Dpt. Biología Celular, Instituto Biomedicina. Universidad de León, Spain.
Dpt. Cirugía Medicina y Anatomia Veterinaria. Universidad de León, Spain.
Biochem Pharmacol. 2018 May;151:26-37. doi: 10.1016/j.bcp.2018.02.029. Epub 2018 Feb 27.
Blood reperfusion of the ischemic tissue after stroke promotes increases in the inflammatory response as well as accumulation of unfolded/misfolded proteins in the cell, leading to endoplasmic reticulum (ER) stress. Both Inflammation and ER stress are critical processes in the delayed death of the cells damaged after ischemia. The aim of this study is to check the putative synergic neuroprotective effect by combining anti-inflammatory and anti-ER stress agents after ischemia.
The study was performed on a two-vessel occlusion global cerebral ischemia model. Animals were treated with salubrinal one hour after ischemia and with robenacoxib at 8 h and 32 h after ischemia. Parameters related to the integrity of the blood-brain barrier (BBB), such as matrix metalloproteinase 9 and different cell adhesion molecules (CAMs), were analyzed by qPCR at 24 h and 48 h after ischemia. Microglia and cell components of the neurovascular unit, including neurons, endothelial cells and astrocytes, were analyzed by immunofluorescence after 48 h and seven days of reperfusion.
Pharmacologic control of ER stress by salubrinal treatment after ischemia, revealed a neuroprotective effect over neurons that reduces the transcription of molecules involved in the impairment of the BBB. Robenacoxib treatment stepped neuronal demise forward, revealing a detrimental effect of this anti-inflammatory agent. Combined treatment with robenacoxib and salubrinal after ischemia prevented neuronal loss and changes in components of the neurovascular unit and microglia observed when animals were treated only with robenacoxib.
Combined treatment with anti-ER stress and anti-inflammatory agents is able to provide enhanced neuroprotective effects reducing glial activation, which opens new avenues in therapies against stroke.
中风后缺血组织的血液再灌注会促进炎症反应的增加以及细胞内未折叠/错误折叠蛋白的积累，导致内质网（ER）应激。炎症和 ER 应激都是缺血后细胞迟发性死亡的关键过程。本研究旨在检查抗炎症和抗 ER 应激药物联合使用后对缺血后的潜在协同神经保护作用。
本研究采用两血管闭塞全脑缺血模型。动物在缺血后 1 小时用 salubrinal 治疗，在缺血后 8 小时和 32 小时用 robenacoxib 治疗。用 qPCR 分析缺血后 24 小时和 48 小时与血脑屏障（BBB）完整性相关的参数，如基质金属蛋白酶 9 和不同的细胞黏附分子（CAMs）。缺血后 48 小时和 7 天再灌注后用免疫荧光法分析小胶质细胞和神经血管单元的细胞成分，包括神经元、内皮细胞和星形胶质细胞。
缺血后用 salubrinal 治疗控制 ER 应激显示出对神经元的神经保护作用，降低了参与 BBB 损伤的分子的转录。Robenacoxib 治疗加速了神经元的死亡，显示出这种抗炎药物的有害作用。缺血后用 robenacoxib 和 salubrinal 联合治疗可防止仅用 robenacoxib 治疗时观察到的神经元丢失以及神经血管单元和小胶质细胞成分的变化。
抗 ER 应激和抗炎药物联合治疗能够提供增强的神经保护作用，减少神经胶质细胞的激活，为中风治疗开辟了新途径。
