Department of Anesthesiology and Pain Clinic, The Affiliated Lianyungang Oriental Hospital of Xuzhou Medical University.
Department of Neurology, The Affiliated Lianyungang Oriental Hospital of Xuzhou Medical University.
Drug Des Devel Ther. 2020 Dec 18;14:5589-5598. doi: 10.2147/DDDT.S281582. eCollection 2020.
Neurotoxicity of anesthetics has been widely observed by clinicians. It is reported that inflammation and oxidative stress are involved in the pathological process. In the present study, we aimed to assess the therapeutic effects of agomelatine against isoflurane-induced inflammation and damage to brain endothelial cells.
MTT assay was used to detect cell viability in order to determine the optimized concentration of agomelatine. The bEnd.3 brain endothelial cells were treated with 2% isoflurane in the presence or absence of agomelatine (5, 10 μM) for 24 h. LDH release was evaluated and the ROS levels were checked using DHE staining assay. The expressions of IL-6, IL-8, TNF-α, VEGF, TF, VCAM-1, and ICAM-1 were evaluated using real-time PCR and ELISA. Real-time PCR and Western blot analysis were used to determine the expression level of Egr-1.
The decreased cell viability promoted LDH release and elevated ROS levels induced by isoflurane were significantly reversed by the introduction of agomelatine in a dose-dependent manner. The expression levels of IL-6, IL-8, TNF-α, VEGF, TF, VCAM-1, and ICAM-1 were elevated by stimulation with isoflurane, which were significantly suppressed by the administration of agomelatine. The up-regulation of transcriptional factor Egr-1 induced by isoflurane was down-regulated by agomelatine.
Agomelatine might attenuate isoflurane-induced inflammation and damage via down-regulating Egr-1 in brain endothelial cells.
麻醉药的神经毒性已被临床医生广泛观察到。据报道，炎症和氧化应激参与了病理过程。在本研究中，我们旨在评估阿戈美拉汀对异氟烷诱导的脑内皮细胞炎症和损伤的治疗作用。
MTT 法检测细胞活力，以确定阿戈美拉汀的最佳浓度。用 2%异氟烷处理 bEnd.3 脑内皮细胞，同时加入或不加入阿戈美拉汀（5、10 μM）24 小时。评估 LDH 释放，并用 DHE 染色法检测 ROS 水平。用实时 PCR 和 ELISA 评估 IL-6、IL-8、TNF-α、VEGF、TF、VCAM-1 和 ICAM-1 的表达。用实时 PCR 和 Western blot 分析测定 Egr-1 的表达水平。
异氟烷诱导的细胞活力降低、LDH 释放增加和 ROS 水平升高，均被阿戈美拉汀呈剂量依赖性显著逆转。异氟烷刺激后，IL-6、IL-8、TNF-α、VEGF、TF、VCAM-1 和 ICAM-1 的表达水平升高，而阿戈美拉汀给药则显著抑制了这些表达水平。异氟烷诱导的转录因子 Egr-1 的上调被阿戈美拉汀下调。
阿戈美拉汀可能通过下调脑内皮细胞中的 Egr-1 来减轻异氟烷诱导的炎症和损伤。
