Study of Docosahexaenoic Acid and Eicosapentanoic Acid Effects on Some Biochemical Parameters in Epileptic patients
Dania E. Ibrahim1, Wisam Kadhum H. Alhashemi2, Sajid Ibrahim Alhussaini3
1Department of Chemistry, College of Science, Al-Nahrain University, Baghdad-Iraq
2Department of Chemistry, College of Science, Al-Nahrain University, Baghdad-Iraq
3Consultant of Neurology, Private Nursing Home Hospital, Baghdad Teaching Hospital in Department of Medicine City, Board of Neurology, Medical Training Center, Baghdad-Iraq
*Corresponding Author E-mail: wisamchem1@yahoo.com
ABSTRACT:
Epilepsy is a worldwide disease characterized by huge changes in several biochemical parameters due to seizure, the main symptoms of the disease. The aim of this study was to assess some biochemical parameters in epileptic patient. This study involves 31 patients with 31 healthy group of apparently healthy subjects as a control group for each one of both groups these parameters were assessed: {8-hydroxydeoxy guanosine (8-OHdG), Malondialdehyde (MDA), Prolactin, Lactate dehydrogenase (LDH), Albumin, Total serum bilirubin (TSB), Alkaline phosphatase (ALP), Aspartate aminotransferase (AST), Alanine-aminotransferase (ALT) and Creatine phospho kinase (CPK)} in serum of them. 8-OHdG concentrations was increase significantly (P> 0.05) in epileptic patients in compare to control group while MDA show nonsignificant change, the seizure was characterized by increasing of oxidative stress. Situation manifested by its effects, mainly, on nucleic acid. TSB was significantly decreased (P> 0.05) may be due to its function as endogenous antioxidant. albumin did not show any significant difference. CPK has significant increasing (P> 0.05) which be due to the seizure nature, that need high energy production which stimulate the reactions of energy production which lead to upregulate that enzyme. LDH failed to show any significant difference may be due to fast remediation. When comparison the epileptic patients groups during (0,1 and 2) month of supplementation with EPA and DHA with control subjects. the results were as follow: significantly decrease in 8-HdG concentration level (17.9888±4.30645) ng/ml with (P> 0.05) in 0 month, (14.8998±3.89157) ng/ml with (P> 0.05) in 1 month and (8.5775±2.15075) ng/ml with (P <0.05) in 2 month at the comparison with control group (7.1606±1.23027) ng/ml. while show MDA show non-significant difference in comparison control with the epileptic patients in (0,1 and 2) month of supplementation with EPA and DHA with control subjects. Bilirubin concentration level show significant decrease (0.4693±0.04143) mg/dl with (P> 0.05) in 0 month, (0.4096±0.02873) ) ng/ml with (P> 0.05) in 1 month and (0.3983±0.04033) ng/ml with (P <0.05) in 2 month at the comparison with control group (0.6603 ±0.06761) ng/ml while Albumin, GOT, GPT and ALK show non-significant difference in comparison control with the epileptic patients in (0,1 and 2) month of supplementation with EPA and DHA with control subjects. In CPK, LDH and Prolactin show non-significant difference in comparison control with the epileptic patients in (0,1 and 2) month of supplementation with EPA and DHA with control subjects.
KEYWORDS: Epilepsy, Omega3, EPA, DHA, 8-hydroxydeoxy guanosine, Malondialdehyde, Creatine phospho kinase, oxidative stress.
INTRODUCTION:
Epilepsy is one of the most common neurologic problems worldwide. Approximately 1% to 2% of the population suffers from epilepsy, making it the second most common neurologic disorder (after stroke)[1]. It has been estimated that about 7%–8% of the population experiences at least 1 epileptic seizure during their lifetimes[2]. A seizure is a clinical appearance, producing from a short episode of abnormal undue or concurrent neuronal activity in the brain. The tendency to have recurrent unprovoked seizures is called ‘Epilepsy’. Two unprovoked seizures were demands by epilepsy definition, separated by major than 24 hours[3] To function normally, an ongoing equilibrium between excitation and inhibition was achieved in the brain, to still responsive to the surroundings without continuous unrestrained spontaneous action. It is possible that many seizures produce from an inequality between this excitation and inhibition.[4] Docosahexaenoic acid (DHA) is the N-3 PUFA in the brain; it is including in the organizing of neuronal function by several various mechanisms, like regulation of neurotransmitter release, interaction with ion channels, the signals of neurotransmitter and nuclear-receptor–mediated transcription of genes and composes of biologically efficient oxygenated derivatives. modulation of membrane biophysical properties by incorporate into the phospholipids of cell membrane, the membrane fluidity altering in this way, also altering the structure of the lipid rafts, and then changing the potassium current, and also change the expression level of various potassium channels[5][6]. The High concentrations of omega-3 are found especially at the synaptic level, in order to have a flexible and a maximum potential of the neuronal functionality[7]. One of PUFA source is the elongation process of alpha-linolenic acid (ALA) and linoleic acid (LA) that place in the blood-brain barrier (BBB) and in almost all cells in the brain. The other of PUFA source is from the diet itself. How almost all of the PUFA do not known the capable to perform the passing, when the fatty acids that drive from the diet have to pass the blood brain barrier.
Oxidative stress and mitochondrial dysfunction happened as a production of long-time epileptic seizure and the seizure-induced brain damage may provide. After all, the epileptogenic is the acute differences in seizure-induced free radical construction and mitochondrial dysfunction[8]. The end product of hydroxylation of guanine is 8-hydroxydeoxy guanosine (8-OHdG). Once formed, its level will raise in the place of DNA injury and freed to be create in the plasma and the urine, raised oxidative stress effected by seizure action raises the occurrence of oxidative DNA damage in all brain domains, containing hippocampal neurons [9] Epilepsy effects on MDA level which stimulating of Glutamate for the N-methyl-D-aspartate (NMDA) receptors, leading to raised excitotoxicity and cerebral hypoxia, that produces from raised inflow of calcium by way of voltage-gated and NMDA-dependent ion channels leading to raise in ROS and oxidative stress[9] Epilepsy effected on Liver function tests (AST, ALT and ALP) by significant elevation of their serum levels in epileptic patients[10][11]. Albumin view as a bigger antioxidant in plasma[13] that has actions related to ligand-binding capacities[14]. Total bilirubin is another effective antioxidant that could prevent lipid oxidation and other kinds of oxidation[13] and effected by epilepsy disease[15][16]. The Serum albumin can do as an effective pro-synaptogenic signaling molecule, proposing that control of Blood Brain Barrier integrity may promot regulated synaptogenesis in damaged tissue[17]. The epilepsy has a effective role with CPK serum level which the cellular constituent may be a target for free radical injury, the restriction of some target such as Na+, K+-ATPase may act as a serious function in the epilepsy that caused hyperexcitability Na+, K+-ATPase that is a membrane bound enzyme well-known to act as a serious function in cellular ionic gradient support and particularly delicate to reactive species[16]. LDH enzyme has extracellular activity raises under the action of oxidative stress[18]. Increasing of sex hormone prolactin, Disturbance of central and/or peripheral control of hypothalamic-pituitary-gonadal axis and alteration of central neurotransmitters (GABA, glutamate and serotonin) by epileptic discharges. The release of neurotransmitters including γ-aminobutyric acid (GABA), glutamate, opioids and serotonin also contribute to the release of hypothalamic and pituitary hormones and intact sexual function[19].
MATERIALS AND METHODS:
This study involved 31 patients where they have epilepsy disease (20 males 11 females), age between (15-64) years. The samples were collected from privet clinic in the period from September 2017 to March 2018, all patients were diagnosed by Consultant Physician in Neurology. Thirty-one volunteers were also included in this study as a control group (19 males and 12 females), age between (15-53) years. The parameters used in this study were; Serum MDA (kit supplied by SHANGHAI YEHUA Biological Technology), Serum Albumin (kit supplied by BIOLABO), Serum total Bilirubin (kit supplied by BIOLABO), serum 8-OHdG (kit supplied by SHANGHAI YEHUA Biological Technology), serum Prolactin (kit supplied by Monobind lnc.), serum GOT (kit supplied by BIOLABO), serum GPT (kit supplied by BIOLABO), serum ALP (kit supplied by BIOLABO), serum LDH (kit supplied by HUMAN) and serum CPK (kit supplied by HUMAN).
Statistics analysis:
In this study all statistical analysis used SPSS program. To descriptive analysis was used to show the mean and standard error mean (SEM) by using Student's T-test. The figures were done by used Microsoft Excel program. Difference consider significantly at (p<0.05)[20] [[21] [22]. Post Hoc tests were used for multiple comparisons between groups. The probability P≤0 .05 was considered as and significant P>0.05 non-significant.
RESULTS AND DISCUSSION:
Number and percentage (according to gender) of subjects who's involved in this study are given in.
Table 1: Distribution of subjects according to the healthy status and gender.
|
Study group |
Healthy control |
Epileptic patients |
|||
|
N |
% |
N |
% |
||
|
Gender |
19 |
61% |
20 |
65% |
|
|
Female |
12 |
39% |
11 |
35% |
|
|
Total |
31 |
100% |
31 |
100% |
|
|
BMI (kg/m2) (Mean ± SEM) |
24.84±0.92 |
26.44±1.57 |
|||
|
Age in year |
15 – 53 |
15 - 64 |
|||
|
Duration |
- |
From birth to 64 years |
|||
The mean (±SEM) values of serum 8-HdG levels in addition to size number and P-Value in control and epileptic patients in (0, 1 and 2) month of supplementation with EPA and DHA with control subjects are listed in table (2) and figure (1).
Table 2: Comparison of the 8-HdG concentrations between the epileptic patients in (0, 1 and 2) months of supplementation with EPA and DHA with control subjects
|
Subjects/month |
N |
Mean ± SEM (ng/ml) |
P-Value |
|
Normal |
31 |
|
|
|
Patient 0 |
20 |
17.9888 ±4.30645 |
.007* |
|
Patient 1 |
20 |
.050* |
|
|
Patient 2 |
19 |
.721 |
*High significant with P<0.05. 8-HdG: 8- Hydroxydeoxy Guanosine, EPA: Eicosapentanoic Acid, DHA: Docosahexaenoic Acid, SEM: Standard Error Mean.
Figure 1 : Comparison of the 8-HdG concentrations between the epileptic patients in (0, 1 and 2) month of supplementation with EPA and DHA with control subjects
It’s obvious from results in table (2) in above that 8-HdG as oxidative stress marker decrease in its concentration along period of supplementation reaching to nonsignificant difference in compare with control group which may be reflect the omega 3 effect as modulating agent for oxidative stress process.
The mean (±SEM) values of serum bilirubin levels in addition to size number and P-Value in control and epileptic patients in (0,1and2) month of supplementation with EPA and DHA with control subjects are listed in table (3) and figure (2).
Table Error! No text of specified style in document.: Comparison of the bilirubin test concentrations between the epileptic patients in (0, 1 and 2) months of supplementation with EPA and DHA with control subjects.
|
Subjects/month |
N |
Mean ± SEM (mg/dl) |
P-Value |
|
Normal |
30 |
|
|
|
Patient 0 |
28 |
.005* |
|
|
Patient 1 |
25 |
.000* |
|
|
Patient 2 |
18 |
.001* |
*High significant with P<0.05, 8-HdG: 8- Hydroxydeoxy Guanosine, EPA: Eicosapentanoic Acid, DHA: Docosahexaenoic Acid, SEM: Standard Error Mean.
Figure 2: Comparison of the bilirubin test concentrations between the epileptic patients in (0, 1 and 2) month of supplementation with EPA and DHA with control subjects.
Bilirubin is significant differences effect with omega3 fatty acid, which omega 3 fatty acid don’t effect on bilirubin may be due to that bilirubin powerful antioxidant that is highly sensitive to any oxidizing agent.
Table 4: Number of seizure relationship between frequency of seizure and supplements of Omega3.
|
Duration of (N) |
Controlled |
Partly controlled |
Uncontrolled |
|||
|
Percentage |
No. of subjects |
Percentage |
No. of subjects |
Percentage |
No. of subjects |
|
|
Before supplementation (31) |
0% |
0 |
45% |
14 |
54% |
17 |
|
After 1 month (28) |
71% |
20 |
7% |
2 |
21% |
6 |
|
After 2 months (20) |
65% |
13 |
20% |
4 |
15% |
3 |
*N = number of cases involved, Controlled: zero seizure per month, Partly controlled: 1 seizure per month, Uncontrolled: 2 and more seizure per month.
This study involved 31 patients depending on the frequency of seizure: zero seizure per month mean controlled type, 1 seizure per month mean partly controlled type and 2 and more seizure per month mean uncontrolled type. There is (54% 17 of 31 subjects) uncontrolled, (54% 14 of 31subjects) partly controlled and (0% 0 of 31 subjects) uncontrolled in zero month. After 1 month the percentage change significantly with (21% 6 of 28 subjects) uncontrolled, (7% 2 of 28 subjects) partly controlled and (71% 20 of 28) controlled. After 2 months the percentage change significantly with (15% 3 of 20 subjects) uncontrolled, (20% 4 of 20 subjects) partly controlled and (65% 13 of 20) controlled. These results give us obvious indication of beneficial of omega3 supplementation which may be due to its action as anti-inflammatory and [reduce oxidative stress and morphological effect (such as change channels)].
According to Chi-Square test there are significant decrease in uncontrolled epileptic patients (54% to 15% from total patients) in compare to significant increase in controlled epileptic patients (0% to 65%) along omega3 supplementation.
CONCLUSION:
The effect of omega3 on cell functions obvious through significant changes in many biochemical parameters through its action has anti-inflammatory effects and also may be as pharmacological change due to replacements of other fatty acids in cell. Omega 3 supplementation have beneficial effects on biochemical parameters and symptoms of epilepsy.
Oxidative stress biomarker (MDA and 8-HdG) was a clear indicator to distinguish epileptic patients from control healthy group and to know how useful of omega3 supplementation in epileptic patients which refer to the DNA in oxidative stress while MDA was failed to show how useful of omega supplementation. In contrast the liver function parameter was failed to show any effect between epileptic patients and control before and after omega3 supplementation, these was occurred may be due to of the liver function parameters not effect neither in this disease nor its medicines substantially, because most of these patients (that were selected) supplemented the epileptic medicines for at least 5 years, except for bilirubin was significantly changes, may be due to its antioxidants properties. While functional enzymes (CPK) and metabolic enzymes (LDH) that related to the epileptic patients were made the distinction before and after supplementation, this pointing to good indicator for differentiate and followed supplementation. Epileptic patients have significant difference before and after supplementation in oxidative stress biomarker (8-HdG) and in Bilirubin. From all the above we can conclude the beneficial effects of DHA and EPA supplementation on biochemical parameters and symptoms of elliptic patients.
ACKNOWLEDGEMENTS:
The authors would like to thank Al- Nahrain University, and Al-Zafaran private lab. In AL-Zafraniya city.
REFERENCES:
1 C. E. Stafstrom, "The Pathophysiology of Epileptic Seizures: A Primer For Pediatricians," Back to Basics, vol. 19, p. 342, 2018.
2 J.P. Karis, ACR Appropriateness Criteria, "Expert Panel on Neurologic Imaging," AJNR American Journal of Neuroradiology , vol. 29, p. 1222, 2008.
3 Hae Won Shin, Valerie Jewells, Eldad Hadar, and et al, "Review of Epilepsy - Etiology, Diagnostic Evaluation and Treatment," International Journal of Neurorehabilitation, vol. 1, p. 1, 2014.
4 Stephen L. Hauser, Dennis L. Kasper, Anthony S. Fauci, and et al, "Harrison’s neurology in clinical medicine," ketabpezeshki, p. 297, 2017.
5 Kazuhiro Tanaka , Akhlaq A. Farooqui , Nikhat J. Siddiqi, and et al, "Effects of Docosahexaenoic Acid on Neurotransmission," Biomolecules and Therapeutics, vol. 20, pp. 152-157, 2012.
6 M. Elizabeth Sublette, Hanga C. Galfalvy, and Joseph R. Hibbeln, "Polyunsaturated fatty acid associations with dopaminergic indices in major depressive disorder," International Journal of Neuropsychopharmacology, vol. 17, p. 383–391, 2014.
7 Oana Tarţa-Arsene, Mădălina Leanca, Alice Dică, and et al, "Dietary Omega-3 Fatty acids Supplementation for," Farmacia, vol. 65, pp. 550-556, 2017.
8 Patil CD, Ahire YS, Pathade PA, Pathade VV, and et al, "Free Radicals, Epilepsy and Anti-oxidant: An Overview," IRJP International Research Journal of Pharmacy, vol. 2, p. 3, 2011.
9 Basel A. Abdel-Wahab, Ibrahim A. Shaikh, Masood M. Khateeb, and et al, "Omega 3 polyunsaturated fatty acids enhance the protective effect of levetiracetam against seizures, cognitive impairment and hippocampal oxidative DNA damage in young kindled rats," Elsevier, p. 111, 2015.
10 Shahabeddin Rezaei, Mona Kavoosi, and Reza Shervin Badv, "The Influence of Ketogenic Diet on Liver Function in Children and Adolescents with Intractable Epilepsy," J Compr Ped, p. 2, 2017.
11 A.M. El-Mowafy, M.A. Abdel-Dayem, A. Abdel-Aziz, and et al, Eicosapentaenoic acid ablates valproate-induced liver oxidative stress and cellular derangement without altering its clearance rate: Dynamic synergy and therapeutic utility. Elsevier, vol. 1811, pp. 460-467, 2011.
12 S. S. a. J. S. Pramod J.. Role of free radicals and antioxidants in human health and disease. International Journal of Current Research and Review, vol. 19, p. 14, 2013.
13 B. A., "Bilirubin as savior of biological system," Int. J. Curr. Res. Med. Sci, vol. 2, pp. 21-27, 2017.
14 a. D. B. Bourdon E., "The importance of proteins in defense against oxidation," Antioxidants and Redox Signaling, vol. 2, pp. 293-311, 2001.
15 Becerir C, Kılıç İ, Sahin Ö, and et al, "The protective effect of docosahexaenoic acid on the bilirubin neurotoxicity, Enzyme Inhibition and Medicinal Chemistry,," Informa UK , p. 6, 2013.
16 Mathew George, Lincy Joseph and Preethi Christina Jose, "A review article on assessing the effect of antiepileptics," The Pharma Innovation Journal, p. 13, 2016.
17 L. L. K. a. e. a. Itai Weissberg, "Albumin induces excitatory synaptogenesis through astrocytic TGF-β/ALK5 signaling in a model of acquired epilepsy following blood-brain barrier dysfunction," Elsevier Neurobiology of Disease, vol. 78, pp. 115-125, 2015.
18 Predrag Jovanović, Lepša Žorić, Ivan Stefanović, and et al, "Lactate Dehydrogenase And Oxidative stress Activity in Primary open-angle Glaucoma aqueous humour," Bosnian Journal of Basic Medical Sciences, vol. 10(1), p. 86, 2010.
19 Sherifa A. Hamed, "The effect of epilepsy and antiepileptic drugs on sexual, reproductive and gonadal health of adults with epilepsy," Taylor and Francis, p. 4, 2016.
20 G. R. walters M. L., "Microchem.," vol. 15, p. 231, 1970.
21 Arunima P.R., Rajeev V., Kurra S.B., Mohan R., Nita S., and et al, "Salivary 8 - Hydroxydeoxyguanosine – avaluable indicator for oxidative DNA damage in periodontal disease," The Saudi J Dental Research, Elsevier , p. 2, 2014.
22 Frank G. Whitby, John D. Phillips, Christopher P. Hill, and et al, "Crystal structure of a biliverdin IXalpha reductase enzyme-cofactor complex," Journal of Molecular Biology, p. 1200, 2002.
Received on 12.06.2019 Modified on 18.07.2019
Accepted on 20.08.2019 © RJPT All right reserved
Research J. Pharm. and Tech. 2020; 13(1): 319-322.
DOI: 10.5958/0974-360X.2020.00064.5