ABSTRACT
An attempt has been made in this communication to develop antiserum in rabbit against Scatophagus. argus sting extract. Antiserum did not neutralized the sting extract induced proinflammatory and haemorrhagic activity but successfully neutralized lethality upto 2LD50. Cyproheptadine, indomethacin and BW 755C pretreatment significantly reduced sting extract induced proinflammatory activity. The haemorrhagic activity of sting extract was significantly inhibited by temperature, UV-exposure, EDTA, cyproheptadine, indomethacin and BW 755C pretreatment. The results conclude that the local effects of S.argus venom is likely to be mediated through release of mediators and may be encountered by pharmacological antagonists better than the antiserum.
Subject(s)
Fish Venoms/chemistry , Fish Venoms/pharmacology , Immune Sera/chemistry , 4,5-Dihydro-1-(3-(trifluoromethyl)phenyl)-1H-pyrazol-3-amine/pharmacology , Animals , Cyproheptadine/chemistry , Edema/chemically induced , Edema/pathology , Edetic Acid/chemistry , Edetic Acid/pharmacology , Fish Venoms/immunology , Hemagglutination , Hemorrhage/chemically induced , Indomethacin/pharmacology , Inflammation , Male , Mice , Perciformes , Rabbits , Rats , Temperature , Time Factors , Ultraviolet RaysABSTRACT
A haemorrhagic protein toxin (SA-HT) was isolated and purified from the spine extract of the Indian venomous butterfish, S. argus Linn, by two step ion exchange chromatography. The toxin was homogeneous in native and SDS-PAGE gel. SDS-molecular weight of the toxin was found to be 18.1 +/- 0.09 kDa. SA-HT produced severe haemorrhage on stomach wall but devoid of cutaneous haemorrhage. UV, EDTA, trypsin, protease, cyproheptadine, indomethacin, acetylsalicylic acid and BW755C treatment significantly antagonized the haemorrhagic activity of SA-HT. The toxin produced dose and time dependent oedema on mice hind paw, which was significantly encountered by cyproheptadine, indomethacin and BW755C. SA-HT increased capillary permeability on guinea pig dorsal flank. On isolated guineapig ileum, rat fundus and uterus, SA-HT produced slow contraction which was completely antagonised by prostaglandin blocker SC19220. On isolated rat duodenum, SA-HT produced slow relaxation. SA-HT significantly increased plasma plasmin, serum MDA level and decreased serum SOD level indicating the possible involvement of cyclooxygenase and lipooxygenase pathway.
Subject(s)
Fish Proteins/chemistry , Fish Proteins/isolation & purification , Fish Venoms/chemistry , Fish Venoms/isolation & purification , 4,5-Dihydro-1-(3-(trifluoromethyl)phenyl)-1H-pyrazol-3-amine/pharmacology , Animals , Anti-Inflammatory Agents, Non-Steroidal/pharmacology , Aspirin/pharmacology , Capillaries , Chromatography, Ion Exchange , Cyproheptadine/pharmacology , Dose-Response Relationship, Drug , Edema/chemically induced , Edetic Acid/pharmacology , Electrophoresis, Polyacrylamide Gel , Female , Gastrointestinal Agents/pharmacology , Guinea Pigs , Indomethacin/pharmacology , Lipoxygenase/metabolism , Mice , Muscle, Smooth/drug effects , Perciformes , Permeability , Rats , Spine/metabolism , Superoxide Dismutase/metabolism , Time Factors , Trypsin/pharmacology , Ultraviolet Rays , Uterus/drug effectsABSTRACT
A sting of the fish S. argus, a venomous edible spotted butterfish, produces tremendous local pain, severe swelling, rise of body temperature, throbbing sensation etc. To establish the pharmacological activities of S. argus sting extract, the present investigation, was carried out on experimental animals. The LD50 of extract was found to be 9.3 mg/kg (iv) in male albino mice. The extract showed loss of sensation, urination and salivation in mice. It potentiated pentobarbitone induced sleeping time in male albino mice and produced hypothermia. Extract produced a fall of cat and guinea pig blood pressure, which was completely abolished by mepyramine. It produced a transient reduction of respiratory rate in rat, but decreased respiratory amplitude in cat, which was abolished after vagotomy. On isolated toad heart, the extract increased both the amplitude and rate of contraction. On isolated guinea pig heart, the sting extract decreased both the rate and amplitude of contraction leading to cardiac arrest, but it had no effect on isolated guinea pig auricle. The extract produced a reversible blockade of electrically induced twitch response of isolated chick biventer cervices preparation, but it had no effect on the isolated rat phrenic nerve diaphragm preparation. It produced a slow contractile response on isolated guinea pig ileum, rat uterus and rat fundal strip preparations but produced slow relaxation on isolated rat duodenum preparation. The contractile response on isolated guinea pig ileum and rat fundal strip was antagonised by SC19220. It did not produce any significant cutaneous haemorrhage in mice and did not produce any haemolysis on saline washed erythrocytes. The sting extract significantly increased capillary permeability of guinea pig dorsal flank and produced oedema in mice hind paw.
Subject(s)
Fish Venoms/pharmacology , Animals , Behavior, Animal/drug effects , Blood Pressure/drug effects , Body Temperature/drug effects , Capillaries/pathology , Cats , Chickens , Edema/chemically induced , Female , GABA Modulators , Guinea Pigs , Hypothermia , Ileum/metabolism , Male , Mice , Muscle, Skeletal/metabolism , Muscle, Smooth/drug effects , Myocardial Contraction/drug effects , Pentobarbital/pharmacology , Perciformes , Permeability , Phrenic Nerve/pathology , Ranidae , Rats , Sleep/drug effects , Uterus/metabolismABSTRACT
A lethal cardiotoxic (BO-CT; Bidder's organ cardiotoxin) protein was purified from the Bidder's organ of the common Indian toad B. melanostictus by gel filtration on Sephadex G-200. The homogeneity of cardiotoxin was tested by gel electrophoresis. The molecular weight of lethal BO-CT was 62 KDa and was devoid of glycoprotein. LD50 of the BO-CT was 50 micrograms/20 g (i.v.) in male albino mice. On isolated heart and auricle BO-CT initially increased the rate and amplitude of contraction and finally produced irreversible blockade of contraction. BO-CT induced auricular blockade, was not influenced by verapamil, propranolol and atropine. On isolated chick biventer cervicis preparation BO-CT produced irreversible blockade of electrically induced twitch response followed by contracture. This action was not antagonized by 4-aminopyridine and neostigmine. BO-CT induced contracture on chick biventer cervicis was increased by Ca2+, decreased by Na+ and abolished by K+. Cardiotoxic and neuromuscular activity of BO-CT was heat stable and abolished by proteolytic enzyme.
Subject(s)
Bufonidae/metabolism , Heart/drug effects , Hemolysin Proteins/isolation & purification , Animals , Guinea Pigs , Hemolysin Proteins/pharmacology , Lethal Dose 50 , Male , Mice , Muscle, Skeletal/drug effectsABSTRACT
Isolation and purification of a lethal protein toxin from the Indian catfish Plotosus canius, Hamilton, venom is described. The purification procedure involved ammonium sulfate precipitate of crude venom followed by DEAE-ion exchange chromatography. The purified toxin (toxin-PC) was homogeneous on one-dimensional PAGE and PAS-negative, and had a molecular weight 15 Kd. Toxin-PC was lethal (LD50 225 micrograms/kg, intravenous, in mice) and cardiotoxic, having neuromuscular blocking activity. Toxin-PC produced cardiac arrest on isolated toad and guinea pig hearts. Prior administration of atropine and propanolol failed to counteract toxin activity on isolated heart preparations. On isolated chick biventer cervicis, toxin-PC produced total blockage of electrically-induced twitch response without affecting carbachol- and acetylcholine-induced contraction. The tension developed by the muscle was Ca++ ion-dependent. Neuromuscular blocking time was reduced when K+ ion concentration was increased in the medium. Antiserum raised against toxin-PC failed to antagonize lethal activity of toxin-PC in mice. Toxin-PC probably represents a major toxic component of catfish venom (P. canius), and was responsible for the pathophysiological changes.
