Department of Neurology, The University of Chicago Medical Center, Chicago, Illinois 60637, USA.
J Neurochem. 2011 Apr;117(2):187-96. doi: 10.1111/j.1471-4159.2010.07103.x. Epub 2010 Dec 1.
Cold pre-conditioning reduces subsequent brain injury in small animals but the underlying mechanisms remain undefined. As hypothermia triggers systemic macrophage tumor necrosis factor alpha (TNF-α) production and other neural pre-conditioning stimuli depend on this cytokine, we reasoned that microglia and TNF-α would be similarly involved with cold pre-conditioning neuroprotection. Also, as slice cultures closely approximate their in vivo counterpart and include quiescent microglia, we used rat hippocampal slice cultures to confirm this hypothesis. Furthermore, inflammatory cytokine gene screening with subsequent PCR and immunostaining confirmation of targeted mRNA and related protein changes showed that cold pre-conditioning triggered a significant rise in TNF-α that localized to microglia and a significant rise in interleukin (IL)-11 that localized mainly to hippocampal pyramidal neurons and, more rarely, astrocytes. Importantly, co-stimulation with cold and IL-11, an anti-inflammatory cytokine that inhibits TNF-α expression, abrogated the otherwise evident protection. Instead, cold pre-conditioning coupled with blockade of IL-11 signaling further enhanced neuroprotection from that seen with cold pre-conditioning alone. Thus, physiological activation of brain pro-inflammatory cytokine signaling, and its amplification by inhibition of coincident anti-inflammatory cytokine signaling, may be opportune targets for the development of novel therapeutics that can mimic the protection seen in cold pre-conditioning.
冷预处理可减少小动物的后续脑损伤,但潜在机制尚不清楚。由于低温会触发全身巨噬细胞肿瘤坏死因子-α(TNF-α)的产生,而其他神经预处理刺激因素依赖于这种细胞因子,因此我们推断小胶质细胞和 TNF-α也会与冷预处理的神经保护作用有关。此外,由于切片培养与体内情况非常相似且包括静止的小胶质细胞,因此我们使用大鼠海马切片培养物来验证这一假说。此外,通过随后的 PCR 进行炎性细胞因子基因筛选,并对靶向 mRNA 和相关蛋白变化进行免疫染色确认,表明冷预处理会引发 TNF-α显著升高,其定位在小胶质细胞中;白细胞介素(IL)-11 显著升高,主要定位于海马锥体神经元,且罕见地定位于星形胶质细胞。重要的是,冷刺激和 IL-11 的共同刺激(IL-11 是一种抑制 TNF-α表达的抗炎细胞因子)消除了原本明显的保护作用。相反,冷预处理与阻断 IL-11 信号通路的结合进一步增强了冷预处理本身带来的神经保护作用。因此,大脑促炎细胞因子信号的生理激活,以及通过抑制伴随的抗炎细胞因子信号对其进行放大,可能是开发新疗法的恰当靶点,这些疗法可以模拟冷预处理带来的保护作用。