Zhou Xueying, Hu Ji, Xu Deyi, Zhang Sheng, Wang Qianyan
Department of Psychiatry, Liyuan Hospital of Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430077, P.R. China.
Department of Anesthesiology, Liyuan Hospital of Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430077, P.R. China.
Exp Ther Med. 2023 Feb 10;25(3):134. doi: 10.3892/etm.2023.11833. eCollection 2023 Mar.
Dementia is defined as memory loss and other cognitive decline and it severely influences daily life. Alzheimer's disease (AD) is the most common cause of dementia. Dedicator of cytokinesis 8 (DOCK8) is reported to be involved in neurological diseases. The present study focused on investigating the role that DOCK8 serves in AD and addressing its hidden regulatory mechanism. Initially, Aβ (Aβ) was applied for the administration of BV2 cells. Subsequently, the mRNA and protein expression levels of DOCK8 were evaluated utilizing reverse transcription-quantitative PCR (RT-qPCR) and western blotting. After the DOCK8 silencing, immunofluorescence staining (IF), ELISA, wound healing and Transwell assays were applied to assess ionized calcium binding adapter molecule-1 (IBA-1) expression, release of inflammatory factors, migration and invasion in Aβ-induced BV2 cells. IF was used to evaluate cluster of differentiation (CD)11b expression. RT-qPCR and western blotting were to analyze the levels of M1 cell markers inducible nitric oxide synthase (iNOS) and CD86. The expression of STAT3/NLR family pyrin domain containing 3 (NLRP3)/NF-κB signaling-related proteins were determined by western blotting. Finally, the viability and apoptosis in hippocampal HT22 cells with DOCK8 depletion were estimated. Results revealed that Aβ induction greatly stimulated the expression levels of IBA-1 and DOCK8. DOCK8 silencing suppressed Aβ-induced inflammation, migration and invasion of BV2 cells. Additionally, DOCK8 deficiency conspicuously decreased the expression levels of CD11b, iNOS and CD86. The expression of phosphorylated (p-)STAT3, NLRP3, ASC, caspase1 and p-p65 was downregulated in Aβ-induced BV2 cells after DOCK8 depletion. STAT3 activator Colivelin reversed the effects of DOCK8 knockdown on IBA-1 expression, inflammation, migration, invasion and M1 cell polarization. In addition, the viability and apoptosis in hippocampal HT22 cells stimulated by neuroinflammatory release of BV2 cells were repressed following DOCK8 deletion. Collectively, DOCK8 interference alleviated Aβ-induced damage of BV2 cells by inhibiting STAT3/NLRP3/NF-κB signaling.
痴呆症的定义为记忆丧失及其他认知功能衰退，它严重影响日常生活。阿尔茨海默病（AD）是痴呆症最常见的病因。据报道，细胞分裂素8（DOCK8）参与神经疾病。本研究着重调查DOCK8在AD中所起的作用，并探究其潜在的调控机制。首先，将β淀粉样蛋白（Aβ）应用于BV2细胞给药。随后，利用逆转录定量聚合酶链反应（RT-qPCR）和蛋白质免疫印迹法评估DOCK8的mRNA和蛋白表达水平。在DOCK8沉默后，应用免疫荧光染色（IF）、酶联免疫吸附测定（ELISA）、伤口愈合实验和Transwell实验评估Aβ诱导的BV2细胞中离子钙结合衔接分子1（IBA-1）的表达、炎症因子的释放、迁移和侵袭能力。IF用于评估分化簇（CD）11b的表达。RT-qPCR和蛋白质免疫印迹法用于分析M1细胞标志物诱导型一氧化氮合酶（iNOS）和CD86的水平。通过蛋白质免疫印迹法测定信号转导和转录激活因子3（STAT3）/NOD样受体家族含pyrin结构域蛋白3（NLRP3）/核因子κB（NF-κB）信号相关蛋白的表达。最后，评估DOCK8缺失的海马HT22细胞的活力和凋亡情况。结果显示，Aβ诱导极大地刺激了IBA-1和DOCK8的表达水平。DOCK8沉默抑制了Aβ诱导的BV2细胞炎症、迁移和侵袭。此外，DOCK8缺失显著降低了CD11b、iNOS和CD86的表达水平。在DOCK8缺失后，Aβ诱导的BV2细胞中磷酸化（p-）STAT3、NLRP3、凋亡相关斑点样蛋白（ASC）、半胱天冬酶1和磷酸化p65的表达下调。STAT3激活剂可立维林逆转了DOCK8敲低对IBA-1表达、炎症、迁移、侵袭和M1细胞极化的影响。此外，在DOCK8缺失后，由BV2细胞神经炎症释放刺激的海马HT22细胞的活力和凋亡受到抑制。总的来说，DOCK8干扰通过抑制STAT3/NLRP3/NF-κB信号减轻了Aβ诱导的BV2细胞损伤。
