Archives of Medicine and Health Sciences

ORIGINAL ARTICLE
Year
: 2017  |  Volume : 5  |  Issue : 2  |  Page : 200--203

Effect of vitamin E supplementation on superoxide and malondialdehyde generation in acute celphos poisoning


C Biwas1, J Bala2, Simmi Kharb2,  
1 Department of Biochemistry, Government Medical College, Agartala, Tripura, India
2 Department of Biochemistry, Pt. B.D. Sharma University of Health Sciences, Rohtak, Haryana, India

Correspondence Address:
Simmi Kharb
H.No. 1396, Sector-1, Rohtak, Haryana
India

Abstract

Introduction: Aluminum phosphide is one of the most commonly used grain fumigants and aluminum phosphide poisoning (ALP) has been reported as the most common cause of acute poisoning in India. Aluminum toxicity has been reported to increase the rate of lipid peroxidation and free radical formation. Materials and Methods: The present study was designed to investigate the role of vitamin E supplementation on free radical generation and lipid peroxidation in acute aluminium phosphide poisoning in rats. Thirty disease free albino rats were taken to study the effect of acute aluminium phosphide poisoning (ALP poisoning) were further divided into 3 subgroups of ten rats each: A, B and C. Group A: given vehicle (Ginni Oil) only. Group B: given 5 ml 'celphos mixture' (or 0.3mg/g body wt.). Group C: rats with acute Celphos poisoning along with vitamin E (1.5 mg vitamin E/g body weight of rat. The MDA levels and superoxide levels (Nitroblue tetrazolium (NBT) reduction) were estimated. Results: MDA levels were significantly higher in the group B as compared to Group A. In group C, administration of vitamin E resulted in decreased MDA level compared to group B. MDA levels in group C still remained significantly higher as compared to group A. NBT reduction was significantly increased in group B as compared to group A. Administration of vitamin E to rats of group C resulted in significant decrease of NBT reduction. Conclusion: Findings of the present study showed that vitamin E via its antioxidant action and anti-inflammatory effects has protective effect on phosphine-induced toxicity in rats.



How to cite this article:
Biwas C, Bala J, Kharb S. Effect of vitamin E supplementation on superoxide and malondialdehyde generation in acute celphos poisoning.Arch Med Health Sci 2017;5:200-203


How to cite this URL:
Biwas C, Bala J, Kharb S. Effect of vitamin E supplementation on superoxide and malondialdehyde generation in acute celphos poisoning. Arch Med Health Sci [serial online] 2017 [cited 2020 Mar 28 ];5:200-203
Available from: http://www.amhsjournal.org/text.asp?2017/5/2/200/220825


Full Text



 Introduction



Aluminum phosphide (ALP) is one of the most commonly used grain fumigants because of its advantageous properties such as it is toxic to all stages of insects, is highly potent, does not affect seed viability, is free from toxic residues, and leaves little residue on food grains. ALP poisoning has been reported as the most common cause of acute poisoning in India.[1] Accidental and suicidal death are common in rural Northwest and Central India, which is mainly attributable to poor regulation regarding the accessibility of this gravely toxic rodenticide.[2]

ALP after coming in contact with moisture forms phosphine (PH3) gas which leads to poisoning on inhalation, ingestion, and dermal contact.[3] After oral intake, the PH3 gas released and absorbed by the gastrointestinal tract with simple diffusion and is mainly excreted by the kidneys and lungs. PH3, like cyanide, inhibits mitochondrial cytochrome oxidase and cellular oxygen utilization.[4],[5],[6] PH3 is excreted through the breath and urine as hypophosphite and inhibits the electron transport resulting from noncompetitive inhibition of enzyme cytochrome oxidase of the mitochondria, leading to respiratory chain inhibition which leads to cellular hypoxia and small vessel injury, which is further potentiated by cardiotoxicity due to anoxic myocardial damage and shock.[7] PH3-mediated myocardial contractility and fluid loss result in pulmonary edema. Hence, metabolic acidosis, respiratory alkalosis, acute renal failure, disseminated intravascular coagulation, hepatic necrosis, and hypo- or hyper-magnesemia may also occur.[8] Kidney is one of the major targets of PH3 poisoning in the human body. The increase in blood urea had been creatinine is reported in previous studies due to ALP.[9]

Aluminum toxicity has been reported to increase the rate of lipid peroxidation and free radical formation. Researchers have shown that ingestion of ALP leads to high superoxide dismutase (SOD) activity and low catalase levels which result in increased formation of free radicals and accelerated lipid peroxidation. Lipid peroxidation in turn results in damage to cellular membrane, disruption of ionic barrier, nucleic acid damage, and cell death.[10]

There are various antioxidant mechanisms against free radical damage, and Vitamin E is an important lipid-soluble chain-breaking antioxidant, preventing lipid peroxidation in membrane system.[11],[12] Vitamin E supplementation has been speculated to reduce lipid peroxidation in many studies. Hence, the present study was designed to investigate role of Vitamin E supplementation on free radical generation and lipid peroxidation in acute ALP poisoning in rats.

 Materials and Methods



Disease-free albino rats of both sexes from the Departmental Animal Room of Medical College Rohtak fed on “Gold Mohur rat feed” were selected for the study. During entire period of study, all the rats were caged separately. During the entire period of study, all the rats were handled as per the “Guiding Principles for Research Involving Animals and Human Beings” under the “Recommendations from the Declaration of Helsinki” and “Guiding Principles in the Care and Use of Animals” as approved by the Council of American Physiological Society.[13]

The present study was done in 30 disease-free albino rats weighing 125–250 g of both sexes to study the effect of acute ALP poisoning. The group for study on acute toxicity was further divided into three subgroups: A, B, and C. Each subgroup consisted of 10 rats.

Group A – Rats dosed with vehicle (ginni oil) only acting as control – 5 ml of prepared ginni oil was givenGroup B – Rats dosed with acute celphos poisoning – 5 ml “celphos mixture” (or 0.3 mg/g body weight)Group C –Rats dosed with acute celphos poisoning along with Vitamin E (1.5 mg Vitamin E/g body weight of rat.

The rats were administered the doses via an infant feeding tube no. 8 and blood was obtained by cardiac puncture 1 h after feeding the dose. Ginni oil acted as vehicle in our study for the administration of celphos and Vitamin E. The malondialdehyde (MDA) levels were estimated by modified method (thiobarbituric acid method) of Placer et al.[14] Superoxide levels were estimated by a slightly modified method as described by Baehner and Nathan (nitroblue tetrazolium [NBT] method).[15]

 Results



[Table 1] shows the serum MDA concentration in different groups of rats. MDA levels were found to be significantly high in the group that was administered celphos (Group B) as compared to the normal group (Group A) (17.48 + 3.17 μmol/L vs. 2.80 + 0.69 μmol/L, P = 0.000, respectively). In Group C, administration of Vitamin E resulted in decreasing the MDA level as compared to Group B (from 17.48 + 3.17 μmol/L to 14.53 + 0.71 μmol/L) (P = 0.025). The MDA levels in Group C still remained significantly high compared to the normal group.{Table 1}

NBT reduction was expressed as ΔOD/15 min/mL serum. NBT reduction was significantly increased in acute ALP poisoning group (Group B) as compared to the normal group (Group A) (0.45 + 0.06 vs. 0.19 + 0.08; P = 0.000) as shown in [Table 2] and [Figure 2]. Administration of Vitamin E to the rats of Group C resulted in significant decrease of NBT reduction compared to the Group B (P = 0.000).{Table 2}{Figure 2}

 Discussion



In the present study, NBT reduction was significantly increased in acute ALP poisoning group as compared to the normal group [0.45 + 0.06 vs. 0.19 + 0.08; P = 0.000; [Table 1]. Administration of Vitamin E to the rats of Group C resulted in significant decrease of NBT reduction compared to the Group B [P = 0.000, [Table 2] and [Figure 2].

ALP may induce oxidative stress leading to free radical generation and altered antioxidant status. PH3 acts at mitochondrial levels and generated free radical as is evident from the present study that NBT reduction is drastically increased. Furthermore, cellular injury due to PH3 induced free radical lipid peroxidation causing injury to the cell. This is evident from increased MDA levels in acute poisoning cases.[6] It is not clear in literature whether PH3-induced oxidation is the cause of PH3-induced mortality in most experimental models.[16]

Studies have proved that PH3-mediated inhibition of cytochrome c oxidase and other enzymes had led to the generation of superoxide radicals and cellular peroxides.[17] At cellular level, it is suggested to inhibit mitochondrial respiration and electron transport, the critical electrochemical link between respiration and phosphorylation in mitochondria.[2]

The present study reported that serum MDA concentration were significantly higher in the group who were administered celphos (Group B) as compared to the normal group (Group A) [Table 1]. In Group C, administration of Vitamin E resulted in decreasing the MDA level compared to Group B [P = 0.025, [Table 1] and [Figure 1]. The MDA levels in Group C still remained significantly high compared to the normal group and even after Vitamin E supplementation [Table 1].{Figure 1}

In the presence of ALP, cellular superoxide and peroxide radicals are generated, with subsequent cellular damage by lipid peroxidation. Increased lipid peroxidation resulting in raised MDA levels after ALP has been reported in literature.[18],[19] PH3 inhibit ADP uncoupler and ion-stimulated respiration due to the direct effect on electron transport, by interaction of PH3 with heme moiety of cytochrome oxidase.[1] Free radicals, MDA generated as a result of oxidative stress caused by aluminum toxicity. High levels of SOD and MDA seen in nonsurvivors suggested their direct relation to mortality, whereas the catalase has inverse relationship.[10] Chugh et al. have shown that celphos ingestion leads to high SOD and low catalase levels that result in increased formation of free radicals and accelerated lipid peroxidation.[10]

Recent reports on aluminum poisoning have been shown accelerated oxidative damage to biomolecules such as lipid, protein, and nucleic acids.[20] Ibegbu et al. in their experimental study on rats after supplementation of Vitamin E reported that Vitamin E reduces the toxic effect on the liver. Ability of Vitamin E to ameliorate or inhibit the action of phostoxin could be due to removing the ROS via very rapid electron transfer chain that inhibit lipid peroxidation. Vitamin E inhibits peroxidation of membrane lipids by scavenging lipid peroxyl radical with formation of tocopheroxyl radical.[21] Vitamin E counteracts harmful effects of aluminum not only by preventing free radical formation but also by favoring aluminum disposal,[22] therefore resulting in decreased MDA levels after Vitamin E supplementation in ALP.

Findings of the present study regarding reduction in MDA levels and NBT reduction (superoxide generation) following Vitamin E supplementation in rats give support to the concept that oxidative stress is induced by PH3 poisoning and possibly ALP toxicity causes disproportionate production of free radicals in tissue. Free radicals exerts toxic effects on various tissues via lipid peroxidation and weaken the antioxidant defenses, and findings of the present study clearly indicate that exogenous antioxidant agents such as Vitamin E in the treatment of acute ALP poisoning.[16]

Vitamin E may modulate mitochondrial production and levels of superoxide by preventing electron leakage, by mediating the superoxide generation systems directly, and/or by scavenging superoxide generated. By downregulating mitochondrial generation of superoxide and related reactive oxygen species, Vitamin E not only attenuates oxidative damage but also modulates the expression and activation of signal transduction pathways and other redox-sensitive biological modifiers.[23] The treatment with Vitamin E is effective in increasing both FcR- and CR3-mediated O2 production.[24]

More studies are required to study the role and of antioxidant agents to show the relationship between oxidative damage PH3-induced mortality.

 Conclusion



Findings of the present study showed that Vitamin E via its antioxidant action and anti-inflammatory effects has protective effect on PH3-induced toxicity in rats.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.

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