Placental Leucine Aminopeptidase/Oxytocinase Expression in Miscarriage
Taghreed H. AL Sadoon *
Department of Biochemistry, College of Medicine, University of Tikrit, Iraq.
Ferah Gh. Al-Salihi
Department of Chemistry, College of Sciences, University of Tikrit, Iraq.
Adnan I. Al-Badran
Department of Biology, Cell and Biotechnology Research Unit, College of Sciences, University of Basrah, Iraq.
*Author to whom correspondence should be addressed.
Abstract
Objectives: To evaluate the role of placental leucine aminopeptidase (P-LAP) in miscarriage and searching to select the gene of this enzyme in miscarriage to exclude direct or indirect effects of abortion.
Methods: Total RNA is purified from the fresh placental tissue sample according to the protocol of the QIAamp®RNA Tissues Mini Kit (Qiagen). The sample is visualized by 0.7% agrose gel containing. Total RNA is reverse-transcribed RT-PCR carried out on the previously extracted RNA samples using (QIAGEN One Step RT –PCR Kit). Aliquots (1ml) of RT reaction samples are amplified by PCR, the PCR products (10µl) per lane are resolved by using electrophoresis on 2.0% agrose gel .Later the product is detected and examined under UV transiluminator.
Results: The obtained results indicate that there is an expression of P-LAP mRNAs in placentas of normal pregnancy women (28±SE0.45) and there is a decrease in miscarriage. Furthermore, there is a difference between single miscarriage and those with recurrent miscarriage, the mean values in single miscarriage (1st and 2nd) are (16.1%±SE0.84) and (13%±SE0.24) respectively while the mean values in recurrent miscarriage (1st and 2nd) are (12%±SE0.19) and (9%±SE0.52) respectively. The expression of P-LAP is significantly lower (p=0.05) in miscarriage and highly significant decrease in recurrent than in single miscarriage women.
Conclusion: The decrease of P-LAP mRNA levels in miscarriage placentas could be a sign that p-LAP function is an important step in pregnancy regulation.
Keywords: Placental leucine aminopeptidase (P-LAP), complementary deoxyribonucleic acid (cDNA), β-actin as the internal control, multiplex-single step-(M-SS)-RT-PCR assay, electrophoresis.