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Journal of Biological Sciences

Year: 2016 | Volume: 16 | Issue: 4 | Page No.: 112-119
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ABSTRACT

Background: Scientists are exploring the application of new drugs derived from plant sources as an alternative to snakebite anti-venom therapy, which is endowed with several limitations. Naturally produced antioxidants like the flavonoids, which are widely found in the plants are largely consumed in daily diet, at present. Methodology: This study investigates and evaluates the potential effects of some flavonoids (quercetin) on envenoming of albino rats by sub-lethal venom (3.84 mg kg–1, i.p.) doses of Echis coloratus (Ec) viper snake. Beneficial effects of quercetin (30 μM kg–1, i.p.) doses on the induced hepato-renal toxicity are assessed by the measurement of selected biomarkers. Results: Sacrificed animals sera and tissues, which were collected for biochemical studies, showed significant increase in the levels of AST, ALT, ALP and creatinine. There was significant histopathological damage of the tissue's architecture and rise in the liver and kidney MDA levels.Conclusion: It is concluded that the reversal effects of modulation of the biochemical parameters and histological damage are attributed to the potential protective effects that ensued after administration of quercetin.
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Received: February 14, 2016;   Accepted: March 06, 2016;   Published: April 15, 2016

INTRODUCTION

Hot weather places of the world are endowed with many snakes that belong to the carnivorous, poikilothermic reptiles. Vipers, like E. coloratus have vasculotoxic venom component that deteriorate the human vascular system. Those ones found in the central region of Saudi Arabia have not been studied thoroughly, especially their venoms that have received little attention1. Several factors have different physiological and biological effects on the bitten victims. Components of viper venoms have mainly enzymatic activities that break intracellular organelles, leading to necrosis, edema and hemorrhage2 that culminate into loss of several body organs due to dysfunction tissue loss3-5.

It was shown that several viper venoms constituents that are characterized by high molecular weights will show a slow absorption pattern through the lymphatic system6,7. Victims that have been envenomed usually show symptoms that vary according to type, size and sex of the snake and bitten victim, in addition to environmental, venom and treatment factors8,9.

Several investigators have reflected the after snake envenoming signs and symptoms as several clinical abnormalities. The development of acute hepato-renal toxicity10, organ dystrophy11, cytotoxic cellular manifestations12 and hypoglycemia13 metabolic complications14 all are induced acutely by envenoming bites.

Some studies have suggested that treatment by specific antivenins decrease the extent of the hemostatic failure, but still the management of E. coloratus victims remains uncertain. Victims subjected to such venoms and treated non-specifically reflected extremely urgent problems associated with antitoxic substances. This situation only stressed the queries and barriers associated with antivenin therapy that put researchers and scientists on the alert of finding alternative new drugs of plant and herbal origin to treat snakebiteand replace or limit the untoward effects brought about by antivenin use15.

Recently, envenoming herbal treatment received more concentrated attention by subjecting traditionally used medical plants to pharmacological screening and isolating their active components16. Herbal extracts like quercetin has anti-inflammatory, polyphenolic antioxidant17, angio protective activity18,19 and appreciated beneficial effects on the immune system20. Further, extra benefits are associated with cell membrane stabilization21, calcium ions transport22 and membrane alteration23.

In this present study, aim to investigate the biochemical activity mechanisms of quercetin and its beneficial neutralizing and ameliorating hepato-renal toxicity effects induced by E. coloratus viper venom in rats.

MATERIALS AND METHODS

Materials: Adult male albino Wistar rats (200-220 g), E. coloratus lyophilized snake venom, quercetin-3,3’,4’,5,6 pentahydroxyflavone’ 2-(3,4-Dihydroxyphenyl)-3,5,7-trihydroxy-4H-1-benzopyran-4-one (Sigma, USA) and other laboratory chematerials.

Preparation of venom solution: Echis coloratus lyophilized snake venom were obtained from Research Department of the Prince Sultan Riyadh Military Medical City, Riyadh, Saudi Arabia. The venom was dissolved in physiological saline (0.9% NaCl) (final concentration 10 mg mL–1) under mild mixing for 10 min at 4°C and then, centrifuged at 10,000 rpm for 10 min at 4°C. The pellet was discarded and the supernatant was aliquoted and stored at -20°C at a maximum time of one week and administered intraperitoneally (i.p.) at a dose of 3.84 mg kg–1 b.wt. (LD50) in a maximum volume of 0.2 mL (Fig. 1).

Preparation of quercetin: Quercetin -3,3’,4’,5,6 Pentahydroxyflavone’ 2-(3,4-Dihydroxyphenyl)-3,5,7-trihydroxy-4H-1-benzopyran-4-one (Sigma, USA) was dissolved in 1 M NaOH and administration by intraperitoneally (i.p.) at a dose of 30 μM kg–1 b.wt., in a maximum volume of 0.2 mL (Fig. 2).

Image for - Beneficial Role of Quercetin on Echis coloratus Snake Venom Induced Hepato-renal Toxicity in Rats
Fig. 1:Echis coloratus snake venom

Image for - Beneficial Role of Quercetin on Echis coloratus Snake Venom Induced Hepato-renal Toxicity in Rats
Fig. 2: Structure of quercetin

Animals: Adult male albino Wistar rats with weight between 200-220 g were used for this study. Animals were kept under standardized conditions of temperature (22±1°C), humidity (55±5%) and 12-12 h light-dark cycles with free access to food and water.

Methods: The animals were divided into four groups of six rats. Control group I were injected intraperitoneally (i.p.) with 200 μL normal saline (0.9% NaCl) only. Group II were injected (i.p.) with 200 μL crude venom (3.84 mg kg–1) alone. Group III were injected (i.p.) with 200 μL venom (3.84 mg kg–1) and quercetin (30 μM kg–1). Group IV were injected (i.p.) with 200 μL quercetin alone.

One hour later, injected animals were decapitated and blood sample were collected for biochemical analysis and tissue organs were collected for histopathological analysis.

Ethics statement: Experiments were done according to the Saudi Arabian law of protection of animals. The study procedures involving animal experiments were approved by the institutional ethics committee.

Biochemical analysis: Blood sample were collected from each rat into plain centrifuge tubes, left for 1 h at room temperature (25°C±2) and serum were separated by centrifugation at 600×g for 15 min and analyzed, without delay for the concentration of creatinine, alanine aminotransferase (ALT), aspartate aminotransferase (AST) and alkaline phosphatase (ALP) were determined by using kits purchased from United Diagnostic Industry (UDI) Dammam, Saudi Arabia. A few pieces of liver and kidney were fixed with 10% neutral buffered formalin for histopathological investigations, whereas the majority of pieces were homogenized in ice-cold medium containing 50 mM tris-HCl and 300 mM sucrose, pH 7.4 and finally stored at -80°C until use in the various biochemical determinations.

Determination of TBARS (oxidative stress): The levels of TBARS, a marker of lipid peroxidation24-26 were estimated in liver and kidney tissues using the method of Niehaus and Samuelsson27. Tissues were homogenized in tris-HCl buffer (pH 7.5) followed by centrifugation to remove the cellular debris. An aliquot (100 μL) of supernatant from the tissue homogenate was mixed with 2 mL of TBA-TCA-HCl (1:1:1) reagent (0.37% TBA, 15% TCA and 0.25 N HCl) and the tubes were placed in a water bath at 90°C for 10 min. The tubes were allowed to cool at room temperature and then the absorbance of the colored solution was measured against a reagent blank at 535 nm.

Histopathological studies: Tissue samples were fixed in 10% neutral formalin for 24 h and paraffin blocks were prepared and routinely processed for light microscopy. Slices of 4-5 μm were obtained from the prepared blocks and stained with hematoxylin-eosin. The preparations obtained were visualized using a Nikon microscopy at a magnification of 400×.

Statistical analysis: Results are presented as Mean±SEM. Comparisons were done by two-tail unpaired Student’s t-test or one-way ANOVA as appropriate. The p≤0.05 were considered significant.

RESULTS

Hepato-renal function biomarkers: The administration of Echis coloratus venom to the rats caused significant increase in AST, ALT and ALP and blood serum creatinine levels, when it was compared with control group. Co-administration of quercetin decreased the rise in AST, ALT and ALP and blood serum creatinine levels in Fig. 3-6, respectively.

Image for - Beneficial Role of Quercetin on Echis coloratus Snake Venom Induced Hepato-renal Toxicity in Rats
Fig. 3:
Effect of the treatments of the groups in serum AST activity, values is expressed as Mean±SEM (n = 6). Results are represented in the form of Mean±SE, *Significant at p<0.05

Image for - Beneficial Role of Quercetin on Echis coloratus Snake Venom Induced Hepato-renal Toxicity in Rats
Fig. 4:
Effect of the treatments of the groups in serum ALT activity, values is expressed as Mean±SEM (n = 6). Results are represented in the form of Mean±SE, *Significant at p<0.05

Image for - Beneficial Role of Quercetin on Echis coloratus Snake Venom Induced Hepato-renal Toxicity in Rats
Fig. 5:
Effect of the treatments of the groups in serum ALP activity, values is expressed as Mean±SEM (n = 6). Results are represented in the form of Mean±SE, *Significant at p<0.05

Image for - Beneficial Role of Quercetin on Echis coloratus Snake Venom Induced Hepato-renal Toxicity in Rats
Fig. 6:
Effect of the treatments of the groups in serum creatinine activity, values is expressed as Mean±SEM (n = 6). Results are represented in the form of Mean±S.E, *Significant at p<0.05

Oxidative stress: In the liver and kidney tissues, MDA was significantly increased in Echis coloratus venom treated group as compared to control group and this rise in MDA was decreased by quercetin (Table 1).

Histology: The histology of the liver tissue of the control and quercetin-treated animals showed normal histological structure of hepatocytes, central vein blood sinusoid and nuclei (Fig. 7), whereas in venom group showed cellular infiltrations, degenerative changes of hepatic cells with cell necrosis and disarrangement of normal hepatic cells were observed. The histology of liver tissues of venom+quercetin group showed less degeneration, necrosis and disarrangement of normal hepatic cells (Fig. 7).

The histology of the kidney tissue of control and quercetin-treated animals showed normal morphological appearances (Fig. 8), whereas the venom group showed congestion, necrosis and degeneration in the epithelial cells of renal tubules and swelling in the lining endothelium of the glomerulus.

Table 1:Effect of quercetin on Echis coloratus venom induced MDA levels in rat tissue
Image for - Beneficial Role of Quercetin on Echis coloratus Snake Venom Induced Hepato-renal Toxicity in Rats
Results are represented in the form of Mean±SE, *Significant at p<0.05, Values expressed as Nm of MDA g–1 weight of tissue

The histology of kidney tissues of venom+quercetin group showed less congestion, necrosis, degeneration in the epithelial cells of renal tubules and swelling in the lining endothelium of the glomerulus (Fig. 8).

DISCUSSION

Human hemostasis is severely deteriorated by E. coloratus envenoming and it was reported that anemia ensued due to excessive bleeding in about a third of the victims28. In this study, it was noticed that envenomed rats showed hemorrhagic signs and spots in different organs after sacrifice, similar to the reported observations by other investigators9. Such observations could be attributed to factors are coagulopathic and defibrinating in content9. Organs with hemorrhagic areas are formed due to their cells basal membrane alterations of the endothelial lining vascular permeability that allowed blood to ooze into tissues in the vicinity. Envenoming by E. coloratus usually allows for the formation of thrombotic hepatic portal vein leading to blood vessels congestion and damage of the cell membrane endothelial lining. On the other hand, phospholipase A2 enzyme releases plasma lysolecithin and also directly destroys cell membranes9. Venoms of such vipers like E. coloratus and limited numbers of other snakes have the ability to form thrombin by precursors of prothrombin29.

Liver function markers (ALT, AST and ALP) reflected hepatic dysfunction by showing significant increase of their activity levels in rat sera, in comparison with control groups. This situation is speculated and envisaged through free radicals production induced by E. coloratus viper envenoming that led to hepatotoxic oxidative stress and the ensuing extra hepatic obstruction and damage in liver function30,31. Rise in liver enzymes that followed viper venom administration is indicative of hepatocellular injury32. Hepatocytes mitochondria get congested with liver enzymes and the biochemical indices increased due to the altered transportation input/output of the mitochondrial membrane33-36.

On the other hand the induced renal stress was viewed through elevation of the serum creatinine, which is a marker of nephritis37-42. As a protein metabolism waste product that is normally excreted by kidneys, creatinine excess levels in serum affirms some degree of deficient kidney function43 as a specific kidney marker44.

Image for - Beneficial Role of Quercetin on Echis coloratus Snake Venom Induced Hepato-renal Toxicity in Rats
Fig. 7(a-d):
Effect of quercetin on snake venom induced histological changes of liver tissue. Quercetin enhanced the protective mechanism of the liver and helped in the restoration of the damaged histology. Representative sections of the liver of (a) Normal rats, (b) Venom treated rats, (C) Quercetin treated rats and (d) Pretreated with quercetin+venom

Image for - Beneficial Role of Quercetin on Echis coloratus Snake Venom Induced Hepato-renal Toxicity in Rats
Fig. 8(a-d):
Effect of quercetin on snake venom induced histological changes of liver tissue. Quercetin enhanced the protective mechanism of the kidney and helped in the restoration of the damaged histology. Representative sections of the kidney of (a) Normal rats, (b) Venom treated rats, (c) Quercetin treated rats and (d) Pretreated with quercetin+venom

The out coming results of the excess hepatic and nephrotic tissue MDA levels also confirm the oxidative stress and lipid peroxidation indication45 and free radicals action46. Extensive explanation of the free radicals production and various mechanisms of actions and the countering antioxidant defense mechanisms were previously reported47,48. This has been modulated in several recent studies49-55.

The scavenging ability of quercetin56 and polyphenol compounds57 convey protective effects against the actions of venoms and free radicals. Confirming similar studies on rats using Vipera russelli venom were previously done by Ali et al.58.

The findings of the present study confirm the outcome of previous studies on mice MDA levels53 mechanisms involved in lipid peroxidation, fatty acids, adipose tissue59,60 and inhibition of PLA261 and membrane stabilization improvement62,63. Quercetin could have restored the balance between the venom-induced free radicals and the defensive antioxidant system mechanisms with the tissue redox state64. The massive cellular activity and mechanisms involved in the associated venom detoxification and nuclear interruption lead to appreciable karyorrhexis and pyknosis in the nuclei, in the outcome of this present study that reflected the cellular changes and necrosis, following administration of E. coloratus venom. Such findings were also concluded following the employment of other venoms (cobra snakes) done by Rahmy et al.65.

Kidneys histological tests confirmed the renal stress evaluated by sera analysis, viewed in the tissue alterations. Further, cortical renal necrosis and related to intravascular coagulation added to vasospasm toxic effect action mechanisms were previously discussed9,66.

CONCLUSION

The present experimental results indicate that quercetin was beneficial in countering the toxic effects of E. coloratus venom and or has an alternative or complementary treatment strategy of envenoming. Further experimental approaches could address quercetin, which could possibly lead to the development of pharmaceutical formulations for treating snake bite victims.

ACKNOWLEDGMENTS

The authors express their appreciation and thanks to the administration of Prince Sultan Military Medical City for their encouragement and support. The support and help of our colleagues Dr. Abdulquaiyoom Khan, Mr. Dhaya Sankar, Mr. Tariq Al Zamil and Ms. Ruba Fahad Al Mohini is thankfully acknowledged.

REFERENCES

  1. Al-Sadoon, M.K., 1989. Survey of the reptilian fauna of Saudi Arabia. 1-The snake fauna of the central region. J. King Saud Univ. Sci., 1: 53-69.
  2. Gutierrez, J.M., 1995. Clinical Toxicology of Snake Bite in Central America. In: Handbook of Clinical Toxicology of Animal Venoms and Poisons, White, J. and J. Meier (Eds.). CRC Press, Boca Raton FL., ISBN: 9780849344893, pp: 645-665
  3. Abdel-Nabi, I., R. Awadalla and I. El-Shamy, 1997. Biochemical effects of intraperitoneal injection of rats with the venom of the snake Echis carinatus. Egypt. J. Zool., 29: 195-205.
  4. Marsh, N., D. Gattullo, P. Pagliaro and G. Losano, 1997. The Gaboon viper, Bitis gabonica: Hemorrhagic, metabolic, cardiovascular and clinical effects of the venom. Life Sci., 61: 763-769.
    CrossRefPubMedDirect Link
  5. Masood, M.F., 2012. Ecological studies on the diversity of terrestrial poisonous snakes proteroglyphous of Jazan region Kingdom of Saudi Arabia (Reptilia: Ophidia). Egypt. J. Hosp. Med., 49: 839-856.
    Direct Link
  6. Reid, H.A. and R.D.G. Theakston, 1983. The management of snake bite. Bull. World Health Organ., 61: 885-895.
    Direct Link
  7. Warrell, D.A., 1995. Clinical Toxicology of Snake Bites in Africa and the Middle East/Arabian Peninsula. In: Handbook of Clinical Toxicology of Animal Venoms and Poisons, White, J. and J. Meier (Eds.). CRC Press, Boca Raton FL., ISBN: 9780849344893, pp: 433-492
  8. Latifi, M., 1984. The Snakes of Iran. Iran Department of the Environment, Tehran, pp: 221.
  9. Jarrar, B.M., 2011. Histological alterations and biochemical changes in the liver of sheep following Echis coloratus envenomation. Saudi J. Biol. Sci., 18: 169-174.
    CrossRefDirect Link
  10. Ickowicz, M., A. Shulov and D. Naor, 1966. The effect of Vipera palestina venom on the thymus, lymph nodes and kidneys. Toxicon, 3: 305-306.
    CrossRefPubMedDirect Link
  11. Mohamed, A.H. and L.Z. Khaled, 1966. Effect of the venom of Cerastes cerastes on nerve tissue and skeletal muscle. Toxicon, 3: 223-224.
    CrossRefPubMedDirect Link
  12. Bertke, E.M. and J.H. Atkins, 1961. Effect of Centruroides sculpturatus venom upon rat tissue: A histopathologic study. Toxicon, 2: 205-209.
    CrossRefDirect Link
  13. Abu-Sinna, G., A.S. Al-Zahaby, A.A. El-Aal, A.A. El-Baset and N.A. Soliman, 1993. The effect of the viper Cerastes cerastes cerastes venom and venom fractions on carbohydrate metabolism. Toxicon, 31: 791-801.
    CrossRefDirect Link
  14. Mahmoud, I., 1983. Biochemical and physiological studies on the action of viper venom Cerastes vipera on mice Mus musculus. Masters Thesis, Fac. Sci. Cairo University, Egypt.
  15. Premendran, S.J., K.J. Salwe, S. Pathak, R. Brahmane and K. Manimekalai, 2011. Anti-cobra venom activity of plant Andrographis paniculata and its comparison with polyvalent anti-snake venom. J. Nat. Sci. Biol. Med., 2: 198-204.
    CrossRefDirect Link
  16. Mukherjee, A.K., 2012. Green medicine as a harmonizing tool to antivenom therapy for the clinical management of snakebite: The road ahead. Indian J. Med. Res., 136: 10-12.
    Direct Link
  17. McAnlis, G.T., J. McEneny, J. Pearce and I.S. Young, 1999. Absorption and antioxidant effects of quercetin from onions, in man. Eur. J. Clin. Nutr., 53: 92-96.
    PubMedDirect Link
  18. Atalik, K.E., B. Keles, Y. Uyar, M.A. Dundar, M. Oz and H.H. Esen, 2010. Vasoprotection by melatonin and quercetin in rats treated with cisplatin. Indian J. Exp. Biol., 48: 1188-1193.
    Direct Link
  19. Jung, J.H., J.I. Kang and H.S. Kim, 2012. Effect of quercetin on impaired immune function in mice exposed to irradiation. Nutr. Res. Pract., 6: 301-307.
    CrossRefDirect Link
  20. Lee, S., H.S. Park, Y. Notsu, H.S. Ban and Y.P. Kim et al., 2008. Effects of hyperin, isoquercitrin and quercetin on lipopolysaccharide-induced nitrite production in rat peritoneal macrophages. Phytother. Res., 22: 1552-1556.
    CrossRefPubMedDirect Link
  21. Margina, D., M. Ilie, G. Manda, I.Neagoe and M. Mocanu et al., 2012. Quercetin and epigallocatechin gallate effects on the cell membranes biophysical properties correlate with their antioxidant potential. Gen. Physiol. Biophys., 31: 47-55.
    PubMedDirect Link
  22. Nishino, H., E. Naitoh, A. Iwashima and K. Umezawa, 1984. Quercetin interacts with calmodulin, a calcium regulatory protein. Experientia, 40: 184-185.
    CrossRefDirect Link
  23. Margina, D., M. Ilie, C. Negrei, D. Gradinaru, A. Balanescu and N. Mitrea, 2010. Quercetin and epigallocatechin gallate in vitro induced changes in the membrane anisotropy of peripheral blood mononuclear cells from patients with inflammatory diseases. J. Med. Plants Res., 4: 2388-2392.
    Direct Link
  24. Khan, H.A., M.A.K. Abdelhalim, M.S. Al-Ayed and A.S. Alhomida, 2012. Effect of gold nanoparticles on glutathione and malondialdehyde levels in liver, lung and heart of rats. Saudi J. Biol. Sci., 19: 461-464.
    CrossRefDirect Link
  25. Al Deeb, S., K. Al Moutaery, H.A. Khan and M. Tariq, 2000. Exacerbation of iminodipropionitrile-induced behavioral toxicity, oxidative stress and vestibular hair cell degeneration by gentamicin in rats. Neurotoxicol. Teratol., 22: 213-220.
    CrossRefDirect Link
  26. Tariq, M., H.A. Khan, Z. Rehana, K. Al Moutaery and S. Al Deeb, 1998. Proglumide, a cholecystokinin receptor antagonist, exacerbates β, β'-iminodipropionitrile-induced dyskinetic syndrome in rats. Neurotoxicol. Teratol., 20: 571-579.
    CrossRefDirect Link
  27. Niehaus, Jr. W.G. and B. Samuelsson, 1968. Formation of malonaldehyde from phospholipid arachidonate during microsomal lipid peroxidation. Eur. J. Biochem., 6: 126-130.
    CrossRefPubMedDirect Link
  28. Benbassat, J. and O. Shalev, 1993. Envenomation by Echis coloratus (Mid-East saw-scaled viper): A review of the literature and indications for treatment. Israel J. Med. Sci., 29: 239-250.
    PubMedDirect Link
  29. Chang, C.C., 1979. The Action of Snake Venoms on Nerve and Muscle. In: Snake Venoms (Handbook of Experimental Pharmacology, Volume 52), Lee, C.Y. (Ed.). Chapter 10, Springer-Verlag, USA., ISBN: 978-3-642-66915-6, pp: 310-312
  30. Hatoff, D.E. and W.J. Hardison, 1980. Hepatic bile acid content control alkaline phosphatase during cholestasis. Gastroenterology, 78: 1307-1307.
  31. Sherlock, H., 1981. Sherlock's Diseases of the Liver and Biliary System. 6th Edn., Blackwell Scientific Publications, Oxford, UK., ISBN: 9780632007660, pp: 4, 42-44.
  32. Kalender, S., A. Ogutcu, M. Uzunhisarcikli, F. Acikgoz, D. Durak, Y. Ulusoy and Y. Kalender, 2005. Diazinon-induced hepatotoxicity and protective effect of vitamin E on some biochemical indices and ultrastructural changes. Toxicology, 211: 197-206.
    CrossRefDirect Link
  33. Teimouri, F., N. Amirkabirian, H. Esmaily, A. Mohammadirad, A. Aliahmadi and M. Abdollahi, 2006. Alteration of hepatic cells glucose metabolism as a non-cholinergic detoxication mechanism in counteracting diazinon-induced oxidative stress. Hum. Exp. Toxicol., 25: 697-703.
    CrossRefDirect Link
  34. Kappers, W.A., R.J. Edwards, S. Murray and A.R. Boobis, 2001. Diazinon is activated by CYP2C19 in human liver. Toxicol. Applied Pharmacol., 177: 68-76.
    CrossRefDirect Link
  35. Sams, C., J. Cocker and M.S. Lennard, 2003. Metabolism of chlorpyrifos and diazinon by human liver microsomes. Toxicol. Lett., 144: s146-s146.
    CrossRefDirect Link
  36. Ogutcu, A., M. Uzunhisarcikli, S. Kalender, D. Durak, F. Bayrakdar and Y. Kalender, 2006. The effects of organophosphate insecticide diazinon on malondialdehyde levels and myocardial cells in rat heart tissue and protective role of vitamin E. Pestic. Biochem. Physiol., 86: 93-98.
    CrossRefDirect Link
  37. Yousef, M.I., F.M. El-Demerdash, K.I. Kamel and K.S. Al-Salhen, 2003. Changes in some hematological and biochemical indices of rabbits induced by isoflavones and cypermethrin. Toxicology, 189: 223-234.
    CrossRefPubMedDirect Link
  38. Eraslan, G., A. Bilgili, D. Essiz, M. Akdogan and F. Sahindokuyucu, 2007. The effects of deltamethrin on some serum biochemical parameters in mice. Pestic. Biochem. Physiol., 87: 123-130.
    CrossRefDirect Link
  39. Omurtag, G.Z., A. Tozan, A.O. Sehirli and G. Sener, 2008. Melatonin protects against endosulfan-induced oxidative tissue damage in rats. J. Pineal. Res., 44: 432-438.
    CrossRefPubMed
  40. El-Shenawy, N.S., R.A. Al-Eisa, F. El-Salmy and O. Salah, 2009. Prophylactic effect of vitamin E against hepatotoxicity, nephrotoxicity, haematological indices and histopathology induced by diazinon insecticide in mice. Curr. Zool., 55: 219-226.
    Direct Link
  41. Al-Attar, A.M. and W.A. Al-Taisan, 2010. Preventive effects of black seed (Nigella sativa) extract on sprague Dawley rats exposed to diazinon. Aust. J. Basic Applied Sci., 4: 957-968.
    Direct Link
  42. Sarhan, O.M.M. and Z.Y. Al-Sahhaf, 2011. Histological and biochemical effects of diazinon on liver and kidney of rabbits. Life Sci. J., 8: 1183-1189.
    Direct Link
  43. Panda, N.C., 1999. Kidney. In: Textbook of Biochemistry and Human Biology, Talwar, G.P. and L.M. Srivastava (Eds.). 2nd Edn., Prentice Hall, India, pp: 290-296
  44. Cheesbrough, M., 1998. Clinical Chemistry Tests. In: District Laboratory Practice in Tropical Countries: Part 1, Cheesbrough, M. (Ed.). Cambridge University Press, Cambridge, pp: 331-363
  45. Jacob, R.A. and B.J. Burri, 1996. Oxidative damage and defense. Am. J. Clin. Nutr., 63: 985S-990S.
    PubMedDirect Link
  46. Esterbauer, H., R.J. Schaur and H. Zollner, 1991. Chemistry and biochemistry of 4-hydroxynonenal, malonaldehyde and related aldehydes. Free Radic. Biol. Med., 11: 81-128.
    CrossRefPubMedDirect Link
  47. Halliwell, B., 1991. Drug antioxidant effects. A basis for drug selection. Drugs, 42: 569-605.
    CrossRefPubMedDirect Link
  48. Ray, S., 2012. Evaluation of protective role of morin on busulfan-induced lipid peroxidation. Int. J. PharmTech Res., 3: 2222-2227.
    Direct Link
  49. Altuntas, I. and N. Delibas, 2002. The effects of fenthion on lipid peroxidation and some liver enzymes: The possible protective role of vitamins E and C. Turk. J. Med. Sci., 32: 293-297.
    Direct Link
  50. Mansour, S.A and A.H. Mossa, 2005. Comparative effects of some insecticides as technical and formulated on male albino rats. J. Egypt. Soc. Toxicol., 32: 41-54.
  51. Fortunato, J.J., F.R. Agostinho, G.Z. Reus, F.C. Petronilho, F. Dal-Pizzol and J. Quevedo, 2006. Lipid peroxidative damage on malathion exposure in rats. Neurotoxicity Res., 9: 23-28.
    CrossRefDirect Link
  52. Akturk, O., H. Demirin, R. Sutcu, N. Yilmaz, H. Koylu and I. Altuntas, 2006. The effects of diazinon on lipid peroxidation and antioxidant enzymes in rat heart and ameliorating role of vitamin E and vitamin C. Cell Biol. Toxicol., 22: 455-461.
    CrossRefPubMedDirect Link
  53. Al Asmari, A., K. Al Moutaery, R.A. Manthari and H.A. Khan, 2006. Time-course of lipid peroxidation in different organs of mice treated with Echis pyramidum snake venom. J. Biochem. Mol. Toxicol., 20: 93-95.
    CrossRefDirect Link
  54. Amirkabirian, N., F. Teimouri, H. Esmaily, A. Mohammadirad, A. Aliahmadi and M. Abdollahi, 2007. Protection by pentoxifylline of diazinon-induced toxic stress in rat liver and muscle. Toxicol. Mech. Methods, 17: 215-221.
    CrossRefDirect Link
  55. Shah, M.D. and M. Iqbal, 2010. Diazinon-induced oxidative stress and renal dysfunction in rats. Food Chem. Toxicol., 48: 3345-3353.
    CrossRefPubMedDirect Link
  56. Bors, W., C. Michel and M. Saran, 1994. Flavonoid antioxidants: Rate constants for reactions with oxygen radicals. Methods Enzymol., 234: 420-429.
    CrossRefPubMedDirect Link
  57. Kitagawa, S., H. Sakamoto and H. Tano, 2004. Inhibitory effects of flavonoids on free radical-induced hemolysis and their oxidative effects on hemoglobin. Chem. Pharmaceut. Bull., 52: 999-1001.
    CrossRefPubMedDirect Link
  58. Ali, S.F., M. Tariq, M. Hasan and S.S. Haider, 1981. Effect of Russell's venom on lipid peroxidation in organs of the mouse. Toxicon, 19: 903-905.
    CrossRefPubMedDirect Link
  59. El-Asmar, M.F., R.M. Farag, S. Shoukry and I.R. El-Smimi, 1979. Effect of scorpion (Leiurus quinquestriatus H and E) venom on lipid metabolism. Toxicon, 17: 279-283.
    CrossRefPubMedDirect Link
  60. Yarom, R. and K. Braun, 1970. Cardiovascular effects of scorpion venom, morphological changes in the myocardium. Toxicon, 8: 41-46.
    CrossRefPubMedDirect Link
  61. Borowitz, S.M. and C. Montgomery, 1989. The role of phospholipase A2 in microsomal lipid peroxidation induced with t-butyl hydroperoxide. Biochem. Biophys. Res. Commun., 158: 1021-1028.
    CrossRefPubMedDirect Link
  62. Mukherjee, A.K. and C.R. Maity, 1998. Effect of dietary supplementation of vitamin E in partial inhibition of Russell's viper venom phospholipase A2 induced hepatocellular and microsomal membrane damage in rats. Acta Physiologica Hungarica, 85: 367-374.
    PubMedDirect Link
  63. Alam, M.I. and A. Gomes, 1998. Viper venom-induced inflammation and inhibition of free radical formation by pure compound (2-hydroxy-4-methoxy benzoic acid) isolated and purified from anantamul (Hemidesmus indicus R. BR) root extract. Toxicon, 36: 207-215.
    CrossRefPubMedDirect Link
  64. Schmitt-Schillig, S., S. Schaffer, C.C. Weber, G.P. Eckert and G.P. Muller, 2005. Flavonoids and the aging brain. J. Physiol. Pharmacol., 56: 23-36.
    PubMedDirect Link
  65. Rahmy, T.R., M.H. El-Naggar and A.E. Mehana, 2005. Immunohistochemical detection of hepatic injury after cobra snake envenoming. Egypt. J. Nat. Toxins, 2: 119-134.
    Direct Link
  66. Al-Sadoon, M.K. and A. Fahim, 2012. Possible recovery from an acute envenomation in male rats with LD50 of Echis coloratus crude venom: I-A seven days hematological follow-up study. Saudi J. Biol. Sci., 19: 221-227.
    CrossRefPubMedDirect Link

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