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DOI: 10.5958/j.2319-5886.2.2.


International Journal of Medical Research & Health Sciences


Volume 2 Issue 2 April - June


Copyright @2013

ISSN: 2319-5886

Received: 20 Mar 2013 Research article

Revised: 22 Mar 2013

Accepted: 26 Mar 2013

1 2

Dept. of ENT, L N Medical College & RC, Bhopal,India Dept. of ENT, Chirayu Medical College & Hospital-Bhopal, India

*Corresponding author email: drpatelent@yahoo.com


Background: Globally Oral Cancer is the sixth most common cause death with India accounts for 86% of the worlds oral cancer cases. Chronic tobacco quid consumption often results in a progressive premalignant condition called Oral Sub mucous Fibrosis (OSMF) whose malignant transformation rate of is around 7.6%. Free radicals released during the metabolism of tobacco and Areca nut my involved in the initiation and propagation of mucosal fibrosis. Objective: the objective of the present study is to measure antioxidant enzymes and lipid peroxidation levels in OSMF to assess oxidative stress like environment in OSMF patients. Materials and methods: for this study we invited 38 newly diagnosed OSMF patients both male and female consuming tobacco in the form of quid and the same number of age matched healthy non tobacco consuming were selected as a control group. In both groups plasma superoxide dismutase, Glutathione peroxidase, catalase levels and lipid peroxidation rate was measured. Results and conclusion: we observed very low antioxidant enzyme levels in OSMF patients when compared with healthy controls (P<0.01) and at the same time also observed very high lipid peroxidation rate in the study population (P<0.01) compare to control group indicating prevalence of oxidative stress like environment in tobacco consuming population, which might play a vital role in the initiation and propagation of various precancerous conditions like OSMF. Keywords: Oral submucous fibrosis; Antioxidant enzymes, non smoking tobacco consumption

Cancer is one of the most common causes of mortality and morbidity today, with more than 10 million new cases and more than 6 million deaths each year worldwide. Global Oral Cancer is the sixth most common cause of cancer related death1. India accounts for 86% of the worlds oral cancer cases, says the study conducted by the National Institute of Public Health in

February 2011. Oral cancer accounts for approximately 3040% of all cancers in India. The strong association between oral cancers with tobacco use is well established. Many epidemiological studies showed the risk of developing oral cancer is five to nine times greater in smokers than nonsmokers. In addition to smoking tobacco chewing has been associated

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Int J Med Res Heatlh Sci. 2013;2(2):229-232

with increased risk of oral cancer2. In the Indian subcontinent the chronic use of betel quid has been strongly associated with increased risk for oral cancer3. The quid typically consists of betel leaf, Areca nut, tobacco pieces, and lime. In presence of lime, Areca nut releases high concentration of alkaloids. Chronic tobacco quid consumption often results in a progressive premalignant condition called Oral Sub mucous Fibrosis (OSMF). From the study done by Pillai P N et al4 observed that the malignant transformation rate of OSMF is around 7.6%. The major etiological factor for the development of OSMF on chronic consumption of Areca quid may be due high concentration of alkaloid and the interaction of these alkaloids with free radicals liberated from tobacco with the oral mucosa. Thus the aim of this present study is to estimate plasma antioxidant enzymes like Super oxide dismutase (SOD), glutathione peroxidase (GSH-Px), catalase (CAT) and lipid peroxidation in OSMF patients evaluate oxidative stress levels in OSMF and its role in initiation and development of this disease.

was taken, transferred to a heparinised test tube and centrifuged at 4C and the plasma was stored at 80oC till the biochemical investigations were done. In both control and OSMF groups we estimated antioxidant enzymes like SOD5, GSHPx6 and Catalase7 and the lipid peroxidation rate was determined by estimating Malondialdehyde which is also called thiobarbituric acid reactive substances (TBARS)8. SOD was estimated auto oxidation method by adding 0.5ml of plasma to 1.8mM epinephrine and 0.6mM EDTA and the increase in absorbance was read at 480nm. GSHPx was measured by DTNB method .03 ml of hemolysate to .04% DTNB (Dithiobis nitro benzoic acid) and 1.215mM H2O2 and the change in abosrbance was taken at 412nm. Plasma catalase was assessed by Goths methods adding 0.2ml of plasma to 60mmol/l H2O2 and 32.4nmol/L ammonium molybdate and change in absorbance was measured at 405nm. Statistical Analysis A Student t test was applied to assess the statistical difference of the above said biological parameters between OSMF patients and control group. P <0.05 was considered as significant. Statistical analysis was done by SPSS 18 version. RESULTS Enzymatic antioxidant defense status in OSMF was assessed by measuring plasma Superoxide Dismutase, Catalase and Glutathione Peroxidase. One of the prominent adverse effects of free radicals was estimated by measuring plasma Malondialdehyde (MDA). The plasma levels of these antioxidant enzymes and MDA were compared with control group (Table I). We observed very low levels of antioxidant enzymes, and very high MDA in OSMF when compared to control group (p < 0.01).

This study was approved by the Institutional Ethical Committee, and written consent was taken from every participant. Study group comprises 38 newly diagnosed both male and female OSMF patients of age between 25 to 45 years who have not received any previous treatment or on any antioxidant therapy and were confirmed after the detailed case history and histopathological confirmation. For the control group same number of age and sex matched healthy individuals who are non tobacco and areca quid consumers, who were not suffering from any systemic illness, were selected. From both control and study group 5 ml venous blood


Teklal et al.,

Int J Med Res Heatlh Sci. 2013;2(2):229-232

Table-1: Antioxidant enzyme and lipid peroxidation levels in control and OSMF patients. Parameters Control n=38 OSMF group n=38 Malondialdehyde (MDA) nmol/ml 1.04 0.14 3.1 0.58* Superoxide dismutase (SOD) U/g Hb 1104 126 527 84* Glutathione peroxidase (GSH-Px) U/g Hb 91 6.3 48 4.2* Catalase (CAT) KU/L 24 5.1 14 3.5* Results were presented as mean + SD. *P <0.01

In the present study by assessing the plasma levels of antioxidant enzymes SOD, CAT, GSHPX levels and MDA we observed the prevalence of high oxidative stress like environment in OSMF because of release of high free radicals and reactive oxygen species from smokeless tobacco consumption in the form of quid. The tar phase of tobacco contains several relatively stable free radicals; of which the principle radical identified was a quinone/hydroquinone (Q/QH2) complex held in the tarry matrix9. This Q/QH2 polymer is an active redox system that is capable of reducing molecular oxygen to produce superoxide, eventually leading to hydrogen peroxide and hydroxyl radicals. Along with this the high copper content of areca a content of quid also contributes in the generation of free radicals by Fenton and Haber - Weiss reactions10. H2O2 + Cu (I) Cu (II) + OH + OH(I) Fenton Reaction H2O2 + O2O2 + OH + OH- (II) Haber-Weiss Reaction Through these mechanisms excessively generated free radicals in tobacco quid consumption were quenched by bodys first line of antioxidant mechanism that is chain breaking mechanism through antioxidant enzymes like SOD, CAT and GSH-Px. This might be the reason for decreased concentration of these enzymes which were observed in the study population. The same decrease in antioxidant enzymes in tobacco consuming OSMF were observed by many researchers11, 12. In the present

study we also observed higher levels of MDA in tobacco consuming OSMF patients. This may be due to adverse effects of hydroxyl radical OHradical which was produced by the Q/QH2 polymer of tobacco. OH- oxidizes double bonds of polyunsaturated fatty acids (PUFA) of cell membrane, thus produces the end product called Malondialdehyde (MDA). There is substantial evidence that the OH- radical generated from copper through the Fenton reaction can destruct tissue by initiation and propagation of lipid peroxidation by absorbing hydrogen from PUFA of cell membrane to produce MDA. Our results are positively correlated with other researchers also11, 13. The oxidative stress like environment which prevails in this group might render as an individual more susceptible to damage, which might have led to the development of sub mucous fibrosis. The decrease in antioxidant enzymes in this study might be due to consumption of these enzymes in quenching of high levels of free radicals which are generated due tobacco metabolism.

From this study it was concluded that in OSMF patients very low levels of antioxidant enzymes and higher lipid peroxidation in Areca quid consuming OSMF patients. Areca and tobacco metabolism might produce high levels of free radicals which might play an important role in the initiation and propagation of fibrosis of the oral mucosa.


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Teklal et al.,

Int J Med Res Heatlh Sci. 2013;2(2):229-232