Institute of Biomedical Sciences, National Chung-Hsing University, Taichung 40227, Taiwan.
Department of Medical Research, Taichung Veterans General Hospital, Taichung 40210, Taiwan.
Int J Mol Sci. 2023 Jul 13;24(14):11416. doi: 10.3390/ijms241411416.
Thrombin is a multifunctional serine protein which is closely related to neurodegenerative disorders. The Aryl hydrocarbon receptor (AhR) is well expressed in microglia cells involving inflammatory disorders of the brain. However, it remains unclear as to how modulation of AhR expression by thrombin is related to the development of neurodegeneration disorders. In this study, we investigated the role of AhR in the development of thrombin-induced neurodegenerative processes, especially those concerning microglia. The primary culture of either wild type or AhR deleted microglia, as well as BV-2 cell lines, was used for an in vitro study. Hippocampal slice culture and animals with either wild type or with AhR deleted were used for the ex vivo and in vivo studies. Simulations of ligand protein docking showed a strong integration between the thrombin and AhR. In thrombin-triggered microglia cells, deleting AhR escalated both the NO release and iNOS expression. Such effects were abolished by the administration of the AhR agonist. In thrombin-activated microglia cells, downregulating AhR increased the following: vascular permeability, pro-inflammatory genetic expression, MMP-9 activity, and the ratio of M1/M2 phenotype. In the in vivo study, thrombin induced the activation of microglia and their volume, thereby contributing to the deterioration of neurobehavior. Deleting AhR furthermore aggravated the response in terms of impaired neurobehavior, increasing brain edema, aggregating microglia, and increasing neuronal death. In conclusion, thrombin caused the activation of microglia through increased vessel permeability, expression of inflammatory response, and phenotype of M1 microglia, as well the MMP activity. Deleting AhR augmented the above detrimental effects. These findings indicate that the modulation of AhR is essential for the regulation of thrombin-induced brain damages and that the AhR agonist may harbor the potentially therapeutic effect in thrombin-induced neurodegenerative disorder.
凝血酶是一种多功能丝氨酸蛋白酶,与神经退行性疾病密切相关。芳香烃受体(AhR)在小胶质细胞中表达丰富,涉及大脑炎症性疾病。然而,凝血酶对 AhR 表达的调节与神经退行性疾病的发展之间的关系尚不清楚。在这项研究中,我们研究了 AhR 在凝血酶诱导的神经退行性过程中的作用,特别是涉及小胶质细胞的过程。使用野生型或 AhR 缺失的小胶质细胞原代培养物以及 BV-2 细胞系进行体外研究。使用海马切片培养物和野生型或 AhR 缺失的动物进行离体和体内研究。配体蛋白对接模拟显示凝血酶和 AhR 之间存在很强的整合。在凝血酶触发的小胶质细胞中,缺失 AhR 会增加 NO 释放和 iNOS 表达。这些作用被 AhR 激动剂的给药所消除。在凝血酶激活的小胶质细胞中,下调 AhR 会增加血管通透性、促炎基因表达、MMP-9 活性和 M1/M2 表型的比值。在体内研究中,凝血酶诱导小胶质细胞的激活及其体积增加,从而导致神经行为恶化。进一步缺失 AhR 会加重神经行为受损、脑水肿增加、小胶质细胞聚集和神经元死亡的反应。总之,凝血酶通过增加血管通透性、炎症反应表达和 M1 小胶质细胞表型以及 MMP 活性来激活小胶质细胞。缺失 AhR 增强了上述有害作用。这些发现表明,AhR 的调节对于调节凝血酶诱导的脑损伤是必要的,AhR 激动剂可能在凝血酶诱导的神经退行性疾病中具有潜在的治疗作用。