Venom Fractions Research Articles

The peptide fraction of Bothrops jararaca snake venom induces toxicological effects on the male reproductive system after local envenomation in mice

Bothrops envenomation is complex and provokes prominent local tissue damage and systemic disturbances, but little is known about their effects on the male reproductive system. After intratesticular injection, the bioactive peptide fraction (Bj-PF) obtained from Bothrops jararaca snake venom changes the structure of different stages of the seminiferous epithelium cycle in adult mice. For the first time, we investigated whether local envenomation of Bj-PF induces toxicological effects on the male reproductive system, particularly on the seminiferous epithelium and Sertoli cells. Male adult mice were treated with 0.24 mg.kg−1 by intramuscular (i.m.) injection for 24 h. The testes samples were collected for morphological and morphometric evaluation. The toxicological effects of Bj-PF were also analyzed on mitochondrial metabolism and nitrite (NO2) production in 15P-1 Sertoli cell culture. Bj-PF changed the structure and function of the seminiferous epithelium, particularly the disruption of the epithelium and the presence of degenerated germ cells in the adluminal compartment, but there were no alterations in the basal compartment. Bj-PF increased the thickness of the seminiferous epithelium and decreased the lumen diameter of the tubule. Semiquantitative histological assessment of the degree of tubule degeneration revealed that Bj-PF also increased the number of hypospermatogenic tubules compared to control. Bj-PF reduced NO2 levels in 15P-1 Sertoli cells without changing the mitochondrial metabolism. Overall, the fact that Bj-PF alters the structure and function of the seminiferous epithelium suggests that bioactive peptides found in B. jararaca snake venom can have toxicological effects on the reproductive systems of affected male mice, providing new insight into the biological characteristics of snake venom and therapeutic strategies for envenomation inflammation.

Oxilipin, a New Anti-cancer Phospholipase A2-like Protein from Iranian Caspian Cobra, Naja Naja Oxiana.

The discovery of novel anti-cancer agents from natural resources is highly necessary. Concerning the above problem, the purpose of this study was to discover an anti-cancer compound from Caspian cobra venom. Fractionation of Caspian cobra venom was performed by gel filtration and IEX chromatography. The results showed an anti-cancer protein nominated as Oxilipin. Activity and toxicity of Oxilipin were studied on the colon SW480 cancer cell line using MTT, LDH release, PI staining, morphological cell analysis, hemolysis, and anti-proliferation assays. Oxilipin, an 11kDa protein purified from the venom of the Caspian cobra. LC/MS/MS analysis of obtained protein showed homology with Phospholipase A2 from ‎Naja naja oxiana. 40 µg/ml of Oxilipin can induce an apoptotic effect in SW480 cell line up to 90%; meanwhile, this amount can induce only one-third of cytotoxicity on a normal cell. In this study, Iranian cobra venom was found to have cytotoxic effects on SW480 colon cancer tumor cells, with the least cytotoxicity on normal cells on HEK-293. Given that Oxilipin has slight toxicity on normal cells, it can be hypothesized that the obtained peptide can be considered as a drug lead in an animal model study of colon cancer.

Antibodies against a single fraction of Micrurus dumerilii venom neutralize the lethal effect of whole venom

The coralsnake Micrurus dumerilii (Elapidae) is reported to cause envenomings of medical importance. Previous studies characterized the protein composition of its venom, with phospholipase A2 (PLA2) proteins the most abundant. However, it is unknown which venom components are responsible for its lethal toxicity. Fractionation of M. dumerilii venom from Colombia was carried out using RP-HPLC and each fraction was screened for lethal effect in mice at a dose of 20 μg by intraperitoneal route. Results showed that only one fraction, F9, was lethal. This fraction displayed PLA2 activity, induced indirect hemolysis in vitro, as well as edema and myotoxicity in vivo. SDS-PAGE of unreduced F9 evidenced two bands of 8 and 15 kDa, respectively, consistent with the detection of proteins with masses of 13,217.77 Da, 7144.06 Da, and 7665.55 Da. Tryptic digestion of F9 followed by nESI-MS/MS revealed peptide sequences matching proteins of the three-finger toxin (3FTx) and PLA2 families. Immunization of a rabbit with F9 proteins elicited antibody titers up to 1:10,000 by ELISA. After serum fractionation with caprylic acid, the obtained IgG was able to neutralize the lethal effect of the complete venom of M. dumerilii using a challenge of 2 ×LD50 at the IgG/venom ratio of 50:1 (w/w). In conclusion, present results show that the lethal effect of M. dumerilii venom in mice is mainly driven by one fraction which contains 3FTx and PLA2 proteins. The antibodies produced against this fraction cross-recognized other PLA2s and neutralized the lethal effect of whole M. dumerilii venom, pointing out to the potential usefulness of F9 as a relevant antigen for improving current coral snake antivenoms.

Fractionation of Vipera berus berus Snake Venom and Detection of Bioactive Compounds Targeted to Blood Coagulation System

Introduction. The performed research focused on a search for new biologically active compounds acting on blood coagulation system proteins and cells. To achieve this goal, we fractionated Vipera berus berus snake venom and studied the action of the separated fractions on human blood plasma, fibrinogen, platelets or red cells. Methods. Crude venom was fractionated using ion-exchange chromatography. Protein composition of fractions was studied using SDS-PAGE. The ability of fractions to prolong or initiate blood plasma clotting was studied using the prothrombin time test with thromboplastin. Fibrinogen-specific proteases were detected using enzyme-electrophoresis. Action on red cells was estimated using the hemolysis test. Aggregometry was used for the detection of action on platelets. All experiments in this study were performed in vitro. Results. We obtained fractions containing phospholipase and a protease that is able to hydrolyze fibrinogen, leading to the loss of its ability to polymerize and to maintain platelet aggregation. Conclusion. Further purification and study of these components can be a promising research direction for biotechnological as well as for biomedical use.

Comparative study of the analgesic effects of Bungarus fasciatus snake venom from Vinh Phuc and Tien Giang Provinces of Vietnam

Purpose: To determine the analgesic activity of Bungarus fasciatus venoms and their fractions from two Vietnamese Provinces.
 Methods: Male Swiss Albino mice were randomly divided into three groups containing 8 to 10 mice each. Control group was injected subcutaneously with normal saline, standard group received aspirin solution (50 mg/kg) perorally, and study group received a solution of crude venom or isolated fractions in physiological saline. To determine analgesic activity, acetic acid writhing and tail immersion tests were used. The venoms were separated by liquid chromatography and the analgesic activity of the fractions was analyzed.
 Results: Both venoms showed analgesic effect in the acetic acid writhing test, but only the venom from Tien Giang showed analgesic effect in the tail immersion test. The bioactive fractions of Vinh Phuc and Tien Giang venoms were significantly different, with most of Vinh Phuc venom fractions being more active (p < 0.05). Thus, 35 min after the injection, the number of writhings decreased from 15 - 16 in the control to 0.85 ± 0.34 for the BF-4VS (Vinh Phuc) fraction compared to 2.67 ± 1.20 (p < 0.05) for the BF-4DT (Thien Giang) fraction. Two proteins with analgesic activity were isolated from Vinh Phuc venom, and one with greater activity matched the known B. fasciatus phospholipase A2.
 Conclusion: The analgesic activity of two samples of B. fasciatus venom from two different provinces in Vietnam reveal that their pharmacological profiles differ. The isolates can be explored as leads in the development of new analgesic agents.

Purification and Characterization of a Novel Factor of Crotoxin Inter-CRO (V-1), a New Phospholipase A2 Isoform from Crotalus durissus collilineatus Snake Venom Using an In Vitro Neuromuscular Preparation

The fractionation of Crotalus durissus collilineatus whole venom through an HPLC chromatographic method enabled the purification of a new V-1 neurotoxin. Inter-CRO (V-1) presents similarity in its primary structure to crotoxin B (CB), suggesting another isoform of this toxin. The aim of this study was to compare V-1 to the crotoxin complex (CA/CB) and CB to elucidate aspects related to its functionality. The homogeneity of the purified protein was confirmed with a molecular mass of 1425.45 Da, further verified by mass spectrometry. The sequence of the protein showed high similarity to other viperid snake venom PLA2 proteins. The results of this study report that V-1 is an uncharacterized novel toxin with different biological activities from CB. V-1 maintained catalytic activity but presented neurotoxic activity as observed by the 2.5-fold increase in twitch tension record compared to control values on isolated muscle cells.

Origins, genomic structure and copy number variation of snake venom myotoxins

Crotamine, myotoxin a and homologs are short peptides that often comprise major fractions of rattlesnake venoms and have been extensively studied for their bioactive properties. These toxins are thought to be important for rapidly immobilizing mammalian prey and are implicated in serious, and sometimes fatal, responses to envenomation in humans. While high quality reference genomes for multiple venomous snakes are available, the loci that encode myotoxins have not been successfully assembled in any existing genome assembly. Here, we integrate new and existing genomic and transcriptomic data from the Prairie Rattlesnake (Crotalus viridis viridis) to reconstruct, characterize, and infer the chromosomal locations of myotoxin-encoding loci. We integrate long-read transcriptomics (Pacific Bioscience's Iso-Seq) and short-read RNA-seq to infer gene sequence diversity and characterize patterns of myotoxin and paralogous β-defensin expression across multiple tissues. We also identify two long non-coding RNA sequences which both encode functional myotoxins, demonstrating a newly discovered source of venom coding sequence diversity. We also integrate long-range mate-pair chromatin contact data and linked-read sequencing to infer the structure and chromosomal locations of the three myotoxin-like loci. Further, we conclude that the venom-associated myotoxin is located on chromosome 1 and is adjacent to non-venom paralogs. Consistent with this locus contributing to venom composition, we find evidence that the promoter of this gene is selectively open in venom gland tissue and contains transcription factor binding sites implicated in broad trans-regulatory pathways that regulate snake venoms. This study provides the best genomic reconstruction of myotoxin loci to date and raises questions about the physiological roles and interplay between myotoxin and related genes, as well as the genomic origins of snake venom variation.

Venom fractions of Philippine tarantula species exhibits antiangiogenic activity in ovo

Venom fractions of Philippine tarantula species exhibits antiangiogenic activity in ovo

Profilings of subproteomes of lectin-binding proteins of nine Bothrops venoms reveal variability driven by different glycan types

Snake venom proteomes have long been investigated to explore a multitude of biologically active components that are used for prey capture and defense, and are involved in the pathological effects observed upon mammalian envenomation. Glycosylation is a major protein post-translational modification in venoms and contributes to the diversification of proteomes. We have shown that Bothrops venoms are markedly defined by their content of glycoproteins, and that most N-glycan structures of eight Bothrops venoms contain sialic acid, while bisected N-acetylglucosamine was identified in Bothrops cotiara venom. To further investigate the mechanisms involved in the generation of different venoms by related snakes, here the glycoproteomes of nine Bothrops venoms (Bothrops atrox, B. cotiara, Bothrops erythromelas, Bothrops fonsecai, B. insularis, Bothrops jararaca, Bothrops jararacussu, Bothrops moojeni and Bothrops neuwiedi) were comparatively analyzed by enrichment with three lectins of different specificities, recognizing bisecting N-acetylglucosamine- and sialic acid-containing glycoproteins, and mass spectrometry. The lectin capture strategy generated venom fractions enriched with several glycoproteins, including metalloprotease, serine protease, and L- amino acid oxidase, in addition to various types of low abundant enzymes. The different contents of lectin-enriched proteins underscore novel aspects of the variability of the glycoprotein subproteomes of Bothrops venoms and point to the role of distinct types of glycan chains in generating different venoms by closely related snake species.

ACTION OF VENOM OF VIPERA SNAKE OF UKRAINE ON BLOOD COAGULATION in vitro

Aim. In this study we focused on the search of fibrinogen-targeted proteases in the venom of Vipera renardi, Vipera nikolskii and Vipera berus. Venom of Vipera berus was also fractionated on Q-sepharose and action of separated fractions on human blood plasma, platelets and red cells was studied. Methods. Analysis of protein mixtures was performed using SDS-PAGE. Аction on the blood coagulation system was analyzed using the APTT assay. Identification of protein components with fibrinolytic activity was performed using enzyme-electrophoresis with fibrinogen as the substrate. Fractionation of V. berus venom was performed on Q-sepharose using FPLC system Acta Prime. Action of separated fractions on ADP-induced platelet aggregation in platelet rich blood plasma was analyzed using Aggregometer AP 2110. Hemolytic action of fractions was estimated using fresh human red cells. Amount of released hemoglobin was estimated by spectrophotometry on Optizen POP. Results. All studied venoms had different protein compositions with major protein fractions in the range from 25 kDa to 130 kDa. Both V. berus and V. nikolskii venoms taken in 1:200 dilutions reduced the time of clotting in APTT test from 25 to 13 s. In contrast, V. renardi venom in the same dilution prolonged the clotting time from 25 s to 180 s that we assumed as the result of fibrinogen-specific protease presence. According to enzyme-electrophoresis data all studied venoms contained fibrinogen-specific proteases with the apparent molecular weights for V. berus, V. nikolskii – 25-55 kDa. and V. renardi – 55-75 kDa. Fractionation of crude venom of V. berus allowed obtaining several fractions eluted at different concentrations of NaCl: 0.1; 0.2; 0.3; 0.5 М. Non-binded fraction was also collected. Conclusions. Thus, the components of Vipera venoms living in Ukraine can be used for basic biochemical research. At the same time, care should be taken in the case of envenomation, as the presence of fibrinogenolytic enzymes in the venom can lead to hemorrhage. Further characterization of fibrinogen-specific protease from V. berus venom is a promising task for biotechnology.

In-Vitro Evaluation of Anticancer and Antibacterial Properties of Pseudocerastes Persicus Snake Venom Fractions

Background and Objectives In recent years the isolation of effective molecules from snake venom as drug to treatment of some incurable diseases is considered by many biological research centers. The aim of this investigation was to determine electrophorotic and chromatographic patterns of Pseudocerastes persicus venom and in-vitro study of bactericidal and anti-cancer properties of its fractions in human hepatocellular liver carcinoma cell line (HepG2). Subjects and Methods The fractions of venom were isolated and collected using preparative RP-HPLC. The Bactericidal activity of fractions in 20μg/ml of protein concentration was investigated toward Gramnegative and Gram-positive bacteria using MTT and MIC assays. Then, the cytotoxic effects of fractions with higher antibacterial properties were measured by MTT reduction, neutral uptake and comet assays following exposure of HepG2 cancer cells to 20 and 40 μg/ml of protein concentration. 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 separation of fractions from the venom by HPLC resulted in collection of 11 different fractions. The results of this study showed that two fractions (4 & 8) have a significant antibacterial activity against Gram-positive as compared to Gram- negative bacteria. The study of the anti-cancer effect of these two fractions also showed that fraction 8 was more toxic in HepG2 cells. Conclusion We for the first time reported bactericidal and anti cancer 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 anti tumor activities.

Evaluation of in vivo Lethality and in vitro Cytotoxic Effect of Odontobuthus bidentatus Scorpion Venom.

The results of numerous studies have revealed that some deadly scorpion venoms are composed of various bioactive molecules that have significant cytotoxic effects on cancer cells. In this study, the in vivo lethality and cytotoxic effect of Odontobuthus bidentatus venom were evaluated in different cancer cell lines. Through MTT assay, the cytotoxic effects of O. bidentatus scorpion venom were analyzed on the MCF-7, A549, AGS, HepG2, and Ht-29 cancer cell lines and Hu02 normal cells. To this end, six venom fractions were obtained through a Sephadex G-50 column, and the cytotoxic effects of isolated fractions were evaluated on A549 lung cancer cells. The median lethal dose of O. bidentatus scorpion venom was determined at 0.73 mg/kg by intravenous administration of different venom doses in male BALB/c mice according to the Spearman-Karber method. The O. bidentatus scorpion whole venom had a significant cytotoxic effect on MCF-7, A549, and AGS cells. The treatment of A549 cells with various concentrations of fraction F1 showed that this fraction significantly induced growth inhibitory effect on the cells in a dose-dependent manner, compared to untreated cells.

Local envenomation caused by a bioactive peptide fraction of Bothrops jararaca snake venom induces leukocyte influx in the lung and changes in pulmonary mechanics

The crude venom of the Bothrops jararaca snake (Bj-CV) is a complex mixture of biologically active proteins that includes a variety of peptides in the low molecular weight fraction (Bj-PF). We investigated how an intramuscular injection of Bj-CV (1.2 mg kg−1) and Bj-PF (0.24 mg kg−1) influenced lung mechanics and lung and muscle inflammation in male Swiss mice 15 min, 1, 6, and 24 h after inoculation. Pressure dissipation against lung resistive components (ΔP1) rose significantly from 1 to 24 h after Bj-CV and 6–24 h after Bj-PF inoculation. Both Bj-CV and Bj-PF increased the total pressure variation of the lung (ΔPtot) 24 h after injection. Lung static elastance increased significantly after injection in all time periods investigated by Bj-CV and from 6 to 24 h by Bj-PF. Lung static elastance increased significantly after injection in all time periods investigated by Bj-CV and from 6 to 24 h by Bj-PF. Furthermore, intramuscular inoculation of Bj-CV and Bj-PF resulted in an increase in muscle and pulmonary inflammation, as evidenced by an increase in leukocyte influx when compared to the control group. Finally, both Bj-CV and Bj-PF cause acute lung injury, as shown by pulmonary inflammation and decreased lung mechanics. Furthermore, the fact that Bj-PF produces mechanical alterations in the lungs and muscular inflammation implies that non-enzymatic compounds can cause inflammation.

Inhibitory Effect of Hemiscorpius lepturus Scorpion Venom Fractions on Tachyzoites of Toxoplasma gondii.

Background:The present study determined the effect of the fractions obtained from Hemiscorpius lepturus scorpion venom on the tachyzoite of Toxoplasma gondii.Methods:The fractions of dried venom of He. lepturus scorpion of Khuzestan Province, southern Iran in 2019 were isolated through gel filtration chromatography, and then tachyzoites were exposed to fractions of venom at different concentrations. Trypan blue counting and MTT were applied to assay tachyzoite viability, and the inhibition of the cellular growth of fractions in Vero cells was evaluated.Results:The maximum effect on tachyzoite was observed in fraction 5 of venom. To further separate the protein, fraction 5 was used in high-performance liquid chromatography assay to purify its proteins. Based on the results of HPLC of fraction 5, among which the second peak, a peptide with <10 KDa representing a more potent effect in eliminating the tachyzoite of T. gondii.Conclusion:The scorpion venom-purified fractions possess anti-parasitic activity against the tachyzoite of T. gondii and can be used in parasite-controlling studies.

Clinical cardiac alterations and hemostatic toxicities caused by scorpion (Tityus discrepans) venom and its purified fractions on zebrafish (Danio rerio) larvae

Envenomation by the Venezuelan scorpion Tityus discrepans is typified by local and systemic alterations. The current work investigated the in vivo hemostatic processes, cardiac dysfunction and tissue destruction triggered by Tityus discrepans purified toxins 1 (3 kDa) and 2 (5 kDa) fractions. These fractions were obtained by C-18-HPLC chromatography. The hemostatic and cardiovascular toxicities in zebrafish of both fractions was assessed by means of specific phenotypic expressions and larvae behavior at 5, 15, 30, 40 and 60 min post-venom-treatment. The Tityus discrepans venom fractions 1 and 2 produced disseminated intravascular coagulation (presence of thrombus) in the central vein of the larva, heart rate/rhythm alterations, and necrotic events in more than 90% of all the larvae under their action. The outcomes have established the potential hemostatic and cardiovascular toxicities by Tityus discrepans venom, alerting on the possibility of cardiovascular injuries and thromboembolism in humans after scorpion stings envenomation.

Bee Venom: From Venom to Drug.

Insects of the order Hymenoptera have a defensive substance that contains many biologically active compounds. Specifically, venom from honeybees (Apis mellifera) contains many enzymes and peptides that are effective against various diseases. Different research papers stated the possibility of using bee venom (a direct bee sting or in an injectable form) in treating several complications; either in vivo or in vitro. Other reports used the active fractions of bee venom clinically or at labratory scale. Many reports and publications have stated that bee venom and its constituents have multiple biological activities including anti-microbial, anti-protozoan, anti-cancer, anti-inflammatory, and anti-arthritic properties. The present review aims to refer to the use of bee venom itself or its fractions in treating several diseases and counteracting drug toxicities as an alternative protocol of therapy. The updated molecular mechanisms of actions of bee venom and its components are discussed in light of the previous updated publications. The review also summarizes the potential of venom loaded on nanoparticles as a drug delivery vehicle and its molecular mechanisms. Finally, the products of bee venom available in markets are also demonstrated.

The Metabolomic Profiles of Sera of Mice Infected with Plasmodium berghei and Treated by Effective Fraction of Naja naja oxiana Using 1H Nuclear Magnetic Resonance Spectroscopy.

The use of venom fractions from the Iranian cobra could be useful adjunct treatments of malaria with chloroquine. A metabolomic investigation with 1HNMR spectroscopy was conducted on an effective fraction tested earlier using Plasmodium berghei as an experimental murine model. We sought to ascertain both safety and anti-parasitic effects of experimental therapies. After purification of the venom fractions, 25 mice were infected, then treated for 4days with 0.2ml of 5mg/kg, 2.5mg/kg and 1mg/kg of the effective fraction, chloroquine, and a drug vehicle. An ED50 was obtained using Giemsa staining and real-time PCR analysis. The toxicity tests inspecting both liver and kidney tissues were performed. A clear inhibitory effect on parasitaemia was observed (with 75% inhibition with 5mg/kg and 50% reduction when 2.5mg/kg dosage used). ED50 obtained 2.5 mg/kg. The metabolomics were identified as differentiation of aminoacyl-t-RNA biosynthesis, valine, leucine, isoleucine biosynthesis and degradation pathways were observed. Upon therapeutic effects of cobra venom fraction, further optimization of dose-dependent response of pharmacokinetics would be worthwhile for further exploration in adjunct experimental venom therapies.

Evaluation of in-vitro bactericidal activity and anticancer effects of venom fractions of Pseudocerastes persicus snake

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.

Synergism of in vitro plasmodicidal activity of phospholipase A2 isoforms isolated from panamanian Bothrops asper venom

Bothrops asper is one of the most important snake species in Central America, mainly because of its medical importance in countries like Ecuador, Panama and Costa Rica, where this species causes a high number of snakebite accidents. Several basic phospholipases A2 (PLA2s) have been previously characterized from B. asper venom, but few studies have been carried out with its acidic isoforms. In addition, since snake venom is a rich source of bioactive substances, it is necessary to investigate the biotechnological potential of its components. In this context, this study aimed to carry out the biochemical characterization of PLA2 isoforms isolated from B. asper venom and to evaluate the antiparasitic potential of these toxins. The venom and key fractions were subjected to different chromatographic steps, obtaining nine PLA2s, four acidic ones (BaspAc-I, BaspAc-II, BaspAc-III and BaspAc-IV) and five basic ones (BaspB-I, BaspB-II, BaspB-III, BaspB-IV and BaspB-V). The isoelectric points of the acidic PLA2s were also determined, which presented values ranging between 4.5 and 5. The findings indicated the isolation of five unpublished isoforms, four Asp49-PLA, corresponding to the group of acidic isoforms, and one Lys49-PLA2-like. Acidic PLA2s catalyzed the degradation of all substrates evaluated; however, for the basic PLA2s, there was a preference for phosphatidylglycerol and phosphatidic acid. The antiparasitic potential of the toxins was evaluated, and the acidic PLA2s demonstrated action against the epimastigote forms of T. cruzi and promastigote forms of L. infantum, while the basic PLA2s BaspB-II and BaspB-IV showed activity against P. falciparum. The results indicated an increase of up to 10 times in antiplasmodial activity, when the Asp49-PLA2 and Lys49-PLA2 were associated with one another, denoting synergistic action between these PLA2 isoforms. These findings correspond to the first report of synergistic antiplasmodial action for svPLA2s, demonstrating that these molecules may be important targets in the search for new antiparasitic agents.

Anti-Cancer Effect of Moroccan Cobra Naja haje Venom and Its Fractions against Hepatocellular Carcinoma in 3D Cell Culture.

Hepatocellular carcinoma (HCC) is the most common primary liver cancer in adults, the fifth most common malignancy worldwide and the third leading cause of cancer related death. An alternative to the surgical treatments and drugs, such as sorafenib, commonly used in medicine is necessary to overcome this public health problem. In this study, we determine the anticancer effect on HCC of Moroccan cobra Naja haje venom and its fraction obtained by gel filtration chromatography against Huh7.5 cancer cell line. Cells were grown together with WI38 human fibroblast cells, LX2 human hepatic stellate cell line, and human endothelial cells (HUVEC) in MCTS (multi-cellular tumor spheroids) models. The hepatotoxicity of venom and its fractions were also evaluated using the normal hepatocytes cell line (Fa2N-4 cells). Our results showed that an anti HCC activity of Moroccan cobra Naja haje venom and, more specifically, the F7 fraction of gel filtration chromatography exhibited the greatest anti-hepatocellular carcinoma effect by decreasing the size of MCTS. This effect is associated with a low toxicity against normal hepatocytes. These results strongly suggest that the F7 fraction of Moroccan cobra Naja haje venom obtained by gel filtration chromatography possesses the ability to inhibit cancer cells proliferation. More research is needed to identify the specific molecule(s) responsible for the anticancer effect and investigate their mechanism of action.