Department of Neurology, Medical Faculty, RWTH Aachen University, 52074 Aachen, Germany.
Institute of Biochemistry and Molecular Immunology, Medical Faculty, RWTH Aachen University, 52074 Aachen, Germany.
Int J Mol Sci. 2020 Jul 7;21(13):4795. doi: 10.3390/ijms21134795.
Acute ischemic stroke (AIS) is a devastating neurological condition with a lack of neuroprotective therapeutic options, despite the reperfusion modalities thrombolysis and thrombectomy. Post-ischemic brain damage is aggravated by an excessive inflammatory cascade involving the activation and regulation of the pro-inflammatory cytokines IL-1β and IL-18 by inflammasomes. However, the role of AIM2 and NLRC4 inflammasomes and the influence of the neuroprotective steroids 17β-estradiol (E2) and progesterone (P) on their regulation after ischemic stroke have not yet been conclusively elucidated. To address the latter, we subjected a total of 65 rats to 1 h of transient Middle Cerebral Artery occlusion (tMCAO) followed by a reperfusion period of 72 h. Moreover, we evaluated the expression and regulation of AIM2 and NLRC4 in glial single-cell cultures (astroglia and microglia) after oxygen-glucose deprivation (OGD). The administration of E2 and P decreased both infarct sizes and neurological impairments after cerebral ischemia in rats. We detected a time-dependent elevation of gene and protein levels (Western Blot/immunohistochemistry) of the AIM2 and NLRC4 inflammasomes in the post-ischemic brains. E2 or P selectively mitigated the stroke-induced increase of AIM2 and NLRC4. While both inflammasomes seemed to be exclusively abundant in neurons under physiological and ischemic conditions in vivo, single-cell cultures of cortical astrocytes and microglia equally expressed both inflammasomes. In line with the in vivo data, E and P selectively reduced AIM2 and NLRC4 in primary cortical astrocytes and microglial cells after OGD. In conclusion, the post-ischemic elevation of AIM2 and NLRC4 and their down-regulation by E2 and P may shed more light on the anti-inflammatory effects of both gonadal hormones after stroke.
急性缺血性脑卒中(AIS)是一种破坏性的神经疾病,尽管有再灌注方式溶栓和血栓切除术,但缺乏神经保护治疗选择。缺血后脑损伤加剧,原因是炎症小体激活和调节促炎细胞因子 IL-1β和 IL-18,导致过度炎症级联反应。然而,AIM2 和 NLRC4 炎症小体的作用以及神经保护类固醇 17β-雌二醇(E2)和孕酮(P)对它们在缺血性中风后的调节作用尚未得到明确阐明。为了解决后者,我们总共将 65 只大鼠置于 1 小时的短暂性大脑中动脉闭塞(tMCAO)后,进行 72 小时的再灌注期。此外,我们评估了在氧葡萄糖剥夺(OGD)后神经胶质单细胞培养物(星形胶质细胞和小胶质细胞)中 AIM2 和 NLRC4 的表达和调节。E2 和 P 的给药可减少大鼠脑缺血后的梗死面积和神经损伤。我们检测到缺血后大脑中 AIM2 和 NLRC4 炎症小体的基因和蛋白水平(Western Blot/免疫组织化学)呈时间依赖性升高。E2 或 P 选择性减轻了中风引起的 AIM2 和 NLRC4 增加。虽然两种炎症小体在体内生理和缺血条件下似乎仅在神经元中丰富,但皮质星形胶质细胞和小胶质细胞的单细胞培养物同样表达两种炎症小体。与体内数据一致,E 和 P 选择性降低了 OGD 后原代皮质星形胶质细胞和小胶质细胞中的 AIM2 和 NLRC4。总之,缺血后 AIM2 和 NLRC4 的升高及其被 E2 和 P 下调可能会进一步阐明两种性腺激素在中风后的抗炎作用。