Evaluation of apoptotic proteins (p53 and Bcl-2) expression in trophoblastic tissue of women infected with toxoplasma gondii diagnosed by polymerase chain reactiono

number: 
2344
English
Degree: 
Imprint: 
Medicine
Author: 
Safaa Salman Mezban Al-Thammery
Supervisor: 
Dr. Nidhal Abdul-Muhaimen Mohammed
year: 
2009

Abstract:

A total of fourty women, their age ranged from (20 − 50) years, were enrolled in the current study and were further classified into three categories: • Toxoplasma gondii positive patients: with spontaneous miscarriage
n= 20 women, • Toxoplasma gondii negative patients: with spontaneous miscarriage n= 10 women. • Control group (women with induced abortion for medical causes): n=10 women. From each patient and control, blood as well as trophoplastic tissue were collected. Venous blood was collected from all patients for the detection of specific anti-Toxoplasma gondii IgM in the serum using the ELISA test. polymerase chain reaction technique (PCR) was used for the detection of Toxoplasma gondii DNA in trophoplastic tissue as a diagnostic methods for T.gondii for both positive and negative groups Trophoblastic tissue during curettage was collected and stored as paraffin embedded blocks for both the diagnosis of Toxoplasma gondii B1 gene and for immunohistochemical analysis for the detection of apoptotic proteins (Bcl-2 and p53). The age distribution among positive group patients is between 18 to 41 years old, the results showed that the ages between (27-31) years were the higher group percentage (25.0%). Twelve (40%) samples out of the (30) were negative for anti-Toxoplasma gondii IgM while the rest 18 (60%) were positive for IgM, but when using polymerase chain reaction (PCR) two of the negative samples in ELISA were found to be positive for toxoplasma B1 specific gene. This assures the specificity and sensitivity of polymerase chain reaction as a molecular method for diagnosis of different infectious agents at different body sites. The toxoplasma specific B1 gene was the main target to detect the presence of the parasite in the samples for both toxoplasma gondii positive and negative aborted females by using the polymerase chain reaction (PCR). The highest percentage of Bcl-2 was found in the T.gondii negative group (34.90%±3.44) and control group (38.19%±0.79, while the lowest percentage (10.93%±1.44) was found in the T.gondii positive group. There was no statistical difference (p ≤ 0.363) in the mean percentage of Bcl-2 between T.gondii negative and control group; while there was a high significant differences (p=0.000) in the mean percentage of Bcl-2 in T.gondii positive group compared to cotrols. In addition a high significant differences (p= 0.000) in the mean percentage of Bcl-2 was found between T.gondii positive and negative groups. The highest percentage of expression of p53 protein was found in the T.gondii positive group (31.56%±1.28) and the lowest percentage was in both T.gondii negative and control groups (18.93±1.054 and 11.73±1.810 respectively). There was statistical difference (p ≤ 0.003) in the mean percentage of p53 between T.gondii negative and control groups .In addition there was high significant differences (p=0.000) in the mean percentage of p53 between T.gondii positive group and control groups and between T.gondii positive and negative groups. These high levels of p53 protein found in positive samples for toxoplasma gondii infection might indicated the important role of this protein in cell death and induction of apoptosis that lead to the end of pregnancy with abortion. Low levels of Bcl-2 in aborted women infected with T.gondii might indicated that Bcl-2 have no role in preventing or initiate apoptosis.