Atormac
brintellex
Neurology India
menu-bar5 Open access journal indexed with Index Medicus
  Users online: 5998  
 Home | Login 
About Editorial board Articlesmenu-bullet NSI Publicationsmenu-bullet Search Instructions Online Submission Subscribe Videos Etcetera Contact
  Navigate Here 
 Search
 
  » Next article
  » Previous article 
  » Table of Contents
  
 Resource Links
  »  Similar in PUBMED
 »  Search Pubmed for
 »  Search in Google Scholar for
 »Related articles
  »  Article in PDF (132 KB)
  »  Citation Manager
  »  Access Statistics
  »  Reader Comments
  »  Email Alert *
  »  Add to My List *
* Registration required (free)  


  In this Article
 »  Abstract
 »  Materials and Me...
 »  Results
 »  Discussion
 »  References

 Article Access Statistics
    Viewed16073    
    Printed339    
    Emailed26    
    PDF Downloaded554    
    Comments [Add]    
    Cited by others 34    

Recommend this journal

   
ORIGINAL ARTICLE
Year : 2003  |  Volume : 51  |  Issue : 1  |  Page : 60-62

Free radical toxicity and antioxidants in Parkinson’s disease


Department of Biochemistry, Kasturba Medical College, Manipal-576119

Correspondence Address:
Kasturba Medical College, Manipal-576119
[email protected]

 » Abstract 

Erythrocyte lipid peroxidation, oxidative hemolysis, erythrocyte antioxidant enzymes, viz. superoxide dismutase, glutathione reductase, glutathione peroxidase, catalase and plasma antioxidants, viz. vitamin A, vitamin E, vitamin C and ceruloplasmin have been determined by spectrophotometric methods in 15 patients with Parkinson’s disease (PD) and in 50 controls. Lipid peroxidation, oxidative hemolysis and plasma ceruloplasmin were significantly higher in PD patients as compared to normals. Erythrocyte antioxidants in PD patients were not significantly different from the controls. However, plasma vitamin C in PD patients was significantly lower than the controls. It is concluded that these patients are under oxidative stress which points to a possible involvement of free radicals in PD.

How to cite this article:
Sudha K, Rao A, Rao S, Rao A. Free radical toxicity and antioxidants in Parkinson’s disease . Neurol India 2003;51:60-2


How to cite this URL:
Sudha K, Rao A, Rao S, Rao A. Free radical toxicity and antioxidants in Parkinson’s disease . Neurol India [serial online] 2003 [cited 2021 Jan 19];51:60-2. Available from: https://www.neurologyindia.com/text.asp?2003/51/1/60/1032


Oxygen-derived free radicals have recently been implicated in pathogenesis of various diseases including atherosclerosis, diabetes mellitus, epilepsy, inflammatory diseases and cancer.[1],[2],[3],[4],[5] Lipid peroxidation induced by free radicals is believed to be one of the major causes of cell membrane damage leading to lysis of cell.[6]
The body possesses a complex protective antioxidant system against these potentially toxic products such as vitamin E, vitamin C, vitamin A, glutathione and antioxidant enzymes. These enzymes include glutathione reductase (GR), glutathione peroxidase (GP), superoxide dismutase (SOD) and catalase (CT).[7]
In Parkinson's disease (PD), there is progressive death of substantia nigral cells leading to less availability of dopamine to the striatum which controls movement. Neurons of substantia nigra (SN) may be particularly vulnerable to oxidant stress, because the oxidative metabolism of dopamine has the potential to generate cytotoxic free radicals. Dopamine can be oxidized by either monoamine oxidase or undergo autooxidation to generate hydrogen peroxide (H2O2). H2O2 can damage the neuron directly or indirectly through the formation of hydroxyl radicals in presence of ferrous ions.[8] Neuromelanin present within the SN neurons has the potential to promote site-specific accumulation and reduction of iron thereby potentiating iron-induced lipid peroxidation and consequent cell death.[9],[10] H2O2 is normally detoxified by reduced glutathione (GSH) in the reaction catalyzed by GP, thus an increased rate of dopamine turnover or a deficiency of GSH could lead to oxidative stress. Thus, it appears that free radicals may be one of the important agents responsible for destruction of SN neurons, thereby leading to PD.
However, at present, very few reports[11],[12],[13],[14],[15] are available on antioxidants of blood in patients with PD. Hence, the aim of the study is to evaluate various blood antioxidants and establish the possibility of oxidant damage to the RBC in PD.

  »   Materials and Methods
 Top

The study population consisted of 15 patients who had PD and 50 age and sex matched healthy controls. Diagnosis of PD was mainly by taking a detailed history and studying the symptoms of the disease. The diagnostic criteria for PD were based on those given by Hughes et al.[16] Most of the patients showed bradykinesia and resting tremor. One patient suffered from mild hypertension and another from early dementia. The patients were in the age group of 40-60 years and in the initial stage of the disease (1-2 years) without any drug therapy.
Random blood samples were collected in EDTA bottles from normal subjects and patients. Blood was centrifuged at 3000 g for 10 minutes. Plasma was separated, buffy coat was carefully removed and separated erythrocytes washed thrice with 0.01M saline phosphate buffer pH 7.4 (containing 0.15 M NaCl), then diluted 1:2 with the same buffer and stored at 4-50C. The hemoglobin content of the erythrocytes was determined by cyanmethemoglobin method. Erythrocyte enzymes were estimated in appropriately diluted hemolysates prepared by the addition of distilled water. Erythrocyte GR and GP activity was determined by recording the decrease in absorbance due to depletion of NADPH at 340 nm for a period of 5 minutes.[17],[18] SOD was determined according to the method of Beauchamp and Fridovich[19] based on inhibition of nitroblue tetrazolium reduction. CT activity in the hemolysate was determined by the method of Brannan et al.[20] The assay is based on the disappearance of H2O2 in the presence of the enzyme source at 260C. The lipid peroxidation and oxidative hemolysis of RBC were determined by incubating RBC suspension in saline phosphate buffer containing 0.44M H2O2 at 370C for a period of 2 hours.[21],[22] Aliquots were withdrawn from the above mixture at 0 hour and 2 hours. Lipid peroxidation in RBC was determined by estimating malondialdehyde (MDA) produced using thiobarbituric acid (TBA).
Plasma ceruloplasmin was determined by its p-phenylene diamine oxidase activity.[23] Plasma alpha tocopherol was measured by the Emmorie Engel reaction given by Bieri et al.[24] Vitamin A was determined by reading the extinction at 327 nm before and after exposure to UV light.[25] Plasma vitamin C was determined chemically using dintrophenyl hydrazine as a color compound.[26]
Data was analyzed statistically by Mann Whitney U test. The difference of P < 0.05 was considered significant.

  »   Results Top


The erythrocyte lipid peroxidation in PD patients was significantly high at 0 hour compared to the healthy controls. However after 2 hours of incubation of RBC with H2O2 the increase in lipid peroxide levels of PD patients was not statistically significant. Oxidative hemolysis was significantly high both at 0 hour and 2 hours in PD patients as compared to controls [Table - 1].
A comparison of erythrocyte GR, GP, SOD and CT activities in PD patients to that of controls showed no significant change [Table - 2]. However plasma ceruloplasmin concentration was significantly higher in PD patients than in normal subjects. The mean plasma vitamin C in PD patients was significantly lower compared to that of controls but vitamin A and E levels were within normal range [Table - 3].

  »   Discussion Top


In PD, the environment within SN is conducive to the formation of cytotoxic free radicals. These free radicals react instantaneously with membrane lipids and cause lipid peroxidation and cell death.[27] In the present study, there is significantly high erythrocyte lipid peroxidation at 0 hour which supports the notion of increased lipid peroxidation in the PD brain as well.[28] Lipid peroxidation of RBC membrane causes them to lose their ability to change shape and squeeze through the smallest capillaries, thus eventually leading to hemolysis. Jenner et al[28] have shown an enhanced basal lipid peroxidation in SN of post mortem brain in PD. In MPTP (a neurotoxin producing PD) treated rats the basal lipid peroxide concentration is significantly high in SN.[29] Brain tissue extracts of PD patients showed a tenfold increase in lipid hydroperoxides in SN compared to control subjects.[30] This evidence indicates that increased lipid peroxidation could possibly play a role in neurodegeneration leading to PD. The present study also shows similar changes in blood where increased lipid peroxidation has lead to lysis of erythrocytes. This finding is also supported by the work on lipid peroxides in RBC elsewhere.[15]
Moreover, in this study, there has been no change observed in the antioxidant enzyme levels of RBC in PD. Though the mean values of these enzymes were lower in PD patients compared to normal, none of them were statistically significant. This observation is supported by the reports made on post mortem brains and in MPTP treated animals, where activities of CT, GP and GSH concentrations in SN remained unchanged compared to controls.[28],[29] Further, several studies indicated a decreased SOD activity in blood of PD patients.[12],[13],[14] In contrast, increase in erythrocyte SOD level in PD has also been reported.[15]
Significantly high plasma ceruloplasmin level in PD patients observed in the present study may be attributed to its ferroxidase activity. The ferroxidase activity inhibits iron dependent lipid peroxidation[31] and also formation of hydroxyl radicals. Good et al[32] have demonstrated that iron-induced oxidant stress contributes to cell death in PD. However, significantly decreased serum ceruloplasmin levels have also been observed in PD.[14] A significant decrease in plasma vitamin C is seen in PD patients compared to controls in this study. A similar observation is made by Youdim et al[10] who have shown low vitamin C levels in SN of PD patients. Addition of vitamin C to an incubation mixture containing tissue from different brain areas and ferrous sulfate, decreased lipid peroxidation in SN of MPTP treated monkeys.[29] Low plasma vitamin C level in PD patients as observed here is probably due to increased utilization of the vitamin to mitigate the toxicity of free radicals.
On the whole, it can be concluded that erythrocytes of PD patients are under oxidative stress as evidenced by increased lipid peroxidation and oxidative hemolysis. These findings are in keeping with the possible role of free radical damage in PD. 

 » References Top

1.Plachta H, Bartnikowska E, Obara A. Lipid peroxides in blood from patients with atherosclerosis of coronary and peripheral arteries. Clin Chim Acta 1992;211:101-2.  Back to cited text no. 1    
2.Oberley LW. Free radicals and diabetes. Free Radical Biol Med 1988;5:113-24.  Back to cited text no. 2  [PUBMED]  
3.Sudha K, Rao AV, Rao A. Oxidative stress and antioxidants in epilepsy. Clin Chim Acta 2001;303:19-24.  Back to cited text no. 3  [PUBMED]  [FULLTEXT]
4.Fantone JC, Ward PA. Role of oxygen derived free radicals and metabolites in leukocyte-dependent inflammatory reactions. Am J Pathol 1982;107:397-418.  Back to cited text no. 4    
5.Sundstrom H, Korpela H, Viinikka L, Kauppila A. Serum selenium and GP and plasma lipid peroxides in uterine, ovarian or vulvar cancer, and their responses to antioxidant in patients with ovarian cancer. Cancer Lett 1984;24:1-10.   Back to cited text no. 5  [PUBMED]  
6.Barber AA, Bernheim F. Lipid peroxidation its measurement occurrence and significance in animal tissues. Adv Gerontol Res 1967;2:355-403.  Back to cited text no. 6  [PUBMED]  
7.Halliwell B, Gutteridge JMC. The antioxidants of human extracellular fluids. Arch Biochem Biophys 1990;280:1-8.  Back to cited text no. 7    
8.Cohen G. Oxygen radicals and Parkinson's disease. In: Proceedings of Upjohn Symposium, Fed Amer Soc Exp Biol (Bethesda, MD) 1987. pp. 130-5.  Back to cited text no. 8    
9.Youdim MB, Ben-Shachar D, Riederer P. The possible role of iron in the etiopathology of Parkinson's disease. Mov Disord 1993;8:1-12.  Back to cited text no. 9  [PUBMED]  
10.Youdim MB, Riederer P. Understanding Parkinson's disease. Sci Am 1997;1:38-45.  Back to cited text no. 10    
11.Munch G, Gerlach M, Sian J, Wong A, Riederer P. Advanced glycation end products in neurodegeneration: more than early markers of oxidative stress? Ann Neurol 1998;44:S85-8.  Back to cited text no. 11  [PUBMED]  
12.Bostantjopoulou S, Kyriazis G, Katsarou Z, Kiosseoglou G, Kazis A, Mentenopoulos G. Superoxide dismutase activity in early and advanced Parkinson's disease. Funct Neurol 1997;12:63-8.  Back to cited text no. 12  [PUBMED]  
13.Ihara Y, Chuda M, Kuroda S, Hayabara T. Hydroxyl radical and superoxide dismutase in blood of patients with Parkinson's disease: Relationship to clinical data. J Neurol Sci 1999;170:90-5.  Back to cited text no. 13  [PUBMED]  [FULLTEXT]
14.Torsdottir G, Kristinsson J, Sveinbjornsdottir S, Snaedal J, Johanhesson T. Copper, ceruloplasmin, superoxide dismutase and iron parameters in Parkinson's disease. Pharmacol Toxicol 1999;85:239-43.  Back to cited text no. 14    
15.Serra JA, Dominguez RO, de Lustig ES, Guareschi EM, Famulari AL, Bartolome EL, et al. Parkinson's disease is associated with oxidative stress: Comparison of peripheral antioxidant profiles in living Parkinson's Alzheimer's and vascular dementia patients. J Neurol Transm 2001;108:1135-48.  Back to cited text no. 15  [PUBMED]  [FULLTEXT]
16.Hughes AJ, Daniel SE, Blankson S, et al. A clinicopathologic study of 100 cases of Parkinson's disease. Arch Neurol 1993;50:140-8.  Back to cited text no. 16  [PUBMED]  
17.Horn HD, Burns FH. In: Bergmeyer HV, editor. Methods of Enzymatic Analysis. New York: Academic Press; 1978. pp. 875.  Back to cited text no. 17    
18.Paglia DE, Valentine WN. Studies on the quantitative and qualitative characterization of erythrocyte GP. J Lab Clin Med 1967;70:158-69.  Back to cited text no. 18  [PUBMED]  
19.Beauchamp C, Fridovich I. Superoxide dismutase: improved assays and an assay applicable to acrylamide gels. Anal Biochem 1971;44:276-87.  Back to cited text no. 19  [PUBMED]  
20.Brannan TS, Maker HS, Raes IP. Regional distribution of CT in the adult rat brain. J Neurochem 1981;6:307-9.  Back to cited text no. 20    
21.Stocks J, Dormanday TL. The autooxidation of human red cell lipids induced by hydrogen peroxide. Br J Haematol 1971;20:95-111.  Back to cited text no. 21    
22.Kartha VN, Krishnamurthy S. Effect of hyper vitaminosis A on hemolysis and lipid peroxidation in the rat. J Lipid Res 1978;19:332-34.  Back to cited text no. 22    
23.William Sunderman F, Jr Shozo Nomoto. Measurement of human serum ceruloplasmin by its p-phenylene diamine oxidase activity. Clin Chem 1970;16:  Back to cited text no. 23    
24.903-10.  Back to cited text no. 24    
25.Bieri JG, Teets L, Belavady B, Andres EL. Serum vitamin E levels in a normal adult population in the Washington, D.C. area. Proc Soc Exptl Bio Med 1964;117:131-3.  Back to cited text no. 25    
26.Paterson JCS, Wiggins HS. An estimation of plasma vitamin A and the vitamin A absorption test. J Clin Path 1954;7:56-60.  Back to cited text no. 26    
27.Omaye ST, Turnbull JD, Sauberlich HE. Selected methods for the determination of ascorbic acid in animal cells, tissues and fluids. Methods Enzymol 1979;62:3-11.  Back to cited text no. 27  [PUBMED]  
28.Menon NK, Furdik JV, Mishra K. Role of oxygen free radicals in neurological disorders. Advances in clinical Neurosciences 1994;4:327-39.  Back to cited text no. 28    
29.Jenner P, Dexter DT, Sian J, Schapira AH, Marsden CD. Oxidative stress as a cause of nigral cell death in Parkinson's disease and incidental Lewy body disease. The royal kings and queens Parkinson's disease research group. Ann Neurol 1992;32:S82-7.  Back to cited text no. 29  [PUBMED]  
30.Marzatico F, Cafec, Taborelli M, Benzi G. Experimental Parkinson's disease in monkeys. Effect of ergot alkaloid derivative on lipid peroxidation in different brain areas. Neurochem Res 1993;18:1101-6.  Back to cited text no. 30    
31.Dexter DT, Holley AE, Flitter WD, Slater TF, Wells FR, Daniel SE, et al. Increased levels of lipid hydroperoxides in the Parkinsonian substantia nigra: an HPLC and ESR study. Mov Disord 1994;9:92-7.  Back to cited text no. 31  [PUBMED]  
32.Gutteridge JMC, Richamond R, Halliwell B. Oxygen free radicals and lipid peroxidation: inhibition by the protein ceruloplasmin. FEBS Lett 1980;112:269.  Back to cited text no. 32    
33.Good PF, Olanow CW, Perl DP. Neuromelanin containing neurons of the substantia nigra accumulate iron and aluminium in Parkinson's disease: a LAMMA study. Brain Res 1992;593:343-6.  Back to cited text no. 33  [PUBMED]  

 

Top
Print this article  Email this article
Previous article Next article
Online since 20th March '04
Published by Wolters Kluwer - Medknow