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ORIGINAL ARTICLE
Year : 2017  |  Volume : 44  |  Issue : 3  |  Page : 137-139

Estimation of serum malondialdehyde as a marker of lipid peroxidation in medical students undergoing examination-induced psychological stress


1 Obesity Research Laboratory, Interactive Research Schoolfor Health Affairs, Pune, Maharashtra, India
2 Department of Sharir Kriya, College of Ayurveda, Bharati Vidyapeeth Deemed University, Pune, Maharashtra, India

Date of Web Publication14-Feb-2018

Correspondence Address:
Supriya Bhalerao
Obesity Research Laboratory Interactive Research School for Health Affairs, Bharati Vidypeeth Deemed University, Pune - 411 043, Maharashtra
India
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/jss.JSS_13_17

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  Abstract 

Background: When oxidant compounds target lipids, they can initiate the lipid peroxidation process, a chain reaction that produces multiple breakdown molecules, such as Malondialdehyde (MDA). Psychological stress is reported to induce enhancement of lipid peroxidation in the brain. Objective: The present study was therefore planned to evaluate whether there is increase in oxidative stress in medical students undergoing examination induced psychological stress. Materials and Method: Institutional Ethics Committee permission and seventy nine students of either sex between the age group 19 to 21 years. Students studying in first year of medical college were recruited in the study. All students signed the written informed consent before collecting blood samples. 5 ml of blood was collected from each student in a plain vaccutainer just before appearing for viva examination. Serum was separated immediately by centrifugation and MDA was estimated using (TBA) thiobarbituric acid method. Result: We observed that in 24 individuals lipid peroxidation was higher (>3 nmol/ml) than normal level (0-3 nmol/ml). The number of female students showing higher values of MDA was higher than male students with similar values of MDA (p<0.05). Conclusion: Our study thus highlights the relation between lipid peroxidation and psychological stress and also the sex wise difference in the stress levels.

Keywords: Lipid peroxidation, malondialdehyde, reactive oxygen species, stress


How to cite this article:
Banjare J, Salunke M, Indapurkar K, Ghate U, Bhalerao S. Estimation of serum malondialdehyde as a marker of lipid peroxidation in medical students undergoing examination-induced psychological stress. J Sci Soc 2017;44:137-9

How to cite this URL:
Banjare J, Salunke M, Indapurkar K, Ghate U, Bhalerao S. Estimation of serum malondialdehyde as a marker of lipid peroxidation in medical students undergoing examination-induced psychological stress. J Sci Soc [serial online] 2017 [cited 2018 May 23];44:137-9. Available from: http://www.jscisociety.com/text.asp?2017/44/3/137/225497


  Introduction Top


Oxidative stress is the damage to cells caused by oxidation, which causes a large increase in the cellular reduction potential. It causes destruction of cells by the production of reactive oxygen species (ROS).[1] ROS are chemically reactive molecules containing oxygen. In low levels, they get countered by the cell antioxidants. However, severe levels of oxidative stress cause damage to the cells. This damage causes adenosine triphosphate depletion, leading to uncontrolled apoptotic death.

Lipid peroxidation (autooxidation) is a chain reaction providing a continuous supply of free radicals that initiates further peroxidation in the lipid-rich membrane of lipoproteins by ROS such as hydroxyl radical nitric oxide radical superoxide radical, and ROO (Peroxyl radical) resulting in generation of lipid peroxidation products like malondialdehyde (MDA).[2] MDA (bis–diethylacetal) CHO-CH2-CHO, is a dicarbonyl, with a molecular weight of 72 Daltons, formed as a secondary product of endoperoxide during endogenous biosynthesis of prostaglandins and leukotrienes from arachidonic acid and other PUFA. Increased levels of the lipid peroxidation product MDA play a very important role in the pathogenesis of various diseases.[3]

Currently, research dedicated to the psychobehavioral modulation of immune function called psychoneuroimmunology have shown the stress-induced immune impairment.[4] The immune dysfunction leads to inflammation and damages cell turning to for lipid peroxidation. Studies have reported that oxidative damage is highly correlated with neutrophil activity during examinations in controlled demographic and health behaviors.[5] In another study, results suggest that during university examinations students are under increased oxidative stress.[6]

In the present study, we, therefore, estimated serum MDA levels in medical students undergoing examination just before appearing for viva.


  Methodology Top


After obtaining the Institutional Ethics Committee permission, seventy-nine students of either sex between the age group 19–21 years and studying in the 1st year of medical college were recruited in the study. A written informed consent from all the students was obtained before collecting blood samples from them.

A volume of 5 ml of blood sample was collected from each student in a plain vacutainer just before appearing for viva as a part of the internal examination. Serum was separated immediately by centrifugation and MDA was estimated using the thiobarbituric acid (TBA) method. In short, MDA present in serum is precipitated with weak trichloroacetic acid and boiled for 30 min with 0.67% TBA in 2M sodium sulfate reagent in acidic medium (0.05M H2SO4) which results in hydrolysis of C = N bonds of conjugated Schiff's base of MDA protein adduct. The liberated MDA couples with TBA to form a pink 1:2 (MDA: TBA) adduct. This chromogen is extracted with n-Butanol on cooling and absorbance measured at 532 nm.

The results are presented as mean ± standard deviation MDA levels as well as a number of participants (frequency) in a specified range of MDA.


  Results Top


We estimated the MDA levels in 79 medical students undergoing examination stress. Of these, 48 were female and 31 were male students.

The mean MDA levels in these participants were found to be 2.82 ± 2.49 nmol/L. Further, 55 study participants showed MDA levels in the range 0–3 nmol/L, whereas 24 participants showed MDA levels >3 nmol/L. The distribution of the MDA levels thus showed a binomial distribution. The data are summarized in [Table 1] and [Figure 1].
Table 1: Serum malondialdehyde (nmol/ml) level of participants

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Figure 1: Serum malondialdehyde level in the study participants

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When the data were analyzed with respect to sex, the mean levels in male participants were 2.46 ± 2.40 nmol/L, whereas the levels in female participants were 3.06 ± 2.53 nmol/L. Although the values were higher in females, they were not statistically significant.

Further, 16 female participants showed higher levels of MDA. This number was significantly higher than the male participants (n = 6) with similar MDA levels (P < 0.05).


  Discussion Top


In the present study, we evaluated MDA [marker of oxidative stress in various health complications[7],[8] levels in medical students undergoing examination-induced psychological stress. We found that out of 79 participants, 24 participants had higher levels of MDA, whereas 55 participants showed normal levels. This finding indicates that in spite of the similar exposure, i.e., examination induced psychological stress; there is difference in the oxidative stress response in the individuals.[6] The intra-individual variation we observed may be due to polymorphisms in the genes regulating anti-oxidant mechanism such as hOGG1 Ser326Cys, 8-oxoguanine glycosylase 1Glutathione S-transferases M1, T1, and P1 polymorphisms, etc.[9]

This study also revealed that there is sex-wise difference in the levels of MDA. Female students demonstrated higher oxidative stress indicating higher level of psychological stress. This confirms the higher psychological stress in females well reported in earlier studies.[10]

Oxidative stress has recently been linked with a number of chronic and inflammatory diseases. Free radicals within physiological concentration regulate cognitive functions.[11] However, oxidative stress resulting from excessive generation of reactive species compared with antioxidant defense capacity can cause lipid peroxidation and these oxidant products have a central role in induction of psychiatric dysfunction that indicates links between free radicals and neurological illness.[12]

The study however has certain limitations. It would have been interesting to study the perceived stress levels in the students and associate them with the MDA levels. More emphasis on history and clinical data would have provided idea about potential confounders in the study. In spite of these limitations, our study adequately highlights the increased oxidative stress in individuals undergoing psychological stress. In future, a well-planned study considering the above mentioned limitations can be conducted which can be expanded to explore mechanism of psychological stress-induced oxidative stress.


  Conclusion Top


From the above study it was evident that Malondialdehyde (MDA) level is important biomarkers of oxidative stress due to increased lipid peroxidation which is a risk factor for various mental illness including stress, depression, Parkinson's disease etc.

Acknowledgment

We wish to express our gratitude to all students who participated in this study and to the Dr. Kavita Indapurkar and Dr. Umesh Ghate from Department of Sharirkriya, College of Ayurveda, Bharati Vidyapeeth Deemed University, Pune, (MH) for their kind support and providing assistance to perform the study at same place. This study was done in collaboration of Interactive Research School for Health Affairs and college of Ayurveda, Bharati Vidyapeeth Deemed University, Pune, (MH).

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.

 
  References Top

1.
Yin H, Xu L, Porter NA. Free radical lipid peroxidation: Mechanisms and analysis. Chem Rev 2011;111:5944-72.  Back to cited text no. 1
    
2.
Devasagayam TP, Tilak JC, Boloor KK, Sane KS, Ghaskadbi SS, Lele RD, et al. Free radicals and antioxidants in human health:Current status and future prospects. J Assoc Physicians India 2004;52:794-804.  Back to cited text no. 2
    
3.
Del Rio D, Stewart AJ, Pellegrini N. A review of recent studies on malondialdehyde as toxic molecule and biological marker of oxidative stress. Nutr Metab Cardiovasc Dis 2005;15:316-28.  Back to cited text no. 3
    
4.
Glaser R, Kiecolt-Glaser JK. Stress-induced immune dysfunction: Implications for health. Nat Rev Immunol 2005;5:243-51.  Back to cited text no. 4
    
5.
Judith Eggenberger C. Infl ammation and Oxidative Damage During Exam Stress. PhD Diss., University of Pittsburgh; 2006.  Back to cited text no. 5
    
6.
Sivonová M, Zitnanová I, Hlincíková L, Skodácek I, Trebatická J, Duracková Z, et al. Oxidative stress in university students duringexaminations. Stress 2004;7:183-8.  Back to cited text no. 6
    
7.
Markesbery WR. Oxidative stress hypothesis in Alzheimer's disease. Free Radic Biol Med 1997;23:134-47.  Back to cited text no. 7
    
8.
Nagamani M. Lipid peroxidation product as a marker of oxidative stress in psoriasis – A case control study in North Coastal Andhra Pradesh. J Dent Med Sci 2015;14:18-20.  Back to cited text no. 8
    
9.
Jenner P. Oxidative stress in Parkinson's disease. Ann Neurol 2003;53 Suppl 3:S26-36.  Back to cited text no. 9
    
10.
Sayre LM, Smith MA, Perry G. Chemistry and biochemistry ofoxidative stress in neurodegenerative disease. Curr Med Chem 2001;8:721-38.  Back to cited text no. 10
    
11.
Babusikova E, Evinova A, Hatok J, Dobrota D, Jurecekova J. Oxidative changes and possible effects of polymorphism of antioxidant enzymes in neurodegenerative disease. InTech In Neurodegenerative Diseases 2013.  Back to cited text no. 11
    
12.
Taylor SE, Klein LC, Lewis BP, Gruenewald TL, Gurung RA, Updegraff JA, et al. Biobehavioral responses to stress infemales: Tend-and-befriend, not fi ght-or-fl ight. Psychol Rev 2000;107:411-29.  Back to cited text no. 12
    


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