| Ischemic stroke is caused by cerebral vascular occlusion or stenosis of cerebral ischemic anoxia,and brain cell death,caused a pathophysiological process of neurologic deficits.Mitochondria is the main place to life activities to provide energy metabolism,is one of the cells in ischemia early accumulated easily organelles,mitochondria in brain ischemia,split the fusion protein of dynamic balance is broken,damaged cause apoptosis and mitochondrial function.Pursuant to the adr is an early use of drugs in clinical treatment of acute cerebral ischemia,has been shown to significantly reduce the cerebral infarction area improve nerve function score,at the same time can reduce the reactive oxygen species(ROS).This experiment was to investigate in accordance with the adr of Drp1 in focal cerebral ischemia in rats and the effect of dynamic change of Mfn2 and brain protection mechanism.Objective:By observing the expression in focal cerebral ischemia rats protein and mRNA of Drp1 and Mfn2.Observe the application of the edaravone in the influence of interference.Discussing the protection of the edaravone in focal cerebral ischemia in rats,and discussing mechanism and the effect of dynamic change of Drp1 and Mfn2,and the detection of edaravone in focal cerebral ischemia.Methods:1.72 male SD rats,according to random number table method is divided into three large groups,there are model group,control group and treatment group.And every large group are depending on the duration of cerebral ischemia of rats were divided into: first day group(n = 8),third day group(n = 8),,seventh day group(n = 8),in total three subgroups.Rats of model group and treatment group line bolt embolism making middle cerebral artery embolism rats model of legal system,the control group rats blood vessels just cut to the neck and neck.Treatment group rats after surgery for 30 minutes to give the adr by intraperitoneal injection of drugs,model group and control group at the same time with the method with the dose of saline solution.Injection in accordance with the adr and physiological saline were injected once a day,medication to take brain tissue.2.Within two hours after the building the MCAO models,adopting Zea Longa method of model group and eda group in cerebral infarction neurological behavioral scores in rats.Scores between 1 to 4 as cerebral infarction model building success,choosing scores in 1to3 rats as experimental object,eliminate neurological deficits overweight or no damage in rats,and to be supplemented,to ensure the number of the rats immutability.3.Observing the pathological changs of rats' brain tissue of each group by hematoxylin-eosin staining technology.4.Detecting the protein expression changes of Drp1 and Mfn2 of ischemic cerebral tissue rats by Western-blot method.5.Detecting the expression level of Drp1 mRNA and Mfn2 mRNA in brain cells of each group by qRT-PCR.Results:1.The results of the neurological defect score is that: the scores of the control group were all 0;the scores of each time point in experimental group respectively were2.80±0.11,2.83±0.12,2.90 ±0.13;the scores of three point in edaravone group were:1.38±0.09,1.40±0.06,1.49±0.08.Compared with the experimental group,the scores in edaravone group were differently decreased(P<0.05).2.HE staining observation experiment of rat brain pathology change: normal control group rats cortex cells form,complete structure,closely packed,have no cerebral cortex damaged nerve cells;Model group rats cortex neurons severely damaged,visible brain cortex,patchy necrosis of neurons shrink,the nucleus pycnosis,hyperchromatic,extend the pathological damage degree aggravating gradually over time;Treatment group at each time point pathology injury model group significantly alleviate nerve cells.3.Western blot method determination of Drp1 in experimental rats with ischemic cerebral cortex cells and Mfn2 protein expression: In the control group the Drp1 protein expression level at each time point respectively is: 1d 0.46±0.03?3d 0.42±0.04?7d 0.41±0.04;In the model group Drp1 protein expression level at each time point respectively is: 1d 0.72±0.06 ? 3d 0.97±0.05?7d 1.57±0.06;In the treatment group the Drp1 protein expression level at each time point respectively is:1d 0.58±0.04,3d 0.73±0.03,7d 0.97±0.05.In the control group the Mfn2 protein expression level at each time point respectively is:1d 0.72±0.02?3d 0.68±0.04?7d 0.69±0.05;In the model group Mfn2 protein expression level at each time point respectively is: 1d 2.18±0.17?3d 1.64±0.14?7d 1.02±0.09;In the treatment group the Mfn2 1 protein expression level at each time point respectively is: 1d 1.64±0.13?3d 0.91±0.09?7d 0.82±0.07.At the same time Drp1 protein in model group is highest,the eda group's expression in lower(p<0.05).At the same conduct,the expression is more and more according to time,and it is reach the peak at the 7d.On the country,the Mfn2 protein expression in the model group is lowest at the same time,he eda group's expression in higher(p<0.05),the sham group's expression is between eda group and model group(p<0.05).At the same conduct,the expression is more and more according to time,and it is reach the low ebb at the 7d(p<0.05).4.qRT-PCR method determination of Drp1 and Mfn2 mRNA in experimental rats with ischemic cerebral cortex cells : in the control group Drp1 mRNA expression level at each time point respectively is:0.72±0.02?0.68±0.04 ? 0.69±0.05;In the model group is: 1.43±0.09 ? 1.59±0.10 ?2.01±0.05;In the treatment group is: 1.23±0.08,1.38±0.09,1.64±0.07.In the control group Mfn2 mRNA expression level at each time point respectively is:1.02±0.04?0.98±0.06?0.96±0.08;In the model group is: 2.06±0.13?1.75±0.07?1.57±0.08;In the treatment group is: 1.76±0.09,1.56±0.10,1.35±0.07.At the same time Drp1 mRNA in model group is highest,the eda group's expression in lower(p<0.05).At the same conduct,the expression is more and more according to time,and it is reach the peak at the 7d.On the country,the Mfn2 mRNA expression in the model group is lowest at the same time,he eda group's expression in higher(p<0.05),the sham group's expression is between eda group and model group(p<0.05).At the same conduct,the expression is more and more according to time,and it is reach the low ebb at the 7d(p<0.05).Conclusion:1.FCI can result in impaired mitochondrial function and destroy the dynamic balance of Drp1 and Mfn2 protein lead to aggravate the brain damage.2.Edaravone of rats with early cerebral infarction have protective effect,the Edaravone mentioned mechanism may be related to mitochondrial damage resistance,to maintain mitochondrial function,less apoptosis,multiply Drp1 expression,reduced Mfn2 expression. |
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