目的:我们先前的研究发现,氧糖剥夺/再灌注(OGD/R)后,ErbB4基因表达发生变化。然而,ErbB4在脑缺血中的确切作用和机制尚不清楚。在这项研究中,我们探讨了ErbB4在OGD/R后的保护作用及其可能的机制。
方法:体外和体内建立脑缺血/再灌注(I/R)损伤模型。细胞活力,凋亡,通过MTT测量ROS的产生,TUNEL,和荧光探针2',7'-二氯荧光素二乙酸酯(DCFH-DA)。通过TTC评估梗死面积。我们进行了生物信息学分析,以筛选涉及ErbB4变化的新关键基因。RNA-Seq用于转录组分析。通过定量RT-PCR和蛋白质印迹检测RNA和蛋白质表达。
结果:在体外和体内脑I/R损伤后,80-kDaErbB4的表达降低。共表达网络分析显示ErbB4的表达与Adrb1、Adrb2、Ldlr、Dab2定量RT-PCR显示,Adrb1,Adrb2和Dab2的mRNA表达水平上调,OGD/R后Ldlr下降神经调节素1(NRG1)激活ErbB4表达显著促进细胞存活,减弱海马细胞凋亡,OGD/R后ROS产量降低此外,使用特异性siRNA消除ErbB4逆转了这些有益作用.
结论:我们的数据揭示了ErbB4对OGD/R损伤的神经保护作用,并且该作用可能与ErbB4膜相关片段的变化以及Adrb1,Adrb2,Ldlr,Dab2
OBJECTIVE: Our previous study found that
ErbB4 gene expression was changed after oxygen-glucose deprivation/reperfusion (OGD/R). However, the exact role and mechanism of
ErbB4 in brain ischemia are largely unknown. In this study, we explored the protective effects of
ErbB4 and its possible mechanism after OGD/R.
METHODS: Cerebral ischemia/reperfusion (I/R) injury model was established in vitro and in vivo. Cell viability, apoptosis, and ROS production were measured by MTT, TUNEL, and fluorescent probe 2\', 7\'-dichlorofluorescein diacetate (DCFH-DA). Infarct size was evaluated by TTC. We performed bioinformatics analyses to screen for novel key genes involved in
ErbB4 changes. RNA-Seq was used to transcriptome analysis. RNA and protein expression were detected by quantitative RT‒PCR and western bloting.
RESULTS: The expression of 80-kDa ErbB4 decreased after cerebral I/R injury in vitro and in vivo. Co-expression network analysis revealed that ErbB4 expression was correlated with the changes in Adrb1, Adrb2, Ldlr, and Dab2. Quantitative RT‒PCR revealed that the mRNA expression levels of Adrb1, Adrb2, and Dab2 were upregulated, and that of Ldlr was decreased after OGD/R. Activation of ErbB4 expression by neuregulin 1 (NRG1) significantly promoted cell survival, attenuated hippocampal apoptosis, and decreased ROS production after OGD/R. Furthermore, the elimination of
ErbB4 using a specific siRNA reversed these beneficial effects.
CONCLUSIONS: Our data revealed the neuroprotective effects of ErbB4 against OGD/R injury, and the action could be related to changes in the ErbB4 membrane-associated fragment and the expression of Adrb1, Adrb2, Ldlr, and Dab2.