Effects of Russell's viper venom fractions on systemic and renal hemodynamics

doi:10.1016/j.toxicon.2006.09.013

Toxicon

Volume 49, Issue 1, January 2007, Pages 82-88

https://doi.org/10.1016/j.toxicon.2006.09.013 Get rights and content

Abstract

Changes in systemic and renal hemodynamics induced by Russell's viper venom are well established. The component of the venom responsible for hemodynamic alteration has not been identified. By Sephadex column chromatography five fractions of Russell's viper (Daboia russellii siamensis) venom were isolated. Each venom fraction consisted of phospholipase A₂, proteolytic enzyme, phosphomonoesterase, phosphodiesterase, arginine ester hydrolase and hyaluronidase of varying activities. Hemodynamic effects of each venom fraction including mean arterial pressure, cardiac output, systemic and renal vascular resistance, renal blood flow and glomerular filtration rate were studied in five groups of dogs; each group had four dogs. Minimal hemodynamic changes were observed in dogs receiving venom fraction I. Increased renal vascular resistance with diminution of renal blood flow and glomerular filtration rate was observed in dogs receiving venom fractions II, III, IV and V. A markedly increased renal vascular resistance with maximal decrease in renal blood flow and glomerular filtration rate was caused by fraction III of the venom with highest PLA₂ and proteolytic enzyme activities. However, renal hemodynamic changes appeared to correlate better with proteolytic enzyme activity than PLA₂ activity. The findings suggested the proteolytic enzyme as an important determinant of hemodynamic alteration. Fractional excretion of Na was increased in dogs injected with venom fraction IV, and is presumed to be due to the inhibition of tubular reabsorption of Na by a natriuretic factor in this venom fraction.

Introduction

Acute renal failure is a common complication of Russell's viper envenomation (Sitprija and Boonpucknavig, 1979; Chugh, 1989). Hemodynamic alterations are among factors incriminated in the pathogenesis of renal failure (Sitprija and Chaiyabutr, 1999). A previous hemodynamic study in dogs following crude Russell's viper venom injection has shown initial hypotension and increased systemic and renal vascular resistance later followed by decreased systemic vascular resistance, increased renal vascular resistance, decreased renal blood flow and glomerular filtration rate (Tungthanathanich et al., 1986; Thamaree et al., 1994). In an isolated renal perfusion study crude Russell's viper venom decreased the glomerular filtration rate in a dose dependent fashion (Ratcliffe et al., 1989). Decreased RBF and GRF due to increased RVR have also been demonstrated in isolated renal perfusion using the venom of Crotalus durissus cascavilla (Martin et al., 1998). Several vasoactive mediators and cytokines from the host have been shown to be involved in hemodynamic changes. Phospholipase A₂ (PLA₂) was believed to be responsible for hypotension through the effect of prostaglandins (Huang and Lee, 1984). Since Russell's viper venom is very complex with several venom components, PLA₂ may not be the only factor that triggers hemodynamic changes. In this report Russell's viper venom was fractionated. The enzymatic activities exhibited by each venom fraction was determined, and the effects of each venom fraction on hemodynamics were studied. The study was descriptive, but served as a basis for further work.

Section snippets

Experimental design

The experiments were performed on 20 healthy anesthetized mongrel male dogs, weighing 10–20 kg and fed on a standard laboratory diet. They were kept in individual cages and given a standard diet and water. Food was withheld 8 h before the experiment. Studies were performed with the approval of and in accordance with recommendations given by the ethics committee of the Thai Red Cross Society.

The dogs were divided into five groups, each group of four dogs. Anesthesia was induced with sodium

Venom fractions

Daboia russellii siamensis venom was separated into five fractions (Fig. 1). Percent protein recovery and enzymatic activities of each venom fraction are displayed in Table 1. Venom fraction III possessed the highest activities of PLA₂, arginine ester hydrolase and proteolytic activity. Venom fraction II had highest phosphomonoesterase, phosphodiesterase and hyaluronidase and second highest proteolytic activities.

Hemodynamics

In each group of dogs maximal changes in hemodynamic parameters including MAP, CO,

Discussion

By Sephadex column chromatography five fractions of Russell's viper venom were isolated. This is a short term study of 2 h of these venom fractions on hemodynamics in dogs. Venom fraction I caused minimal changes in hemodynamic parameters. Striking hemodynamic alterations were produced by venom fractions II, III, IV and V. Profound renal hemodynamic changes were induced by venom fraction III with marked elevation of RVR and a markedly decreased renal blood flow and GFR. These changes were

Acknowledgments

This study was supported by a grant from the Chongkolneenithi Foundation. Miss. Yuppayao Chiemrungsun typed the manuscript.

References (26)

K.S. Chugh
Snake bite induced acute renal failure
Kidney Int.
(1989)
H.C. Huang et al.
Isolation and pharmacological properties of phospholipase A₂ from Vipera russelli (Russell's viper) snake venom
Toxicon
(1984)
O. Khow et al.
Hemorrhagin as a metalloprotease in the venom of Trimeresurus purpureomaculatus: purification and characterization
Toxicon
(2002)
T. Omori-Satoh et al.
Hemorrhagic principles in the venom of Bitis arietans, a viperous snake. I. Purification and characterization
Biochim. Biophys. Acta
(1995)
V.L. Petricevich et al.
Increments in serum cytokine and nitric oxide levels in mice injected with Bothrops asper and Bothrops jararaca snake venoms
Toxicon
(2000)
P. Tungthanathanich et al.
Effect of Russell's viper (Vipera russelli siamensis) venom on renal hemodynamics in dogs
Toxicon
(1986)
K.A. Walsh et al.
Serine protease
Meth. Enzymol.
(1970)
X. Xu et al.
Purification and partial characterization of hyaluronidase from (Agkistrodon acutus) venom
Toxicon
(1982)
C. Buranakarl et al.
Inhibition of renal Na-K-ATPase by Russell's viper venom
Thai. J. Physiol. Sci.
(1997)
N. Chaiyabutr et al.
Effect of starvation on the cardiovascular system, water balance and milk secretion in lactating goats
Res. Vet. Sci.
(1980)

N. Chaiyabutr et al.

Effects of Russell's viper venom on changes of red cell volume in vitro

Thai. J. Vet. Med.

(1987)

A. Chisari et al.

A phospholipase A₂- related snake venom (from Crotalus durissus terrificus) stimulates neuroendocrine and immune functions determination of different sites of action

Endocrinology

(1998)

G.H. De Hass et al.

Purification and properties of phospholipase A from porcine pancreas

Biochim. Biophys. Acta

(1968)

Cited by (17)

The pathophysiological effects of russell's viper (Daboia siamensis) venom and its fractions in the isolated perfused rabbit kidney model: A potential role for platelet activating factor
2020, Toxicon: X
The pathophysiological effects of Russell's viper venom (RVV) and its fractions, including phospholipase A₂ (RvPLA₂), metalloprotease (RvMP), L-amino acid oxidase (RvLAAO), and phosphodiesterase (RvPDE) on renal functions were investigated using the isolated perfused rabbit kidney (IPK) model. Moreover, whether their effects on renal alterations were promoted by platelet activating factor (PAF) was tested using the PAF receptor antagonist, WEB 2086. There was a marked reduction in the perfusion pressure (PP) and renal vascular resistance (RVR) 10 min after RVV administration (1.0 mg/100 ml of perfusate), thereafter both PP and RVR gradually increased and approached the control level within 90 min. These effects were abolished by pretreatment with WEB2086 (2 μg/μl). Administration with RvPLA₂ (280 μg/ml), RvMP (280 μg/ml), or RvLAAO (135 μg/ml) alone increased both the PP and RVR, whereas RvPDE (100 μg/ml) reduced both the PP and RVR. Pretreatment with WEB 2086 completely abolished the effects induced by RvMP, but not the other fractions. The RVV also caused a marked decrease in the glomerular ﬁltration rate (GFR), urinary ﬂow rate (UF), and osmolar clearance (Cosm), and these effects were not inhibited by pretreatment with WEB2086. Each RVV fraction also increased, to varying extents, the GFR, UF, and Cosm, and these effects induced by RvPLA₂ or RvMP, but not the other fractions, were completely blocked by WEB 2086. Changes in percent filtered Na⁺ and K⁺ excreted in the IPK by RVV, RvPDE, and RvMP were abolished by pretreatment with WEB 2086. Histological evaluation proﬁled mainly tubulonephrosis in the treated kidney. These results reveal that the alterations in renal functions induced by RVV and its fractions are due to the synergistic action of the different components of snake venom, instead of the action of a single component. The effects of RVV and its fractions in rabbit IPK are mediated at least in part by PAF.
Determination of the sub-lethal nephrotoxic dose of Russell's viper (Daboia russelii) venom in Wistar rats
2018, Toxicon
Wistar rats were administered increasing doses of Russell's viper venom (RVV; 0.025–0.4 mg/kg) intraperitoneally to investigate acute kidney injury (AKI) by measuring creatinine (1.5-fold increase in serum creatinine above baseline) and examining kidney histology. Approximately 50% of rats receiving 0.25–0.4 mg/kg venom died within 72 h. An increase in serum creatinine only occurred at a venom dose of 0.4 mg/kg, except in two rats. Acute tubular necrosis, glomerular necrosis, cortical necrosis and interstitial inflammation were observed at venom doses of ≥0.25 mg/kg in 12/36 rats. However, of those 12 rats only four survived to 48 h compared to the 24 rats not developing nephrotoxicity, in which 18 were alive at 48 h. There was poor correlation between histological nephrotoxicity and AKI based on creatinine measurement. The early death in rats with AKI makes this a poor model for studying RVV-induced AKI.
Analysis of snake venom metalloproteinases from Myanmar Russell's viper transcriptome
2018, Toxicon
Snake venom metalloproteinases (SVMPs) are the key enzymes in Russell's viper (RV) venom which target all important components of haemostasis, such as clotting factors, platelets, endothelial cells and basement membrane. The structural diversity of SVMPs contributes to the broad spectrum of biological activities. The aim of the study was to investigate the SVMP transcript profile to gain better insights into the characteristic clinical manifestations of the Myanmar Russell's viper (MRV) bites that distinguish it from the RVs of other habitats. Next generation sequencing (RNA-Seq) of mRNA from MRV venom glands (2 males and 1 female) was performed on an Illumina HiSeq2000 platform and then de novo assembled using Trinity software. A total of 59 SVMP contigs were annotated through a Blastn search against the serpent nucleotide database from NCBI. Among them, disintegrins were the most abundant transcripts (75%) followed by the P-III class SVMPs (25%). The P-II SVMPs were scarce (0.002%), while no P-I SVMPs were detectable in the transcriptome. For detailed structural analysis, contigs were conceptually translated and compared with amino acid sequences from other RVs and other vipers using Clustal Omega. The RTS-disintegrin (jerdostatin homolog) was the most abundant among transcripts corresponding to 5 disintegrin isoforms. From 10 isoforms of SVMPs, RVV-X, and Vipera lebetina apoptosis-inducing protease (VLAIP) homolog, hereby termed Daboia siamensis AIP (DSAIP), were found to be highly expressed. Venom protein analysis using SDS-PAGE followed by mass spectrometry revealed that the disintegrin was scarce, while the latter two SVMPs were abundant. These two proteins can contribute to severe clinical manifestations caused by MRV envenomation.
Phosphodiesterase from Daboia russelli russelli venom: Purification, partial characterization and inhibition of platelet aggregation
2014, Toxicon
Citation Excerpt :
Though PDE activity has been reported in the venom of D. russelli siamensis, the role of the enzyme in alteration of haemodynamic system was not properly identified. This is due to heterogeneity of the chromatographic fraction used therein (Suwansrinon et al., 2007). PDE from snake venoms are mostly composed of single polypeptide chain (Dhananjaya and D'Souza, 2010).
Phosphodiesterases (PDEs) belong to a super-family of enzymes that have multiple roles in the metabolism of extracellular nucleotides and regulation of nucleotide-based intercellular signalling. A PDE from Russell's viper (Daboia russelli russelli) venom (DR-PDE) was purified by gel filtration, ion exchange and affinity chromatographies. Homogeneity of the preparation was verified by SDS-PAGE, SE-HPLC and mass spectrometry. It was free from 5′-nucleotidase, alkaline phosphatase and protease activities. Identity of the enzyme was ensured from partial sequence homology with other PDEs. DR-PDE was inactivated by polyvalent anti-venom serum and metal chelators. The enzyme was partially inhibited by the root extracts of four medicinal plants but remained unaffected by inhibitors of intracellular PDEs. DR-PDE hydrolyses ADP and thus, strongly inhibits ADP-induced platelet aggregation in human platelet rich plasma. This study leads to better understanding of a component of Russell's viper venom that affects homoeostatic system of the victim.
Effects of phospholipase A<inf>2</inf> and metalloprotease fractions of Russell's viper venom on cytokines and renal hemodynamics in dogs
2013, Toxicon
Several enzymes in Russell's viper (Daboia siamensis) venom are involved in the venom effects and renal injury. The effects of fractional components of Russell's viper venom, phospholipase A₂ and metalloprotease fractions, were examined in two groups of four experimental dogs each. Animals received an intravenous injection of 140 μg/kg of each venom fraction. The inflammatory effects and renal hemodynamic changes were assessed. Plasma concentrations of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α) and PGE2 were elevated by both phospholipase A₂ (PLA₂) and metalloprotease (MP) fractions. The plasma level of nitric oxide was increased after PLA₂ fraction injection but not with MP fraction injection. Leukocytosis with increase in lymphocytes, monocytes and granulocytes was observed after both PLA₂ and MP injections. Results from this study suggested that both PLA₂ and MP were inflammatory. Increased red blood cell count, hematocrit and hemoglobin concentration were observed in animals injected with PLA₂ fraction, but not with MP fraction. Hemodynamically, PLA₂ fraction induced marked decrease in mean arterial pressure with decreased renal vascular resistance initially followed later by increased renal vascular resistance. MP fraction caused less decrease of mean arterial pressure but increased renal vascular resistance throughout the experiment. Both enzymes decreased renal blood flow, glomerular filtration rate and urine flow. The findings indicate vasodilating effect of PLA₂ fraction and vasoconstricting effect and decreased cardiac function of MP fraction.
Effect of purified Russell's viper venom-factor X activator (RVV-X) on renal hemodynamics, renal functions, and coagulopathy in rats
2011, Toxicon
Citation Excerpt :
It indicates that purified RVV-X may affect the renal hemodynamics via other factors. Changes in renal hemodynamics in rats induced by purified RVV-X were similar to those changes reported in several studies of crude venom injection (Tungthanathanich et al., 1986; Suwansrinon et al., 2007). Decreases in GFR and ERPF and increases in RVR of rats given crude venom and purified RVV-X were possible that the RVV-X as a protease by itself could cause decreased renal blood flow similar to other enzymes such as metalloproteinase.
Acute renal failure (ARF) is the most frequent and a serious complication in victims of Russell’s viper snakebites. Russell’s viper venom-factor X activator (RVV-X) has been identified as a main procoagulant enzyme involving coagulopathy, which might be responsible for changes in renal hemodynamics and renal functions. Here, we purified RVV-X from crude Russell’s viper venom to study renal hemodynamics, renal functions, intravascular clot, and histopathological changes in Sprague–Dawley rats. Changes in renal hemodynamics and renal functions were evaluated by measuring the mean arterial pressure, glomerular filtration rate (GFR), effective renal plasma flow (ERPF), effective renal blood flow (ERBF), renal vascular resistance (RVR), and fractional excretion of electrolytes. After 10 min, rats receiving both crude venom and purified RVV-X decreased GFR, ERPF, and ERBF and increased RVR. These changes correlated to renal lesions. Along with the determination of intravascular clot, rats injected with purified RVV-X increased the average D-dimer level and reached a peak at 10 min, declined temporarily, and then reached another peak at 30 min. The temporal association between clots and renal dysfunction was observed in rats within 10 min after the injection of purified RVV-X. These findings suggested RVV-X as a major cause of renal failure through intravascular clotting in the renal microcirculation.

View all citing articles on Scopus

View full text

Effects of Russell's viper venom fractions on systemic and renal hemodynamics

Abstract

Introduction

Section snippets

Experimental design

Venom fractions

Hemodynamics

Discussion

Acknowledgments

Kidney Int.

Toxicon

Toxicon

Biochim. Biophys. Acta

Toxicon

Toxicon

Meth. Enzymol.

Toxicon

Inhibition of renal Na-K-ATPase by Russell's viper venom

Thai. J. Physiol. Sci.

Effect of starvation on the cardiovascular system, water balance and milk secretion in lactating goats

Res. Vet. Sci.

Effects of Russell's viper venom on changes of red cell volume in vitro

Thai. J. Vet. Med.

A phospholipase A2- related snake venom (from Crotalus durissus terrificus) stimulates neuroendocrine and immune functions determination of different sites of action

Endocrinology

Purification and properties of phospholipase A from porcine pancreas

Biochim. Biophys. Acta

A phospholipase A₂- related snake venom (from Crotalus durissus terrificus) stimulates neuroendocrine and immune functions determination of different sites of action