Paraoxonase Gene Polymorphism Analysis in Pediatric Diabetic Patients

Öz

Objective: Identifying gene polymorphisms linked to childhood-onset diabetes mellitus (DM), a multifactorial disease, will illuminate its genetic underpinnings. This study aimed to explore the association between childhood DM and the polymorphisms Paraoxonase1 (PON1) 55 and PON1 192. Method: A total of 129 children who applied to our Child Health and Diseases Endocrinology outpatient clinic participated in this prospective research. The study included 75 children with diabetes, aged 3–15 years (Patient Group-PG), and 54 healthy children (Control Group-CG) as a control. Genotyping of PON1 55 and PON1 192 polymorphisms was performed in study patients using Polymerase Chain Reaction (PCR) and RFLP methods following DNA isolation. Results: Of 54 CG, 21 (38.9%) displayed the homozygous normal (LL) PON1 55 genotypes. The PG contained 25 (33.3%) homozygous normal patients out of 75. In the CG, 25 participants (46.3%) and 39 participants in the PG (52.0%) had the heterozygous (LM) genotype. The homozygous mutant (MM) genotype was observed in 8 (14.8%) of CG and 11 (14.7%) of PG. The CG showed 67 L alleles, representing 62.0% of the total alleles. On the other hand, the number was 89 (59.3%) in the PG. The two groups showed no statistically significant differences in gene polymorphisms, allele frequencies, and M and R allele distribution. Conclusion: No significant relationship was observed between PON1 55 and PON1 192 gene polymorphisms and DM in our study.

Anahtar Kelimeler

• Diabetes mellitus
• DNA Extraction
• Paraoxonase Gene
• Polymerase Chain Reaction
• Polymorphism

Pediatrik Diyabetik Hastalarda Paraoksonaz Gen Polimorfizm Analizi

Öz

Amaç: Çok faktörlü bir hastalık olan çocukluk çağı başlangıçlı diyabet (DM) ile ilişkili gen polimorfizmlerinin belirlenmesi, hastalığın genetik temellerini aydınlatacaktır. Bu çalışma, çocukluk çağı DM ile Paraoksonaz 1 (PON1) 55 ve PON1 192 polimorfizmleri arasındaki ilişkiyi araştırmayı amaçlamaktadır. Yöntemler: Bu prospektif araştırmaya Çocuk Sağlığı ve Hastalıkları Endokrinoloji polikliniğimize başvuran toplam 129 çocuk katılmıştır. Çalışmaya 3-15 yaş arası 75 diyabetli çocuk (Hasta Grubu-HG) ve kontrol grubu olarak 54 sağlıklı çocuk (Kontrol Grubu-KG) dahil edilmiştir. Çalışma hastalarında DNA izolasyonunu takiben Polimeraz Zincir Reaksiyonu (PCR) ve RFLP yöntemleri kullanılarak PON1 55 ve PON1 192 polimorfizmlerinin genotiplendirmesi gerçekleştirildi. Sonuçlar: 54 KG'nin 21'i (%38,9) homozigot normal (LL) PON1 55 genotipini gösterdi. PG, 75 hastanın 25'ini (%33,3) homozigot normal hastadan oluşturdu. KG'de 25 katılımcı (%46,3) ve HG'de 39 katılımcı (%52,0) heterozigot (LM) genotipe sahipti. Homozigot mutant (MM) genotipi, KG'nin 8'inde (%14,8) ve HG'nin 11'inde (%14,7) gözlendi. KG, toplam alellerin %62,0'sini temsil eden 67 L aleli gösterdi. HG'de ise bu sayı 89 (%59,3) idi. İki grup arasında gen polimorfizmleri, alel frekansları ve M ve R alel dağılımı açısından istatistiksel olarak anlamlı bir fark görülmedi. Sonuç: Çalışmamızda PON1 55 ve PON1 192 gen polimorfizmleri ile DM arasında anlamlı bir ilişki gözlenmedi.

Anahtar Kelimeler

• Diabetes mellitus
• DNA Ekstraksiyonu
• Paraoksonaz Geni
• Polimeraz Zincir Reaksiyonu
• Polimorfizm.

Kaynakça

1. 1.Adiga U, Banawalikar N and Menambath DT.Association of paraoxonase 1 activity and insulinresistance models in type 2 diabetes mellitus: Cross-sectional study. J Chin Med Assoc 2022;85:77-80.
2. 2.Besser REJ, Bell KJ, Couper JJ, et al. ISPAD ClinicalPractice Consensus Guidelines 2022: Stages of type1 diabetes in children and adolescents. PediatrDiabetes 2022;23:1175-87.
3. 3.Dada AO, Ikpegbu UA, Okunowo LO, et al. Plasmaparaoxonase-1 activity levels in patients with type 2diabetes mellitus in Lagos State University TeachingHospital, Lagos, Southwest Nigeria: a cross-sectionalstudy. Pan Afr Med J 2023;45:40.
4. 4.El-Lebedy D, Kafoury M, Abd-El Haleem D, et al.Paraoxonase-1 gene Q192R and L55Mpolymorphisms and risk of cardiovascular disease in Egyptian patients with type 2 diabetes mellitus. J Diabetes Metab Disord 2014;13:124.
5. 5. Helaly MAH, Abdel-Khalek EES, Abdel-Hafez HA, et al. Paraoxonase1 55 and 192 gene polymorphisms in an Egyptian population with diabetic complications.Int J Diabetes Dev Ctries 2013;33:207-12.
6. 6.Grzegorzewska AE, Ostromecka K, Adamska P, etal. Paraoxonase 1 gene polymorphisms concerningnon-insulin-dependent diabetes mellitusnephropathy in hemodialysis patients. J DiabetesComplications 2020;34:107687.
7. 7.Sanda GM, Toma L, Barbalata T, et al. Clusterin,paraoxonase 1, and myeloperoxidase alterationsinduce high-density lipoproteins dysfunction andcontribute to peripheral artery disease; aggravationby type 2 diabetes mellitus. Biofactors 2022;48:454-68.
8. 8. Nawaka N, Pansang P, Saniwa A, et al. Paraoxonase1 (PON1) L55M and Q192R polymorphisms are notassociated with chronic kidney disease in Thaiindividuals with type 2 diabetes. Int J Clin Pract2021;75:e1498.

9. 9.Nessler K, Grzybczak R, Nessler M, et al.Associations between myeloperoxidase andparaoxonase-1 and type 2 diabetes in patients withischemic heart disease. BMC Cardiovasc Disord2022;22:521.
10. 10.Libman I, Haynes A, Lyons S, et al. ISPAD ClinicalPractice Consensus Guidelines 2022: Definition,epidemiology, and classification of diabetes inchildren and adolescents. Pediatr Diabetes 2022;23:1160-74.
11. 11.Johns MB, Jr. and Paulus-Thomas JE. Purificationof human genomic DNA from whole blood usingsodium perchlorate in place of phenol. Anal Biochem1989;180:276-8.
12. 12.Humbert R, Adler DA, Disteche CM, et al. Themolecular basis of the human serum paraoxonaseactivity polymorphism. Nat Genet 1993;3:73-6.
13. 13.Adkins S, Gan KN, Mody M, et al. Molecular basisfor the polymorphic forms of human serumparaoxonase/arylesterase: glutamine or arginine atposition 191, for the respective A or B allozymes. AmJ Hum Genet 1993;52:598-608.
14. 14.Akgun O, Duru NS and Elevli M. SerumAntioxidative Enzymes Levels and Oxidative StressProducts in Children and Adolescents with Type IDiabetes Mellitus. J Pediatr Res. 2018;5:128-34.
15. 15.Abudayyak M, Boran T, Tukel R, et al. The Role ofPON1 Variants in Disease Susceptibility in a TurkishPopulation. Glob Med Genet. 2020;7:41-6.
16. 16.Zhou M, Liu X-H, Liu Q-Q, et al. Lactonase activityand status of paraoxonase 1 and oxidative stress inneonates of women with gestational diabetesmellitus. Pediatr Res. 2021;89:1192-9.
17. 17.Yari A, Karam ZM, Meybodi SM, et al. CDKN2B‐AS(rs2891168), SOD2 (rs4880), and PON1 (rs662)polymorphisms and susceptibility to coronaryartery disease and type 2 diabetes mellitus inIranian patients: A case‐control study. Health SciRep. 2023;6:e1717.