New Therapeutic Targets Group (TargetsLab), Departament de Ciències Mèdiques, Facultat de Medicina, Universitat de Girona, E-17071 Girona, Spain.
Grifols Bioscience Research Group, Barcelona, Spain.
Biochim Biophys Acta Gen Subj. 2018 Sep;1862(9):1852-1861. doi: 10.1016/j.bbagen.2018.05.017. Epub 2018 May 30.
Death due to cerebral stroke afflicts a large number of neuronal populations, including glial cells depending on the brain region affected. Drugs with a wide cellular range of protection are needed to develop effective therapies for stroke. Human alpha 1-antitrypsin (hAAT) is a serine proteinase inhibitor with potent anti-inflammatory, anti-apoptotic and immunoregulatory activities. This study aimed to test whether hAAT can protect different kind of neurons and glial cells after the oxygen and glucose deprivation (OGD).
Addition of hAAT to mouse neuronal cortical, hippocampal and striatal cultures, as well as glial cultures, was performed 30 min after OGD induction and cell viability was assessed 24 h later. The expression of different apoptotic markers and several inflammatory parameters were assessed by immunoblotting and RT-PCR.
hAAT had a concentration-dependent survival effect in all neuronal cultures exposed to OGD, with a maximal effect at 1-2 mg/mL. The addition of hAAT at 1 mg/mL reduced the OGD-mediated necrotic and apoptotic death in all neuronal cultures. This neuroprotective activity of hAAT was associated with a decrease of cleaved caspase-3 and an increase of MAP2 levels. It was also associated with a reduction of pro-inflammatory cytokines protein levels and expression, increase of IL-10 protein levels and decrease of nuclear localization of nuclear factor-kappaB. Similar to neurons, addition of hAAT protected astrocytes and oligodendrocytes against OGD-induced cell death.
Human AAT protects neuronal and glial cells against OGD through interaction with cytokines.
Human AAT could be a good therapeutic neuroprotective candidate to treat ischemic stroke.
由于脑中风而导致的死亡会影响大量神经元群体，包括受影响的脑区的神经胶质细胞。需要开发具有广泛细胞保护范围的药物，以针对中风开发有效的治疗方法。人α 1-抗胰蛋白酶（hAAT）是一种丝氨酸蛋白酶抑制剂，具有强大的抗炎、抗凋亡和免疫调节活性。本研究旨在测试 hAAT 是否能在氧葡萄糖剥夺（OGD）后保护不同类型的神经元和神经胶质细胞。
在诱导 OGD 后 30 分钟向小鼠神经元皮质、海马和纹状体培养物以及神经胶质培养物中添加 hAAT，并在 24 小时后评估细胞活力。通过免疫印迹和 RT-PCR 评估不同凋亡标志物和几种炎症参数的表达。
hAAT 在所有暴露于 OGD 的神经元培养物中均具有浓度依赖性的生存效应，在 1-2mg/mL 时达到最大效应。在 1mg/mL 时添加 hAAT 可减少所有神经元培养物中的 OGD 介导的坏死和凋亡性死亡。hAAT 的这种神经保护活性与 cleaved caspase-3 的减少和 MAP2 水平的增加有关。它还与促炎细胞因子蛋白水平的降低、IL-10 蛋白水平的增加和核因子-kappaB 的核定位减少有关。与神经元类似，添加 hAAT 可保护星形胶质细胞和少突胶质细胞免受 OGD 诱导的细胞死亡。
人 AAT 通过与细胞因子相互作用保护神经元和神经胶质细胞免受 OGD 损伤。
人 AAT 可能是治疗缺血性中风的一种良好的治疗性神经保护候选药物。
