| Objective:The dynamic expression of ALDH2 mRNA and 4-HNE and the effect of Edaravone on the expression of ALDH2 mRNA in cerebral infarction tissue(rats)were observed.To study the possible mechanism of self-protection of brain tissue after infarction and the possible protective mechanism of Edaravone on ischemic brain tissue.Methods:1 72 healthy male Sprague-Dawley rats weighing 250-330 g were randomly divided into sham operation control group(n=24),embolic experimental group(n=24)and Edaravone group(n=24).Rats in each group were divided into 3 subgroups according to the time of death:1d(n=8),3d(n=8)and 7d(n=8).The rats in the sham operation control group were treated with incision and dissection,that is,the artery was separated but the carotid artery was not ligated,only the surgical trauma was given;the experimental group and Edaravone group made permanent MCAO model,and Edaravone group received intraperitoneal injection of Edaravone half an hour after operation.The sham operation control group and the embolization group were given the same amount of sodium chloride solution half an hour after operation.2 After 2 hours of fish line embolization,neurobehavioral scores of cerebral infarction rats in experimental group and Edaravone group were evaluated by Zea Longa method.The successful rats with scores of 1-3 were selected as the experimental objects in this experiment,and the number of the experimental objects in each group was the same at the same time.3 The pathological changes of brain tissue in each group were observed by HE staining.4 The relative expression of ALDH2 mRNA in brain tissue of rats in each group was detected by RT-PCR method5 The relative expression of ALDH2 protein in brain tissue of rats with cerebral infarction was calculated by Western blot method.6 Determination of 4-HNE content in brain tissue of rats in each group by ELISE methodResults:1 The pathological changes of brain tissue in different groups of rats were observed by HE staining: the structure and morphology of brain cells in the control group were normal,the brain cells were arranged closely with the peripheral cells,and the nerve cells in the experimental group were obviously necrosed and disappeared.Irregular arrangement,cytoplasm concentrated red stain,deep staining of nucleus,pyknosis,partial dissolving disappeared,this change was the most serious in the group of one day and seven days,and the degree of cell damage in the treatment group at each time point was lighter than that in the experimental group.2 The dynamic changes of 4-HNE in brain tissue of rats in each group were detected by ELISE method.The 4-HNE content in the control group at each time point of 7d,3d and 1d were:110.50±1.67,110.16±5.37,112.88 ± 6.14;In the experimental group,the 4-HNE content at each time point of 7d,3d and 1d were respectively:350.24±5.50,379.12±10.62,441.70±17.58;The contents of 4-HNE in Edaravone group at each time point of 7d,3d and 1d were respe ctively:205.16±5.19,259.43±9.71,326.20±10.66.The content of 4-HNE in bra in tissue of experimental group was significantly higher than that of Edara vone group andcontrol group at the same time point(P<0.01),the content of 4-HNE inbrain tissue of Edaravone group was significantly higher than that of control group at the same time point(P<0.01),4-HNE in experimental group and drug group was significantly higher than that in control group(P<0.01).The content of 4-HNE decreased with time(P<0.01).There was no significant change of 4-HNE content in brain tissue of the controlgroup(P>0.05).3 The relative expression of ALDH2 in brain cells of rats in each group was detected by Western blot method.The expression level of ALDH2 protein in the control group was determined at each time point of 7d,3d and 1d were:0.481±0.010,0.481±0.015,0.484±0.008;In the experimental group,the expression levels of ALDH2 protein in 7d,3d and 1d were:0.710±0.001,0.631 ±0.003,0.541±0.002;in the Edaravone group,the expression level of ALDH2 protein at each point of 7d,3d,1d were 0.842± 0.005,0.791±0.002,0.739± 0.0 01.There was no significant change in the content of ALDH2 protein in each subgroup of the control group(P>0.05),but the content of ALDH2 protein in the experimental group and Edaravone group increased gradually with the prolongation of the model making time(P<0.01).Compared with the control group,the expression of ALDH2 protein in the control group and Edaravone group was significantly increased at each time point(P<0.01),and the expression of ALDH2 protein in the Edaravone group was significantly higher than that in the experimental group at each time point(P< 0.01).4 The dynamic expression of ALDH2 mRNA was as follows:in the control group,the relative expression of ALDH2 mRNA in brain cells of rats with cerebral infarction at different time points of 7d,3d and 1d were:1.02 ± 0.03,1.02±0.06,1.04±0.07;in the experimental group,the expression level of ALDH2 mRNA at different time points of 7d,3d and 1d were:2.14±0.03,1.67 ±0.06,1.29±0.05;in the Edaravone group,the expression level of ALDH2 mRNA at different time points of 7d,3d and 1d were:2.79±0.07,2.37±0.15,1.82±0.08.There was no significant difference in the dynamic expression of ALDH2 mRNA in the control group(P>0.05).The expression of ALDH2 mRNA in the experimental group and Edaravone group increased gradually with the prolongation of the model making time(P<0.01),and compared with the control group,the expression of ALDH2 mRNA in the experimental group and Edaravone group increased gradually(P<0.01).The relative expression of ALDH2 mRNA in the experimental group and Edaravone group was significantly increased at each time point(P<0.01).At the same time,the expression of ALDH2 mRNA in Edaravone group was significantly higher than that in the experimental group(P<0.01).Conclusion:1 Oxidative stress was induced after cerebral infarction in rats,and the endogenous protective mechanism of ALDH2 may play a role in this mechanism.2 Edaravone can scavenge oxygen free radicals and protect neurons against oxidative stress,and its mechanism may be related to increasing the expression of ALDH2 protein and enhancing the antioxidant stress of the body. |
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