Determination of humanin and MOTS-C peptide levels in thalassemia patients and evaluation of their relationship with nitrosative stress and apoptosis

Objective: Thalassemia is an autosomal recessive hypochromic microcytic anemia and is a heterogeneous group of diseases characterized by defective or insufficient globin chain synthesis in the hemoglobin molecule. This is a group of patients who are on a regular erythrocyte transfusion program. In thalassemia patients, iron accumulation occurs as a result of erythrocyte transfusion. Hemosiderosis is an undesirable and fatal consequence of prolonged transfusion therapy and the iron load cannot be removed from the body physiologically. The purpose of iron chelation therapy is to prevent iron accumulation in the body, reduce existing iron accumulation, and thus prevent complications due to increased body iron accumulation. Iron ions in the environment accelerate lipid peroxidation. Lipid radicals transform into highly cytotoxic products. Reactive oxygen particles formed in the body as a result of iron accumulation cause damage to the endothelium. Damage to the endothelium triggers apoptosis at the cellular level. Our study aimed to show the damage caused by the iron load accumulated in the body in thalassemia patients receiving regular transfusion. Nitrosative stress and apoptosis observed in the body due to this iron load were evaluated and the relationship between Humanin and MOTS-c peptides, which are mitochondrial derived peptides, was evaluated. Materials and Methods: In our study, patients who were followed up with a diagnosis of thalassemia and were transfused with erythrocytes at our Gaziantep University Şahinbey Research and Practice Hospital Faculty of Medicine Pediatric Hematology and Oncology outpatient clinic between October 2022 and December 2023 were divided into 2 groups, those who received chelators and those who did not, and were studied prospectively with a control group. Among the erythrocyte transfused patients diagnosed with thalassemia between the ages of 0-18, the first group (n=34) received iron chelator treatment, the second group (n=21) did not receive iron chelator treatment, and the third group (n=28) was the control group; It was evaluated among a total of 83 patients. Humanin and MOTS-c peptide levels were determined in these groups. Nitric oxide and 3-nitrotyrosine were examined to determine nitrosative stress. Superoxide dismutase was included in the study as an antioxidant. Annexin V level was determined by flow cytometry to indicate programmed cell death. Result: Fifty-five thalassemia major patients and the control group (n=28) who were being followed up and treated at the Pediatric Hematology and Oncology Polyclinic of Gaziantep University Faculty of Medicine were included in our study. Gender, age, blood taken from the individuals included in the study: Leukocyte, Neutrophil, Lymphocyte, Thrombocyte, average erythrocyte volume MCV (fl), Hemoglobin (g/dl), Blood Urea Nitrogen (mg/dl), creatinine (mg/dl). , AST (IU/l), ALT (IU/l), LDH (IU/l), ferritin values, Hemoglobin F (HbF, %), Hemoglobin A1 (HbA1, %), Hemoglobin A2 (HbA2, %), results was obtained. In our study, 55% (n=46) of the individuals included in the study were female and 45% (n=37) were male. Compared to the control group, 3-NT levels increased significantly in the thalassemia patient group not receiving chelators. (P<0.05) A significant difference was found when compared to the thalassemia patient group that did not receive chelators. There was no significant difference between the groups in mitochondrial superoxide dismutase (SOD2) results in serum samples (P = 0.4530). When the humanin results in serum samples were compared with the control group, humanin levels decreased significantly in the thalassemia patient groups who received and did not receive chelators (P<0.001). When MOTS-c results in serum samples were compared with the control group, MOTS-c levels were significantly increased in the serum of thalassemia patients who did not receive chelators (P<0.05). When nitric oxide (NO) results were compared with the control group, NO levels were significantly decreased in thalassemia patients not receiving chelators, and a significant difference was detected when compared with the thalassemia patient group not receiving chelators (P<0.01). Compared to the control group, the percentage of apoptosis in the serum of thalassemia patients not receiving chelators increased significantly. A significant difference was detected when compared to the thalassemia patient group not receiving chelators. Conclusion: Annexin V and 3-nitrotyrosine levels were observed to be significantly higher when the patient groups receiving and not receiving thalassemia chelators were compared with the control group. Superoxide dismutase enzyme level was not found to be significantly different between groups. Humanin levels were found to be lower in the patient group not receiving chelators than in those receiving chelators, and MOTS-c was higher in the group not receiving chelators compared to those receiving chelators. Keywords: Thalassemia major, nitrosative stress, apoptosis, ferritin, humanin, MOTS-c, superoxide dismutase, nitric oxide, 3-nitrotyrosine, annexin V