Effects Of Phosphocreatine On C-fos Gene Expression In The Hippocampus Of Rats With Cerebral Ischemia Reperfusion Injury

Objective:Established the model of cerebral ischemia-reperfusion injury, to observe theprotective effect of Phosphocreatine on brain cell damage in rats, through detected theexpression of immediate gene c-fosmRNA gene in hippocampus of rats, to investigatethe damage of phosphocreatine on cerebral protective effect and mechanism.Methods: This experiment selected weight between300-350g in27healthy adultmale Sprague-Dawley rats (SD) as the research subjects, were randomly grouping:1.、Sham group (sham group, n=9): After anesthesia(10%chloral hydrate,350mg kg-1injected intraperitoneally）and neck incision,isolated bilateral common carotid arterywithout occlusion, exposure to30min；2、Ischemic reperfusion group (I/R group,n=9),after anesthesia, separation and occlusion of bilateral common carotid artery30min,loosen the artery clamp then reperfusion24h.3、Drug treatment group,(Durg group,n=9),Separated and occlusion of bilateral common carotid artery30min，slowly from thetail vein injection of phosphocreatin（ePCr，80mg.100g-1)，and then reperfusion24h. Theabove three groups of rats were decapitated directly after put cages24h, rapidcraniotomy on the ice to take hippocampus, after the left hippocampus were frozen inliquid nitrogen into the-80℃refrigeratorfor reverse transcription polymerase chainreaction (RT-PCR) detection of c-fos mRNA,the right hippocampus placed in4%formaldehyde saved for immunohistochemical detection.Results: The two rats died of respiratory failure, the rest were through the observationperiod. Durg group of rats mental state better than rats Sham group and strongvitality.1、HE staining results compare：Sham group nerve cells were arranged closely、rules, form and structure of normal；Compared with the Sham group，I/R group ofneurons arranged in scattered， cells structural were damaged, nuclear fragmentation、nuclear condensation; Compared with the I/R group, a large number of nerve cells inDrug group rats has completly structure, arranged in order.2、 Immunohistochemicalstaining for c-fos gene expression results compare: C-fos mRNA showed Brown immunoreactive positive cells in hippocampus of Sham group only a smallamount of expression, pale staining; I/R group hippocampal tissue stained darker andhigh expression levels; Hippocampus c-fos mRNA in Drug group rats has beenexpressed in the nucleus, compared with the I/R group, to reduce staining.3、 RT-PCRto detect the result compare：electrophoresis was no significant difference in threegroups of β-actin stripe width and brightness.I/R group of c-fos electrophoresis stripwidth and thickness, bright；Sham group of c-fos mRNA bands vague, dim；Drug groupof bands in between brightness. The results of the gel image analysis system: I/R groupc-fos/β-action ratio is greater than the Durg group (p <0.05); Durg Group c-fos/β-actionratio is greater than the Sham group (p <0.05).Conclusion:1、Brain damage of cerebral ischemia reperfusion can increase theexpression of c-fosmRNA in nerve cells;2、Phosphocreatine can protect brain cells,decreasing the expression of c-fos mRN；3、C-fosmRNA can be used as an indicator forearly diagnosis of brain damage.