Abstract | Mutacije u genu za metilentetrahidrofolat-reduktazu (MTHFR), C677T i A1298C, uzrokuju smanjenu funkcionalnost i aktivnost MTHFR-a, što zbog nakupljanja metaboličkog nusprodukta homocisteina u stanicama uzrokuje stanje oksidativnog stresa i pokreće niz patoloških procesa koji dovode do hiperkoagulabilnog stanja. Dosadašnja istraživanja mutacija u genu za MTHFR i homocisteina, u ulozi rizičnog čimbenika venskih tromboembolijskih bolesti, kao što je plućna embolija (PE), ishodila su neusklađene rezultate zbog interakcije velikog broja obuhvaćenih činitelja. Cilj ovog istraživanja bio je ispitati povezanost dviju mutacija u genu za MTHFR s rizikom od razvoja PE-a na uzorku hrvatske populacije. U studiju istraživanja parova uključeno je 110 pacijenata s dijagnozom PE-a te 101 dobrovoljni davatelj krvi koji čine kontrolnu skupinu. Izolacija genomske DNA iz uzorka pune krvi učinjena je pomoću komercijalnog kompleta reagencija QIAamp DNA Blood mini QIAcube kit na uređaju QIAcube (Qiagen, Njemačka). Za genotipizaciju mutacija MTHFR-a, C677T i A1298C, metodom lančane reakcije polimerazom u stvarnom vremenu korišten je uređaj ABI Prism 7500 Real-time PCR system (Applied Biosystems, SAD). Dobiveni rezultati analizirani su χ2-testom i testom razlika proporcija uz 95 % intervale pouzdanosti (CI) i razinu statističke značajnosti od 0,05 (p-vrijednost). Usporedbom zastupljenosti pojedinih genotipova MTHFR-a za C677T nije dobivena statistički značajna razlika između skupine pacijenata s PE-om i kontrolne skupine (χ2= 0,17; p=0,921), kao ni za mutaciju A1298C (χ2= 0,19; p=0,909). Također, ni učestalosti mutiranog alela nisu se statistički značajno razlikovale kod bolesnika i kontrolnih ispitanika. Usporedbom kombinacija genotipova C677T i A1298C ispitanika između skupina, najveće razlike dobivene su za kombinacije CC, AA (3,6 %) te CC, CC (1,8 %). Međutim, test razlike proporcija potvrdio je da navedene razlike nisu statistički značajne (p= 0,514; p=0,687). Rezultati ove studije pokazali su da na ispitanom uzorku hrvatske populacije, mutacije u genu za MTHFR: C677T i A1298C, ne predstavljaju rizični čimbenik za razvoj plućne embolije. |
Abstract (english) | MTHFR gene mutations, C677T and A1298C, reduce enzyme activity and decrease its function which leads to the accumulation of potentialy toxic metabolite homocysteine. The consequence is increased oxidative stress and other pathological processes that provide the state of hipercoagulability. Recent studies on mutations in MTHFR gene and homocysteine in a role of risk factor for venous thromboembolic disorders, including pulmonary embolism, have reported inconsistent results because of interaction of numerous factors. The aim of this study was to examine relationship between two mutations in the MTHFR gene and the risk of pulmonary embolism. This case-control study has included 110 patients with pulmonary embolism and 101 healthy controls (blood donors). Genome DNA from whole blood samples was isolated via commercial kit, QIAamp DNA Blood mini QIAcube kit on QIAcube device (Qiagen, Germany). The genotyping of MTHFR mutations C677T and A1298C was performed using real time polymerase chain reaction on the analyzer ABI Prism 7500 Real-time PCR system (Applied Biosystems, USA). Data was tested using statistical methods, χ2-test and comparison of proportions, with confidence intervals of 95 % and the value of statistical significance of 0,05 (p-value). There was no statistically significant difference in presence of the MTHFR C677T genotypes among patients with PE and controls (χ2= 0,17; p=0,921). Same results have been observed for the A1298C mutation (χ2= 0,19; p=0,909). The data regarding the frequency of mutated allele in PE group and control group also showed no singificant association with PE. When the combinations of MTHFR C677T and A1298C genotypes were compared, greatest differences between groups have been reported for CC, AA (3,6 %) and CC, CC (1,8 %) combinations. Even though, there was no statistically significant difference between frequencies of the two groups (p= 0,514; p=0,687). The results of this study demonstrated that MTHFR mutations: C677T and A1298C, does not represent a risk factor for the development of pulmonary embolism. |