Study of apoptosis and related proteins, CRH and hpGH in placentas of newborns small for gestational age (SGA).

UNLABELLED: The aim of this study is to quantify the expression of apoptotic genes and genes related to the development and growth in placentas of pregnancies with IUGR (intrauterine growth restriction) and normal placentas. We studied the expression of Bcl-2, Caspase3, hpGH (human placental growth hormone) and CRH (corticotropin releasing hormone) genes in normal and IUGR placentas. In addition we have demonstrated this expression by immunohistochemical techniques.

MATERIAL AND

METHODS

Placentas of newborns with intrauterine infections, complicated pregnancies, congenital malformations and birth asphyxia were excluded. RNA extraction and purification. Total RNA was extracted and cleansed from the lysate using RNA wiz (Ambion) and Qiagen Rneasy Mini (Qiagen). cDNA synthesis. This assay was performed using the Retroscript Kit, Ambion. RT-PCR was performed with the LightCycler System 2.0 (Roche Diagnostics) and the LightCycler FastStart DNA Master Plus SYBR Green I. An immunochemical study was performed using the Envision Plus Dako protocol.

RESULTS

Bcl-2, hpGH and CRH are down regulated in the SGA group in comparison to the control group. Caspase3 is up regulated in the SGA group in comparison to the control group. We demonstrated that in placentas from pregnancies with IUGR, expression of Bcl-2is diminished and expression of caspase 3 is augmented compared to normal placentas.