亚ERK5在大鼠脑出血模型中神经细胞凋亡的作用

朱小琴; 赵丹; 杜鹃; 胡东懿

亚ERK5在大鼠脑出血模型中神经细胞凋亡的作用

The role of subERK5 in the neuronal apoptosis in intracerebral hemorrhage model rats

摘要

摘要:
目的： 探究细胞外信号调节激酶5（extracellular signal regulated kinase，ERK5）信号通路在脑出血大鼠模型神经细胞凋亡的作用机制。
方法： 利用自体血注射脑尾状核的方法构建脑出血模型，根据干预手段将大鼠分成模型组、空白对照组、假手术组和实验组，模型组：自体血注射脑尾状核的方法构建脑出血模型。空白对照组：SD大鼠未做任何处理。假手术组：利用上面的方法进行常规处理，但未在尾状核注射自体血，其余同脑出血模型组。实验组：在尾状核注射大鼠自体血后注入BIX02188试剂，其余同模型组大鼠处理。构建模型后6h进行神经功能评分确认。利用RT−PCR和Western blot检测ERK5、Caspase−3和Caspase−9 mRNA和蛋白相对表达水平，利用TUNEL染色检测脑出血模型的神经细胞凋亡。
结果： 实验组神经功能评分高于模型组（P ＜ 0.01），低于空白对照组和假手术组（P ＜ 0.01）。空白对照组、模型组、假手术组和实验组的ERK5、Caspase−3和Caspase−9 mRNA的相对表达量的比较差异具有统计学意义（P ＜ 0.05）。实验组ERK5、Caspase−3和Caspase−9 mRNA的相对表达量高于空白对照组和假手术组低于模型组（P ＜ 0.05）。实验组ERK5、Caspase−3和Caspase−9的蛋白相对表达量低于模型组（P ＜ 0.01），高于空白对照组和假手术组（P ＜ 0.05）。TUNEL染色检测，实验组凋亡指数低于模型组（P ＜ 0.05），高于假手术组和空白对照组（P ＜ 0.01）。
结论： ERK5信号通路可能通过调节Caspase−3和Caspase−9的表达水平介导细胞凋亡，下调ERK5蛋白表达水平可能对神经细胞有保护作用。

Abstract:
Objective To explore the action mechanism of extracellular signal regulated kinase 5(ERK5) signaling pathway in neuronal apoptosis in rat models of intracerebral hemorrhage.
Methods The rat model of cerebral hemorrhage was established by injecting autologous blood into caudate nucleus of brain. According to the intervention methods, the rats were divided into the blank control group, sham operation group, model group and experimental group. The blank control group did not do any treatment. The sham operation group were treated with routine method, but autologous blood was not injected into caudate nucleus. The experimental group were injected with autologous blood into caudate nucleus. The neural function score was confirmed after 6h of establishing model, the protein expression levels of ERK5, Caspase-3 and Caspase-9 were detected using RT-PCR and Western blot, and the apoptosis of nerve cells in the cerebral hemorrhage model was detected using TUNEL staining.
Results The neurological function score in experimental group was higher than that in model group, and lower than that in blank control group and sham operation group (P ＜ 0.01). The differences of the relative expression levels of ERK5, Caspase-3 and Caspase-9 mRNA among the blank control group, model group, sham operation group and experimental group were statistically significant (P ＜ 0.05). The relative expression levels of ERK5, Caspase-3 and Caspase-9 mRNA in experimental group were higher than those in blank control group and sham operation group, and lower than those in model group (P ＜ 0.05). The relative expression levels of ERK5, Caspase-3 and Caspase-9 protein in experimental group were lower than those in model group, and higher than those in blank control group and sham operation group (P ＜ 0.01). The results of TUNEL staining showed that the apoptotic index in experimental group was lower than that in model group, and higher than that in sham operation group and blank control group (P ＜ 0.05).
Conclusions The ERK5 signaling pathway may mediate the apoptosis by regulating the expression levels of Caspase-3 and Caspase-9, and the down-regulation of ERK5 protein expression may play protective effects on nerve cells.

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