Lack of Association between Angiotensin Converting Enzyme I/D Polymorphism and Unexplained Recurrent Miscarriage in Saudi Arabia Translated title: NEPOSTOJANJE POVEZANOSTI IZMEĐU I/D POLIMORFIZMA ANGIOTENZIN-KONVERTUJUĆEG ENZIMA I NEOBJAŠNJENIH VIŠESTRUKIH SPONTANIH POBAĆAJA U SAUDIJSKOJ ARABIJI

Summary

Background

An insertion/deletion (I/D) polymorphism in the angiotensin converting enzyme (ACE) gene has been associated with recurrent miscarriage (RM) in several populations. We initiated this study to determine the association, if any, between the I/D polymorphism of ACE gene and RM in Saudi females.

Method

This study was conducted on 61 Saudi females suffering from RM (mean age: 34.1±6.2 years; range 15–45) attending clinics at King Khalid University Hospital, and 59 age matched females who had at least 2 children, as controls. Blood samples were drawn in EDTA tubes by venipuncture. DNA was extracted using the Puregene DNA purification kits. Insertion/Deletion (I/D) polymorphism of ACE gene was investigated by amplifying the genomic DNA by PCR using gene-specific primers. A single 190 bp or 490 bp band was obtained in the homozygous cases for the D allele or I allele, respectively, while the presence of both 190 and 490 bp bands indicated heterozygosity (ID).

Statistical analysis

Deviation from Hardy-Weinberg equilibrium was determined ( http://ihg.gsf.de/cgi-bin/hw/hwa1.pl). A standard chi-square (χ ²) test was used for comparing the genotype and allele frequencies in the two groups and Students’t’ test and χ ² test were employed to compare values between the two groups. P<0.05 was considered statistically significant.

Results

The frequencies of DD, ID, and II genotypes were 56.7%, 29.5% and 4.9%, respectively, in females with RM and 54.2%, 42.3% and 3.3% respectively in the control group, but the difference was not statistically significant.

Conclusion

In some populations, meta-analyses showed an association between I/D polymorphism and RM risk, and the D allele was implicated as an increased risk factor for RM. However, this association was not apparent in the Saudi females.

Kratak sadržaj

Uvod

Insercioni/delecijski polimorfizam (I/D) u genu za AKE (angiotenzin-konvertujući enzim) doveden je u vezu sa višestrukim spontanim pobačajima (VSP) u nekoliko populacija. Ovu studiju smo sproveli kako bismo utvrdili da li postoji ili ne postoji povezanost između I/D polimorfizma gena za AKE i VSP kod žena u Saudijskoj Arabiji.

Metod

Studija je obuhvatila 65 žena saudijske nacionalnosti sa VSP (prosek godina: 34,1±6,2 godina; raspon 15–45) koje su se lečile na klinikama Univerzitetske bolnice kralj Halid i 65 žena iste starosne dobi koje su imale najmanje dvoje dece, kao kontrolnu grupu. Uzorci krvi sakupljani su u EDTA epruvete venepunkcijom. DNK je ekstrahovana pomoću Puregene DNA purification kitova. Insercioni/delecijski (I/D) polimorfizam gena za AKE je ispitivan putem amplifikacije genomske DNK pomoću PCR uz korišćenje prajmera specifičnih za gene. U slučaju homozigota za D alel ili I alel dobijana je po jedna traka duga 190 bp ili 490 bp, dok je prisustvo obe trake od 190 i 490 bp značilo heterozigotnost (ID).

Statistička analiza

Procenjeno je odstupanje od Hardi-Vajnbergove ravnoteže. Za poređenje učestalosti genotipova i alela u dve grupe korišćen je standardni hi-kvadratni test (c2), dok su za poređenje vrednosti između dve grupe primenjeni studentov T test i c2 test. P < 0,05 smatrano je statistički značajnim.

Rezultati

Učestalosti DD, ID i II genotipova bile su 56,7%, 29,5% i 4,9% kod žena sa VSP, odnosno 54,2%, 42,3% i 3,3% u kontrolnoj grupi, ali razlika nije bila statistički značajna.

Zaključak

U nekim populacijama metaanalize su pokazale povezanost između I/D polimorfizma i rizika za VSP, ali D alel je bio umešan kao faktor povećanog rizika za VSP. Međutim, ova povezanost nije bila ispoljena kod saudijskih žena.