Μελέτη μέσω RT-PCR της γονιδιακής έκφρασης της ντεκορίνης σε υποξικοισχαιμικό εγκέφαλο νεογέννητων επιμυών

Abstract

Under hypoxic-ischemic conditions, increased deposition of extracellular matrix proteins occurs in various tissues. The aim of this study is to investigate the effect of hypoxia-ischemia on the decorin gene expression in neonatal rat brain. Seven day old wistar rat pups became ischemic by permanent ligation of the left common carotid. Hypoxia was induced by exposing the pups in a 8% oxygen - 92% nitrogen gas mixture for 60 minutes. At the end of this period and after 2 and 24 hours the animals were decapitated and each cerebral hemisphere was preserved. Total RNA was extracted from cerebral cortex and hippocampus of both cerebral hemispheres. Ten μΐ of total RNA were subjected to reverse transcription using the M-MuLV reverse transcriptase. Five μΐ of the reverse transcription reaction mixture were subjected to competitive PCR amplification under standard contitions, including beta-actin as internal control for gene expression. Decorin mRNA is expressed at comparable levels both in the cerebral cortex and the hippocampus of the neonatal rat brain. Decorin mRNA levels are of limited abudance compared to those of beta-actin. Decorin mRNA is expressed at comparable levels both in the right (hypoxic) and the left (hypoxic - ischemic ) cerebral hemispheres of the neonatal rat brain. Our results suggest that hypoxia - ischemia slightly reduced the levels of decorin mRNA in both cerebral cortex and hippocampus of neonatal rat brain the first 2 hours after , but within 24 hours this effect seemed to be overturned.