatocellular liver carcinoma cell line (HepG2) to evaluate the potential use of its components as antimicrobial and anticancer agents. Bactericidal activity of crude venom in concentrations of 6.25-400 μg/ml was performed using MTT reduction, minimum inhibitory concentration (MIC), agar- well diffusion, and disc diffusion methods. Tetracycline (50 μg/ml) was used as a standard antibiotic. Cytotoxic effect in HepG2 cells was measured by MTT reduction assay and confirmed with neutral uptake assay following exposure of cells with different concentrations of venom (50-400 μg/ml). The apoptotic effect was investigated using the comet assay. Our findings demonstrated that venom displays higher inhibitory effects against Gram-positive bacteria as compared to Gram- negative. Furthermore, venom showed anticancer activity on the HepG2 cell line through induction of apoptosis and necrosis. The results of this study confirmed that the venom of Persian horned viper induces antibacterial and anticancer effects. These properties make the venom of this viper a potential source for isolation of effective molecule(s) having antibacterial and antitumor activity.
