Relation between levels of transforming growth factor beta 1 and ferritin in β-thalassemic patients

Abstract
Beta-thalassemia is a recessive hereditary genetic disease characterized by a mutation in the beta-globin gene. Recent studies divide beta-thalassemia patients into transfusion-dependent thalassemia (TDT) and nontransfusion-dependent thalassemia (NTDT) according to their transfusion requirements which makes TDT a major clinical condition and big health problem and community burden especially in developing countries. The hallmark in the pathogenesis of thalassemia is iron overload, which is responsible for most of the complications and end organ damage. Iron status can be assessed by serum ferritin, which is iron-binding protein. Transforming growth factor beta 1 (TGF-β1) is one of the polypeptide members of the cytokines superfamily, which plays a wide range of multicellular functions. Here in our study, our aim was to assess the relationship between TGF-β1 and serum ferritin in the Egyptian population.

Context
This study was conducted to find a relation between levels of TGF-β1 and ferritin in β-thalassemic patients.


Aims
Eighty thalassemic patients were selected from the Medical Research Institute and Alexandria Main University Hospitals, which then were grouped into two groups: group 1 TDT and group 2 NTDT, together with 80 controls of matched age and sex. We aimed to study if there is a relation between levels of TGF-β1 and ferritin in β-thalassemic patients. And also correlate levels of TGF-β1 and ferritin between TDT and NTDT.


Settings and design
Case–control study.


Patients and methods
We obtained written consent from patients and controls. The study had the approval of the Alexandria Medical Ethics Committee. Full history was taken from all patients, including demographic data, clinical data (splenomegaly or history of splenectomy) retrieved from patients, and their transfusion history. Ten centimeters of venous blood were taken from patients prior to transfusion to measure TGF-β1 and serum ferritin. All reagents were brought to laboratory at room temperature for 20 min before use. Put unused solution back at 2–8°C. Dilute the SABC with SABC dilution buffer at 1: 100 and mix them thoroughly (i.e. add 1 μl of SABC into 99 μl of SABC dilution buffer). Assay procedure step 1: wash plate two times before adding standard, sample (diluted at least 1/2 with sample dilution buffer), and control (blank) wells! Step 2: add 100 μl standard or sample to each well and incubate for 90 min at 37°C. Wash step: aspirate and wash plates two times. Step 3: add 100 μl biotin-labeled antibody working solution to each well and incubate for 60 min at 37°C. Wash step: aspirate and wash plates three times. Step 4: add 100 μl SABC working solution into each well and incubate for 30 min at 37°C. Wash step: aspirate and wash plates five times. Step 5: add 90 μl TMB substrate solution. Incubate 10–20 min at 37°C. Step 6: add 50 μl stop solution. Read at 450 nm immediately and calculate.


Statistical analysis used
The data was collected and entered into the personal computer. Statistical analysis was done using Statistical Package for Social Sciences (SPSS/version 24) software.
Quantitative data were described using mean and SD for normally distributed data, while abnormally distributed data was expressed using median, minimum, and maximum. For numerical data, a t test was used to compare two groups, while for more than two groups analysis of variance test was used. For nonparametric data, Kruskal–Wallis test was used. To find the association between two variables, the Spearman correlation coefficient test was used. The level of significance was 0.05.


Results
In a comparison between cases and control regarding levels of TGF-β1 and ferritin in β-thalassemic patients, a significant relationship was confirmed. On the other hand, no relation between two parameters was found in comparison between two groups (TDT and NTDT).


Conclusions
Higher TGF-β1 levels in thalassemics might be an extraordinary cause of disturbing iron metabolism, expressed by elevated serum ferritin levels.