European Journal of Cardiovascular Medicine

Iron deficiency anemia is the most public health problem in India. Iron is  most essential element for erythropoiesis,  immunity , cellular immune  system, and oxidative stress (1). WHO determines  hemoglobin <11  g/dL is considered as Anemia  and Severe IDA  (hypochromic, microcytic, Hb <7 g/dL, low serum iron level, and serum ferritin.  IDA is associated with oxidative stress ,  imbalanced condition between reactive oxygen species and antioxidant system. This imbalance can cause irreversible damage to the cellular compartments (2,3). IDA has impact on the development of immunity, cognitive, neurological manifestations (4,5). IDA and secondary thrombocytosis observed in animal models, further they were identified independent risk factors for the development of thrombus  formation and propagation (6).

Antioxidant or pro -oxidants  disparity can cause oxidative  and cellular damage .Further more its imbalance prone to  infectious diseases, diabetes mellitus, cardiovascular, neurodegenerative and cancer pathologies (7,8). Lipid peroxidation is  plays a pivotal role in the pathogenesis of several clinical disorders. The continuous generation of  free radicals on  mono& polyunsaturated fatty acids or phospholipids  in cellular membranes is causative  factor of lipid peroxidation (9,10).  Malondialdehyde is widely accepted  biomarker for the lipid peroxidation (11). There were limited and debatable data with view  oxidative stress and  total antioxidant capacity in iron deficiency anemia. So in this view we would like to evaluate the Malondialdehyde  (MDA)  , total antioxidant capacity levels in IDA patients compared with healthy volunteers, and to find out their association with hemoglobin levels.

tudy Design and Participants

Fifty patients diagnosed with iron deficiency anemia (IDA), belonging to both sexes and various age groups, attending the Government General Hospital attached to Siddhartha Medical College, Vijayawada, Andhra Pradesh, India, were enrolled for this study. An equal number of age- and sex-matched healthy individuals were included as controls.

Selection Procedure

Participants were recruited from the outpatient department after completing a structured questionnaire capturing demographic information such as age, gender, occupation, and geographical location. Detailed medical history, anemia-related clinical profile, and current or past medication use were documented systematically.

Inclusion and Exclusion Criteria

Patients with confirmed IDA willing to provide written informed consent were included. Individuals with acute or chronic infections, chronic systemic illness, other hemoglobinopathies, or those declining consent were excluded.

Biochemical Analysis

The total antioxidant capacity (TAC) of serum samples was estimated using the method described by Benzie et al. [12]. Lipid peroxidation was quantified by measuring malondialdehyde (MDA) levels following the protocol of Mahfouz et al. [13].

Hematological Assessment

Complete hematological parameters, including hemoglobin concentration, red cell indices, and related variables, were measured using an automated hematology analyzer calibrated according to manufacturer specifications.

Sample Handling and Laboratory Procedures

All blood samples were collected under aseptic conditions, transported immediately to the laboratory, and processed within the recommended time frame to preserve biochemical integrity. Standard quality-control measures and reagent validation were observed throughout the study.

Statistical Analysis

Data from hematological indices, antioxidant status, and oxidative stress markers were expressed as mean ± standard deviation. The comparison between controls and iron deficiency anemia (IDA) patients was performed using the Student’s t-test, which revealed significant reductions in hemoglobin, MCV, MCH, PCV, and total antioxidant capacity, along with significantly elevated MDA levels in the IDA group (p < 0.001). Parameters that did not differ significantly, such as WBC, RBC, and platelet counts, were also documented. Correlation analysis was carried out using Pearson’s correlation coefficient (r) to determine the relationship between hemoglobin levels and oxidative stress markers within the study group. A positive correlation was observed between hemoglobin and total antioxidant capacity (r = 0.359, p = 0.04), whereas MDA showed a significant negative correlation with hemoglobin (r = –0.463, p = 0.01). A p-value < 0.05 was considered statistically significant for all analyses.

Ethical Considerations

Written informed consent was obtained from all participants prior to enrollment. The study protocol received approval from the Institutional Ethics Committee of Government Siddhartha Medical College, Vijayawada(IEC/SMC/GGH/2023/AP/144 dated 20 November 2023), and all procedures were conducted in accordance with institutional ethical guidelines.

Table 1: Hematological & antioxidant, oxidative stress parameters in controls and study groups.

*   p value <0.001, # p value <0.05

Table 2: Correlation between Hb  & oxidative stress parameters in study subjects

*Correlation is significant at the 0.05 level (2-tailed).

Iron deficiency is most severe health burden in India and worldwide. Iron is vital element for the synthesis of hemoglobin. The exhaustion of iron stores may result from blood loss, less intake, malabsorption. IDA could be result of occult gastrointestinal bleeding. The Hb levels  IDA  will demonstrate a low mean corpuscular hemoglobin volume (14). In the present study we observed significantly low Hb, MCV , PCV values compared to healthy volunteers. Further deprived ferritin is also one of the reliable indicator  of IDA. But ferritin level that is within the range or elevated is not a significant in patients with other inflammatory conditions such as infection, collagen disease and malignancies (15).

 Reactive oxygen species (ROS) such as O2·-, H2O2, OH·-  and peroxides, nitrosamines  are continuously  generated in all cellular organisms which response to internal and external stimulus.  High concentration of free radicals and low concentration of antioxidants contributing to phagocytic bactericidal activity results oxidative stress lead to metabolic functions (16). In the present study we observed there was significant increased MDA and decreased TAC levels in IDA patients compared with healthy volunteers.

Erythrocytes are remarkabl  handle intracellular oxidative stress through the different  enzymes such as  catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and reduced glutathione (GSH). But deficiency of antioxidants such as Vitamins C & E, carotenoids, and polyphenols may induce  lipid peroxidation and carbonylation of oxidant species . This would predominant causative factor oxidative stress and vascular complications in IDA patients (17,18). Our results suggests that deprived total antioxidant capacity and increased lipid peroxidation in IDA patients and further Hb levels  showed positive correlation with TAC, and negative correlation with MDA . So supplementation of nutrients and balanced diet will beneficial to reduce the risk of  complications in IDA patients.

Decreased Total antioxidant capacity , increased lipid peroxidation  vital factors responsible for increased oxidative stress in IDA patients.  Regular monitoring and supplementation of iron and other multivitamins are beneficial for reduction of oxidative stress and to reduce  iron deficiency  anemia and related complications.

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