The content of ATP Synthase in Cerebral Ischemia of Varying Severity Comparative Analysis

Bon Elizaveta I., Maksimovich Natalia E., Karnyushko Olga A., Zimatkin Sergey M, Lychkovskaya Maria A
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Abstract

Objective. Evaluation of changes in the content of ATP synthase in the parietal cortex and hippocampus of the brain of rats with ischemia of varying severity in a comparative aspect. Methods. The experiments were performed on 88 male outbred white rats weighing 260 ± 20 g. Brain ischemia was modeled under conditions of intravenous thiopental anesthesia (40-50 mg / kg). Total cerebral ischemia was modeled by decapitation of animals. The brain sampling was carried out 1 hour and 24 hours after decapitation - to study tissue respiration of mitochondria, as well as 1 hour later to determine the content of ATP synthase. Subtotal cerebral ischemia was modeled by simultaneous ligation of both common carotid arteries. The material was taken after 1 hour to determine the content of ATP synthase. Stepwise subtotal cerebral ischemia was performed by sequential ligation of both common carotid arteries with an interval of 7 days. The sampling was carried out 1 hour after ligation of the second common carotid artery in each of the subgroups. Partial cerebral ischemia was modeled by ligation of one common carotid artery on the right. The sampling was carried out 1 hour after the operation. Determination of the content of ATP synthase was carried out by immunohistochemical method using monoclonal antibodies. For this purpose, after decapitation, the brain was quickly removed from the rats, pieces of the cerebral cortex were fixed in zinc-ethanol-formaldehyde at + 4 ° C (overnight), then embeddedвinвparaffin. Results. In the group of stepwise subtotal cerebral ischemia, the smallest decrease in the content of ATP synthase was observed in the 1st subgroup with an interval between dressings of 7 days, while the greatest decrease in the content of the enzyme was noted in the 3rd subgroup with the minimum interval between the dressings of the common carotid artery (1 day). Modeling of more severe types of ischemic damage led to pronounced morphological changes in neurons in the parietal cortex and hippocampus of the rat brain - a decrease in their size, deformation of the perikarya, an increase in the degree of neuronal chromatophilia with their simultaneous wrinkling and subsequent death. These disorders were most pronounced in the 3rd subgroup of stepwise subtotal cerebral ischemia with the shortest interval between dressings, which was 1 day, and in the group of total cerebral ischemia. Conclusion. Thus, the most pronounced decrease in the content of ATP synthase was observed in the groups of total cerebral ischemia, subtotal cerebral ischemia and in the 3rd subgroup of stepwise subtotal cerebral ischemia, with a minimal time interval between the ligation of the common carotid artery. In stepwise subtotal cerebral ischemia with an interval between ligation of the common carotid artery of 7 days, the suppression of the ATP synthase content was not so significant.
不同严重程度脑缺血ATP合酶含量的比较分析
目标。比较评价不同程度缺血大鼠脑顶叶皮层和海马ATP合酶含量的变化。方法。实验以体重260±20 g的雄性远交系大鼠88只为实验对象。在静脉注射硫喷妥钠麻醉(40 ~ 50 mg / kg)条件下建立脑缺血模型。采用断头法模拟全脑缺血模型。分别在断头后1小时和24小时采集脑组织样本,研究线粒体的组织呼吸作用,1小时后采集脑组织样本测定ATP合酶的含量。同时结扎双颈总动脉,模拟脑次全缺血。1小时后取液,测定ATP合酶的含量。通过连续结扎双颈总动脉进行逐步脑次全缺血,间隔7天。在每个亚组的第二总颈动脉结扎1小时后进行采样。结扎右侧颈总动脉,建立脑局部缺血模型。术后1小时取样。单克隆抗体免疫组化法测定ATP合酶含量。为此,在斩首后,迅速从大鼠身上取出大脑,将大脑皮层碎片在+ 4°C(过夜)的锌-乙醇-甲醛中固定,然后embeddedвinвparaffin。结果。在逐步次全脑缺血组中,ATP合酶含量下降幅度最小的是第1亚组,间隔7天;而ATP合酶含量下降幅度最大的是第3亚组,间隔时间最短,间隔1天。对更严重类型的缺血性损伤的建模导致大鼠脑顶叶皮层和海马神经元的明显形态学变化——它们的大小减小,核周变形,神经元嗜色性程度增加,同时起皱并随后死亡。这些障碍在逐步次全脑缺血的第3亚组和全脑缺血组中最为明显,间隔时间最短,为1天。结论。因此,在脑全缺血、脑次全缺血组和逐步脑次全缺血的第三亚组中,ATP合酶含量的下降最为明显,结扎颈总动脉之间的时间间隔最短。在颈总动脉结扎间隔7天的逐步次全脑缺血中,ATP合酶含量的抑制不明显。
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