Home About us Editorial board Search Ahead of print Current issue Archives Submit article Instructions Subscribe Contacts Login 
Visit old site
Home Print this page Email this page Small font size Default font size Increase font size
Users Online: 485


 
 Table of Contents  
ORIGINAL ARTICLE
Year : 2015  |  Volume : 7  |  Issue : 5  |  Page : 194-198

Status of Superoxide dismutase in transfusion dependent thalassaemia


1 Department of Molecular Biology, Medical Faculty of Jenderal Soedirman University, Purwokerto, Indonesia
2 Department of Pediatric, Sardjito National Hospital, Yogyakarta, Indonesia
3 Department of Biochemistry, Medical Faculty of Yayasan Rumah Sakit Islam Indonesia University, Jakarta, Indonesia

Date of Web Publication26-May-2015

Correspondence Address:
Lantip Rujito
Medical Faculty of Jenderal Soedirman University, Purwokerto
Indonesia
Login to access the Email id

Source of Support: None, Conflict of Interest: None


DOI: 10.4103/1947-2714.157480

Rights and Permissions
  Abstract 

Background: Thalassemia is a collection of genetic impairments in beta and alpha genes causing various states of anemia. Severe types of the disease need lifelong transfusions, leading to oxidant-antioxidant disturbance due to massive iron deposits. Aims: The aim of this study was to assess the antioxidant enzyme Superoxide Dismutase (SOD) and ferritin levels of thalassemia major patients in a peripheral health facility. Materials and Methods: Two hundred and nine probands were recruited and performed laboratory experiments for SOD and Ferritin levels. Chelation administration and clinical score were taken from interviewing the family and from medical report data. Results: The study showed that SOD intensity was lower (162.41 u/ml) compared to the normal cutoff point (P = 0.001), while the mean of Ferritin levels was ten times over the normal value (4226,67 ng/dl). Observations also reported that chelation medicine was not administrated properly. Conclusions: The data indicates that thalassemic patients have oxidant-antioxidant uproar due to oxidative stress. Monitored chelating administration, selective antioxidant, and a well-balanced diet may prevent oxidative injury.

Keywords: Oxidant-antioxidant, superoxide dismutase, thalassaemia


How to cite this article:
Rujito L, Mulatsih S, Sofro AM. Status of Superoxide dismutase in transfusion dependent thalassaemia. North Am J Med Sci 2015;7:194-8

How to cite this URL:
Rujito L, Mulatsih S, Sofro AM. Status of Superoxide dismutase in transfusion dependent thalassaemia. North Am J Med Sci [serial online] 2015 [cited 2018 Nov 15];7:194-8. Available from: http://www.najms.org/text.asp?2015/7/5/194/157480


  Introduction Top


Thalassemia is a clinical hematology problem caused by a collection of genetic abnormalities in the gene cluster-forming β and α chains of proteins carrying oxygen, hemoglobin. [1] Patients with severe cases cannot produce normal hemoglobin, leading to a life-long anemic state. Treatments for the condition depend on continuous blood transfusions to maintain a good quality of life. However, empirical data shows that regular transfusions lead to an iron overload with a massive increase in non-transferrin-bound iron that may cause more tissue damage than conjugated iron. Further results of these processes are the occurrences of excessive oxidative stress and disturbance defense of oxidant-antioxidant mechanisms. [2]

Iron-induced oxidative stress is one of the most important factors determining cell injury in thalassemic patients. It has been reported that reactive oxygen species (ROS) involved in red cells are damaged due to increased membrane rigidity, deformity, and induced hemolysis. [3] In addition, oxidative stress has been recognized as initiating the removal of red cells by the immune system. [4] Endogenous antioxidants, like superoxide dismutase (SOD), Catalase and glutathione peroxidase (GPx) are the first barriers to the change of the internal environment influenced by the increase of free radicals and abundant stress, creating superactiveoxygen. However, much of the data from thalasemic patients state that SOD level can vary from a low level, no different from healthy individuals, up to a high level. [5],[6],[7] We assume that the variation may be caused by different areas and management. The purpose of this study was to assess the status of the SOD enzyme in transfusion-dependent thalassemic subjects in Banyumas, a remote region in Central Java, Indonesia.


  Materials and Methods Top


Research Subjects

Subjects were patients with Thalassaemia major that were diagnosed based on clinical symptoms, blood analysis index, and hemoglobin electrophoresis. Two hundred and nine subjects aged 6 months to 65 years were included in this study, excluding patients with hypothyroidism and hyperthyroidism, diabetes mellitus, and acute inflammation. Subjects read and signed informed consent waivers approved by the Medical Faculty Ethic Committee.

Laboratory Experiments

Ten milliliters of blood were drawn and stored in EDTA tubes to be used in laboratory experiments. Levels of (SOD) activity in red blood cells were assayed by RANSOD Kit (Randox, United Kingdom) and were expressed in U/ml. Excess of ferritin was measured using ELISA Kit for ferritin (Sigma-Aldrich, USA) and counted in ng/dl. To determine clinical scores of thalasemic patients, the Sripichai Score was adopted. [8]

Statistical Analysis

Data was presented in a descriptive manner, including mean and standard deviation. Comparison between the case and normal cut-off point was performed using one sample t test. Person's correlation was used to determine the relationship between SOD and clinical appearances.


  Results Top


During the study period, ferritin value was collected 3-6 times, reflected in serial retrieval. The mean of serum ferritin among subjectswas 4226,67 ng/dl, significantly increased from standard normal value, for both children and adults. This study also reported that the average level of SOD activity was 162.41 u/ml. Compared with the normal population, it was below the standard value (the average varying from 164-240 u/ml) [Table 1] and [Figure 1]. Approximately sixty percent of the patients in our study had serum ferritin exceeding 2500 ng/dl, which is almost ten times higher than the upper limit of normal. Ferritin level of the patients was depicted in [Table 2]. [Figure 1] depicts that in general, red cell SOD activity in thalasemic patients studied were below average, however some individuals expressed high SOD activity. One sample t-test performed on the mean of SOD activity expressed lower value than the mean of normal individuals (P = 0.001). Using Person's corelation statistic, no relationship resulted between SOD and degree of clinical patients (P = 0.66).
Figure 1: SOD Distribution among reaseach subjects. It show that SOD level among thalassaemia patients are under normal value (164-264 u/ml)

Click here to view
Table 1: The mean of ferritin and SOD and their correlation with clinical score

Click here to view
Table 2: Ferritin levels according to the mean age of patients

Click here to view



  Discussion Top


Ferritin distribution was directly proportional to the age of the patients, as depicted in [Table 2]. Lowest levels were in the age group of <2 years old, followed by 2-10 years. As transfusions became regular treatment, the amountof ferritin increased as manifest of iron from red blood cells accumulation. For subjects >35 years old, low ferritin levels may relate to the onset of transfusion, as they carry a mild form of Thalassaemia. Patients with moderate severity, regardless of genotype, were characterized usually by moderate anemia and required no or only infrequent red blood cell transfusions. [9]

It was already known that ferritin from thalassemia patients could reach >12,000 ng/dl, and the iron chelating program must be a priority when the threshold (1000 ng/dl) has been exceeded. [10] The program has become part of the management of thalassaemiain local hospitals by means of the medicine, deferiprone. However, the study indicates that patient adherence to administrate chelation was not monitored well. It could be characterized by looking at high ferritin levels in the study subjects. Previous data showed that deferiprone was effective only in the initial conditions of high ferritin (>5000 ng/dl), whereas in 2500-5000 ng/dl levels, ferritin tends to be stagnate. [11] Later studies also displayed similar conclusions; decreasing ferritin with deferiprone was significantly less likely to succeed with lower initial values. [12]

The study also found a correlation between ferritin and clinical score appearance. The score indicates that the degree of clinical impairment is in line with higher ferritin levels in patients (P = 0.04). As in previous studies, the data reaffirmed that the excessive iron deposits will affect the clinical development, mainly growth and co-morbid complicating factors. [13] Iron deposits in metabolicorgans (such as the liver, pancreas, and spleen) may develop severe complications caused by disturbance mechanisms, including the immune system, oxidant-antioxidant regulation, and metabolism disruption.

Despite the facts, plasma ferritin is also influenced by the state of acute infection, iron metabolism disorders, and acute physical trauma. [14],[15] Such condition may increase the levels 2 to 30-fold. [16] In other words, other assumptions about the increase of ferritin aside from over-transfusion cannot be ruled out.

Together with GPx, SOD is an intracellular enzyme that is responsible for changes in the oxidant-antioxidant balance in cells. Enzyme function is to catalyze modifying ion free radicals, especially 0 2 - into H2 0. [17] In subjects with thalassaemia, enormous free radicals built up due to the state of iron overload (resulting from transfusions and ineffective erythropoiesis). Iron (Fe) is able to accelerate the change of molecular oxygen into reactive oxygen radicals, superoxide, and hydroxyl groups through the Fenton reaction. [18],[19]

These low levels of SOD activity were in agreement withpreviously published data. [20] Thisstudy showed that patients with homozygous thalassaemia decreased 1.5 times lower than the normal individual. In line with the studies, Patne et al., in 2012 also present data showing that the levels of erythrocyte antioxidant enzymes, especially SOD and GPx activity, decreased significantly in patients who were transfusion-dependent. [21] Another study also concluded that the degree of pain and clinical appearance correlate with low levels of antioxidants. [7] All of these studies suggestthat monthly transfusion leads to decreased SOD and GPx levels. However, different results were shown by other research centers. Simsek and colleagues found that the levels of SOD and GPx in thalassemic patients were higher than the healthy controls and careers, while vitamin E levels were lower. [22] Other publications mention that SOD did not show significant differences between healthy controls and thalassemic subjects. [2]

Increased levels of antioxidants, including SOD, occur in various circumstances: Including an acute inflammatory phase, a state of trauma, and upon exposure to increased levels of pro-oxidants. The increase was associated with a compensatory mechanism to break down free radicals that had been caused by oxidative stress and lipid peroxidation. [23] In achronic clinical state, decline was associated with the inability of the antioxidant system to compensate excessive originators. Free radicals could not be offset by the system, which may have caused the degradation of proteins (including enzymes) and cell membranes, which in turn decreased the levels and activity of antioxidant enzymes. [24] This is supported by other publications, which state that chronic stress in diabetes mellitus, metabolic syndrome, chronic liver disease, SLE, and rheumatoid arthritis affect the decrease in antioxidant enzyme capacity. [25],[26],[27]

In addition, the study of iron overload diseases, such as Hemochromatosis, also found that the total antioxidant capacity will decline. [28] Rat models with Hemochromatosis (HFE gene mutations) showed to have increased iron levels but decreased levels of antioxidant enzymes and non-enzymes. [29] Other findings also stated that Hemochromatosis subjects expressed low levels of the paraoxonase enzyme (one of the peroxidase lipids degradation enzymes) compared to normal individuals. [30]

Variations in findings on the thalassemic subjects may be caused by other factors or mechanisms that play a role, including iron chelation and daily diet of the patients. Research in Jakarta said that the decrease of antioxidant enzymes in patients related to non-chelating subjects [5] , while administration of curcuma on a regular basis, could increase the capacity of the enzymes SOD, GPx, and GSH. The discontinuation, however, caused levels of the enzymesto go back to original value. [31] Iron chelation, including Deferoxamin (DFO) and EDTA, will first bind Fe 2+ , oxidizing reactive Fe 2+ into Fe 3+ , which is more stable. This metal oxidation process significantly lowered oxygen oxidation to become reactive oxygen. [3] In many cases with impaired oxidant-antioxidant mechanisms, administrating an iron chelatorwill improve the prognosis of various disorders, including neurodegenerative disease [32] , cardiovascular impairment [33] , and iron overload. [34],[35] Thus, the presence of varying levels of SOD may be interfered with by the effect of iron chelation management and diet, although other inflammatory factors cannot be ruled out.


  Acknowledgement Top


We give thanks to the provider of the doctoral grant, the Ministry of National Education, Indonesia. Thanks also goes to Dr. Yayan for laboratory preparation.

 
  References Top

1.
Weatherall DJ. Thalassemia as a global health problem: Recent progress toward its control in the developing countries. Ann N Y Acad Sci 2010;1202:17-23.  Back to cited text no. 1
    
2.
Ghone RA, Kumbar KM, Suryakar AN, Katkam RV, Joshi NG. Oxidative stress and disturbance in antioxidant balance in beta thalassemia major. Indian J Clin Biochem 2008;23:337-40.  Back to cited text no. 2
    
3.
Jomova K, Valko M. Importance of iron chelation in free radical-induced oxidative stress and human disease. Curr Pharm Des 2011;17:3460-73.  Back to cited text no. 3
    
4.
Fulda S, Gorman AM, Hori O, Samali A. Cellular stress responses: Cell survival and cell death. Int J Cell Biol 2010;2010:214074.  Back to cited text no. 4
    
5.
Laksmitawati DR, Handayani S, Udyaningsih-Freisleben SK, Kurniati V, Adhiyanto C, Hidayat J, et al. Iron status and oxidative stress in beta-thalassemia patients in jakarta. Biofactors 2003;19:53-62.  Back to cited text no. 5
    
6.
Abdalla MY, Fawzi M, Al-Maloul SR, El-Banna N, Tayyem RF, Ahmad IM. Increased oxidative stress and iron overload in jordanian β-thalassemic children. Hemoglobin 2011;35:67-79.  Back to cited text no. 6
    
7.
Kattamis C, Lazaropoulou C, Delaporta P, Apostolakou F, Kattamis A, Papassotiriou I. Disturbances of biomarkers of iron and oxidant-antioxidant homeostasis in patients with beta-thalassemia intermedia. Pediatr Endocrinol Rev 2011;8:256-62.  Back to cited text no. 7
    
8.
Sripichai O, Makarasara W, Munkongdee T, Kumkhaek C, Nuchprayoon I, Chuansumrit A, et al. A scoring system for the classification of beta-thalassemia/Hb E disease severity. Am J Hematol 2008;83:482-4.  Back to cited text no. 8
    
9.
Ginzburg Y, Rivella S. Beta-thalassemia: A model for elucidating the dynamic regulation of ineffective erythropoiesis and iron metabolism. Blood 2011;118:4321-30.  Back to cited text no. 9
    
10.
Galanello R, Origa R. Beta-thalassemia. Orphanet J Rare Dis 2010;5:11.  Back to cited text no. 10
    
11.
Cohen. AR, Galanello R, Piga A, Dipalma A, Vullo C, Tricta F. Safety profile of the oral iron chelator deferiprone: A multicentre study. Br J Haematol 2000;108:305-12.  Back to cited text no. 11
    
12.
Cohen AR, Galanello R, Piga A, De Sanctis V, Tricta F. Safety and effectiveness of long-term therapy with the oral iron chelator deferiprone. Blood 2003;102:1583-7.  Back to cited text no. 12
    
13.
Xu, LH, Fang JP, Xu HG, Weng WJ. Evaluation of hepatic iron overload in Chinese children with beta-thalassemia major. Pediatr Hematol Oncol 2011;28:702-7.  Back to cited text no. 13
    
14.
Ruslianti V, Chairulfatah A, Rachmadi D. Hubungan spektrum klinis infeksi dengue dengan kadar seng dan feritin serum. Sari Pediatr 2013;15:213-9.  Back to cited text no. 14
    
15.
Peeling P, Sim E, Badenhorst CE, Dawson B, Govus AD, Abbiss CR, et al. Iron Status and the acute post-exercise hepcidin response in athletes. PLoS One 2014;9:e93002.  Back to cited text no. 15
    
16.
Northrop-Clewes CA. Interpreting indicators of iron status during an acute phase response-lessons from malaria and human immunodeficiency virus. Ann Clin Biochem 2008;45:18-32.  Back to cited text no. 16
    
17.
Fukai T, Ushio-Fukai M. Superoxide dismutases: Role in redox signaling, vascular function, and diseases. Antioxid Redox Signal 2011;15:1583-606.  Back to cited text no. 17
    
18.
Winterbourn CC. Toxicity of iron and hydrogen peroxide: The Fenton reaction. Toxicol Lett 1995;82-83:969-74.  Back to cited text no. 18
    
19.
Shazia, Q, Mohammad ZH, Rahman T, Shekhar HU. Correlation of oxidative stress with serum trace element levels and antioxidant enzyme status in beta thalassemia major patients: A review of the literature. Anemia 2012;2012:270923.  Back to cited text no. 19
    
20.
Dhawan V, Kumar KhR, Marwaha RK, Ganguly NK. Antioxidant status in children with homozygous thalassemia. Indian Pediatr 2005;42:1141-5.  Back to cited text no. 20
    
21.
Patne AB, Hisalkar PJ, Gaikwad SB, Patil SV. Alterations in antioxidant enzyme status with lipid peroxidation in thalassemia major patients. Int J Pharm Life Sci 2012;3:2003-6.  Back to cited text no. 21
    
22.
Simsek F, Öztürk G, Kemahlý S, Erbaþ D, Hasanoðlu A. Oxidant and antioxidant status in beta thalassemia major patients. Ankara üniversitesi Týp Fakültesi Mecmuasý 2005;58:34-8.  Back to cited text no. 22
    
23.
Lü JM, Lin PH, Yao Q, Chen C. Chemical and molecular mechanisms of antioxidants: Experimental approaches and model systems. J Cell Mol Med 2010;14:840-60.  Back to cited text no. 23
    
24.
Willcox JK, Ash SL, Catignani GL. Antioxidants and prevention of chronic disease. Crit Rev Food Sci Nutr 2004;44:275-95.  Back to cited text no. 24
    
25.
Ramakrishna V, Jailkhani R. Evaluation of oxidative stress in Insulin Dependent Diabetes Mellitus (IDDM) patients. Diagn Pathol 2007;2:22.  Back to cited text no. 25
    
26.
Cardin R, Piciocchi M, Bortolami M, Kotsafti A, Barzon L, Lavezzo E, et al. Oxidative damage in the progression of chronic liver disease to hepatocellular carcinoma: An intricate pathway. World J Gastroenterol 2014;20:3078-86.  Back to cited text no. 26
    
27.
Hassan SZ, Gheita TA, Kenawy SA, Fahim AT, El-Sorougy IM, Abdou MS. Oxidative stress in systemic lupus erythematosus and rheumatoid arthritis patients: Relationship to disease manifestations and activity. Int J Rheum Dis 2011;14:325-31.  Back to cited text no. 27
    
28.
Young IS, Trouton TG, Torney JJ, McMaster D, Callender ME, Trimble ER. Antioxidant status and lipid peroxidation in hereditary haemochromatosis. Free Radic Biol Med 1994;16:393-7.  Back to cited text no. 28
    
29.
Turoczi T, Jun L, Cordis G, Morris JE, Maulik N, Stevens RG, et al. HFE mutation and dietary iron content interact to increase ischemia/reperfusion injury of the heart in mice. Circ Res 2003;92:1240-6.  Back to cited text no. 29
    
30.
Martinelli N, García-Heredia A, Roca H, Aranda N, Arija V, Mackness B, et al. Paraoxonase-1 status in patients with hereditary hemochromatosis. J Lipid Res 2013;54:1484-92.  Back to cited text no. 30
    
31.
Kalpravidh RW, Siritanaratkul N, Insain P, Charoensakdi R, Panichkul N, Hatairaktham S, et al. Improvement in oxidative stress and antioxidant parameters in β-thalassemia/Hb E patients treated with curcuminoids. Clin Biochem 2010;43:424-9.  Back to cited text no. 31
    
32.
Youdim MB, Fridkin M, Zheng H. Novel bifunctional drugs targeting monoamine oxidase inhibition and iron chelation as an approach to neuroprotection in Parkinson's disease and other neurodegenerative diseases. J Neural Transm 2004;111:1455-71.  Back to cited text no. 32
    
33.
Korkmaz S, Barnucz E, Loganathan S, Li S, Radovits T, Hegedus P, et al. Q50, an iron-chelating and zinc-complexing agent, improves cardiac function in rat models of ischemia/reperfusion-induced myocardial injury. Circ J 2013; 77:1817-26.  Back to cited text no. 33
    
34.
Ko BS, Chang CS, Chang MC, Chen TY, Chiou TJ, Chiu CF, et al. Guidelines for treating iron overload in myelodysplastic syndromes: A Taiwan consensus statement. Int J Hematol 2014;100:7-15.  Back to cited text no. 34
    
35.
Shenoy N, Vallumsetla N, Rachmilewitz E, Verma A, Ginzburg Y. Impact of iron overload and potential benefit from iron chelation in low-risk myelodysplastic syndrome. Blood 2014;124:873-81.  Back to cited text no. 35
    


    Figures

  [Figure 1]
 
 
    Tables

  [Table 1], [Table 2]


This article has been cited by
1 Electrogenerated chemiluminescence of lucigenin at mesoporous platinum electrode and its biosensing application to superoxide dismutase
Sungju Nam,Won-Yong Lee
Journal of Electroanalytical Chemistry. 2018; 808: 59
[Pubmed] | [DOI]
2 The relationship between ferritin levels and oxidative stress parameters in serum of ß-thalassemia major patients
Ziya Salman,Tamer Yilmaz,Güldal Mehmetçik
Archives of Biochemistry and Biophysics. 2018; 659: 42
[Pubmed] | [DOI]
3 Label-free electrochemical immunosensor for the rapid and sensitive detection of the oxidative stress marker superoxide dismutase 1 at the point-of-care
Paulraj Santharaman,Mainak Das,Sushil K. Singh,Niroj K. Sethy,Kalpana Bhargava,Jonathan C. Claussen,Chandran Karunakaran
Sensors and Actuators B: Chemical. 2016; 236: 546
[Pubmed] | [DOI]



 

Top
 
 
  Search
 
Similar in PUBMED
   Search Pubmed for
   Search in Google Scholar for
 Related articles
Access Statistics
Email Alert *
Add to My List *
* Registration required (free)

 
  In this article
Abstract
Introduction
Materials and Me...
Results
Discussion
Acknowledgement
References
Article Figures
Article Tables

 Article Access Statistics
    Viewed1685    
    Printed21    
    Emailed1    
    PDF Downloaded226    
    Comments [Add]    
    Cited by others 3    

Recommend this journal