Polymorphism of plasminogen activator inhibitor gene G43A 1 (PAI-1) in patients with acute complications of ulcer disease

Abstract

Objective of the study. To carry out a comparative analysis of the frequency of PAI-1 gene G43A polymorphism in citizens of Chernivtsi region who suffer from various forms of ulcer disease, and to study the possible relationship between different types of genotype and the development of complications.Material and methods. 60 patients with ulcer disease were examined: 42 (70%) men, 18 (30%) women. 37 (61.67%) patients had a duodenal ulcer, the rest ones (38.33%) had a gastric ulcer. Uncomplicated ulcer disease was diagnosed in 12 (20%) patients. 5 (8.33%) patients were found to have an ulcer perforation. In 43 (71.67%) patients there was an ulcer complicated by acute bleeding. In 29 (67.44%) patients the bleeding was stopped. In 14 (32.56%) patients there was a relapse of bleeding.PAI genotyping by G43A mutation was performed by using polymerase chain reaction (PCR). PCR amplification of the corresponding RAI gene fragment was performed by using a specific pair of primers 5’-CCA ACA GAG GAC TCT TGG TC-3’ and 5’- CAC AGA GAG AGT CTG GCC ACG-3’. PCR was performed by using a CFX96 amplifier (Bio-Rad, USA). PCR results were carried out by electrophoresis in 2% agarose gel using a tris-borate buffer. For DNA fragments visualization the gel was stained with ethidium bromide and photographed using ultraviolet light on the GelDoc 2000 installation (BioRad, USA). To determine the length of the obtained fragments, their electrophoretic mobility was compared with the mobility of the DNA marker Gene Ruler DNA Leader Mix (Thermoscientific). The statistical dependence between the values was tested by determining the Fisher criterion, χ2-criterion by Pearson, in particular, the correspondence of the genotype distribution to the Hardy-Weinberg equilibrium.Results. 94.12% patients without acute bleeding and 91.67% patients without acute complications were found to have GG genotype. One female patient (8.33%) without any acute complications was found to have homozygous AA genotype by the mutant allele. No patient with an ulcer perforation was a carrier of allele A. None of the patients without bleeding had a heterozygous GA genotype. GA genotype was found in 25.58% of patients with an acute ulcerative bleeding, which is statistically significantly higher than in patients without any bleeding. Allele A was more common among patients with recurrent bleeding than in ones without it.Conclusions. 1. Among patients with ulcer disease without acute complications of Chernivtsi region 91.67% of whom have homozygous GG genotype by polymorphism G43A РАІ-1 gene, and 8.33% – AA genotype; no heterozygous genotype GA was found in any case.2. All examined patients with a perforated ulcer had the GG genotype.3. Patients with acute ulcerative bleeding, 27.91% of whom are carriers of mutant allele A (among them 25.58% have heterozygous GA genotype, and 2.33% – homozygous AA genotype), which is statistically significantly higher than in patients with ulcer disease without any bleeding and it shows the role of hereditary disorders of the PAI-1 gene in the development of ulcerative hemorrhages. 4. Accounting for G43A polymorphism of PAI-1 gene may become a component of the whole complex for predicting the occurrence of ulcerative bleeding in clinical conditions.