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Abstract
Pre-eclampsia affects 2–7% of all pregnancies with varying severity and is a leading cause of maternal and fetal mortality and morbidity. Observations have led to speculations regarding an immunological basis or component in pre-eclampsia because previous exposure to foreign or paternal antigens appears to reduce the risk of pre-eclampsia.
HLA-G is a specialized antigen expressed only by trophoblast cells and specifically binds to inhibitory receptors expressed by decidual leukocytes to abrogate activating signals received by these cells.
The aim of this study was to assess the expression of HLA-G mRNA in the placental tissue of preeclamptic versus normal pregnant women.
And to analyze the correlation between the level of HLA-G expression and the different parameters defining pregnancy outcome.
This study comprised two groups of primigravid females at labour: Group 1 (Preeclamptic Group) consisting of 20 preeclamptic pregnant females at the age of 20- 40 years and Group 2 (Control Group)consisting of 20 normal healthy pregnant females who matched the patient group for age. All subjects were not suffering from concurrent medical illness as diabetes or autoimmune diseases or receiving any medications. Data concerning gestational age, rout of delivery, placental weight, fetal weight, fetal sex, and apgar score were recorded for each subject of this study.
Placental tissue samples were obtained from each subject of both groups and were used for isolation of total mRNA using a spin total RNA isolation system of Promega corporation,Madison,USA. mRNA was subjected to reverse transcription followed by polymerase chain reaction
(RT-PCR) using primers specific for HLA-G gene and primers specific for β-actin. The primers were designed using the computer program: Accelrys Discovery Studio Gene 1.5 (Accelrys DS Gene 1.5)and co-amplification of HLA-G gene with β-actin as an internal standard was performed using QIAGEN OneStep RT-PCR Kit( reverse transcription for 30 min at 50οC followed by initial PCR activation step for 15 min at 95 οC,then 35 PCR cycles: denaturation for 1 minute at 95 οC, annealing for 1 minute at 50 οC, and extention at 72°C for 1 minute, followed by final extension for10minutes at 72°C.