| Objective:To research the possible mechanisms of self-protection against focal cerebral ischemia(FCI)and the possible mechanisms of edaravone against FCI by observing the expression of PRX2 mRNA,PRX6 mRNA and PRX6 protein in cerebral cortex of rats with FCI and edaravone conditioning.Methods:1 72 male Sprague Dawley rats were randomly divided into three groups: sham,model groups and edaravone-treated group(eda group),and according to the different time points each group was divided into three subgroups: 1st day subgroup,3rd day subgroup and 7th day subgroup(n=8 each).The pMCAO models were established in model and eda group rats by intraluminal suture technique,and the sham group rats were just given the separation of the carotid artery.In eda group,edaravone was injected intraperitoneally after 30 mins from pMCAO.The equal volume saline was given to rats in sham and model groups after 30 mins from pMCAO.Giving the daily medicine to rats in each group by injecting intraperitoneally until to death.2 The neurological deficit of FCI rats in model group and eda group were scored by Zea Longa at 2h after pMCAO.1-4 grade for successful model.Selecting the rats scored in 1-3 grade as the experimental animal model,excluding the rats with heavy nerve injury or without injury,and being added.3 Observing the pathological changs of rats' brain tissue of each group by HE.4 Detecting the expression level of PRX6 protein in brain tissue of each group by Western method.5 The expression level of PRX2 m RNA and PRX6 mRNA in brain cells of each group were evalueted by qRT-PCR.6 Detecting the MDA content in brain tissue of each group by the method of TBA.Results:1 The pathological morphological changes of brain tissue in each group rats were observed by HE staining: the cerebral cortex cells in sham group were normal,having tight,clear structure,without damage of cerebral cortex cells;but cerebral cortex cells of model group rats were seriously damaged,with cell shrinkage or nuclear condensation.The pathological damage gradually increased with prolongation of molding time;Compared with the model group rats,the pathological damage of eda group rats were obviously improved.2 The MDA content were detected by the method of TBA: The sham group results in first day,third day and seventh day respectively were 3.938±0.164?3.973±0.211?3.987±0.247;The model group results in first day,third day and seventh day respectively were 12.637±0.595?8.985±0.362?6.762±0.243;The eda group results in first day,third day and seventh day respectively were 9.432±0.299?6.700±0.182?5.191±0.259.The contents of MDA in sham group had no significant difference at different time point(P>0.05).Compared with the sham group,the content of MDA in model and eda group increased significantly(P<0.05).The MDA content in eda group were lower than that in model group,but higher than that in sham group at different time point(P<0.05).With prolongation of molding time,the content of MDA in model and eda groups decreased gradually(P<0.05).3 The PRX2 mRNA and PRX6 m RNA expression levels were observed by qRT-PCR in brain cells: The results of expression level of PRX2 mRNA : The sham group results of PRX2 mRNA in first day,third day and seventh day respectively were 0.990±0.018?0.987±0.026?1.018±0.028.The model group results in first day,third day and seventh day respectively were 1.190±0.018?1.510±0.023?1.820±0.021.The eda group results in first day,third day and seventh day respectively were 1.765±0.016?2.093±0.026?2.438±0.030.The resluts of expression level of PRX6 mRNA: The sham group results of PRX6 mRNA in first day,third day and seventh day respectively were 0.998 ±0.017?1.010±0.020?0.988±0.019.The model group results in first day,third day and seventh day respectively were 1.091±0.015?1.268±0.032?1.397±0.016.The eda group results in first day,third day and seventh day respectively were 1.301±0.038 ? 1.404±0.024 ? 1.509±0.031.With prolongation of molding time,the expression levels of PRX2 mRNA and PRX6 mRNA had no significant difference(P>0.05).The trend of PRX2 mRNA and PRX6 mRNA expression levels were similar.With prolongation of molding time,The expression levels of both in model and eda group were significantly enhanced(P<0.05).Compared with sham group,the expression levels of PRX2 and PRX6 mRNA in model and eda groups were enhanced obviously at different time point(P<0.05).Compared with model group,the expression levels of PRX2 mRNA and PRX6 mRNA in eda group were enhanced obviously at different time point(P<0.05).4 The PRX6 protein expression were detected by Western method in each group: The sham group results of PRX6 protein in first day,third day and seventh day respectively were 0.575±0.011?0.571±0.012?0.583±0.009.The model group results in first day,third day and seventh day respectively were 0.644±0.009?0.858±0.029?1.085±0.034.The eda results in first day,third day and seventh day respectively were 0.848±0.032?1.189±0.047?1.386±0.035.The expression levels of PRX6 protein in sham group had no significant difference at different time piont(P>0.05).With prolongation of molding time,the expression levels of PRX6 protein in model and eda groups were increased gradually(P<0.05).Compared with the sham group,the expression levels of the PRX6 protein in model and eda groups were significantly increased at different time point(P<0.05).Compared with the model group,the expression levels of the PRX6 protein in eda group were significantly increased at different time point(P<0.05).Conclusion:1 FCI can start oxidative-stress response,and PRX2,PRX6 may play an important role in inhibiting the oxidative-stress injury in the process of focal cerebral ischemia.2 Edaravone may play a neuroprotective effect by ameliorating oxidative stress injury and enhancing PRX2 and PRX6 expression on focal cerebral ischemic rats. |
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