Microglia induce neural cell death via a proximity-dependent mechanism involving nitric oxide
Microglia induce neural cell death via a proximity-dependent mechanism involving nitric oxide
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DOI：
10.1016/j.brainres.2006.02.032
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发表时间：
2006-04-21
期刊：
影响因子：
2.9
通讯作者：
Dragunow, Mike
中科院分区：
文献类型：
--
作者：
Gibbons, Hannah M.;Dragunow, Mike
小胶质细胞通过加剧神经元和非神经元细胞死亡在许多神经系统疾病的发病机制中发挥重要作用，但所涉及的机制尚不清楚。为了研究小胶质细胞-神经元相互作用，我们在共培养系统中使用了小鼠 BV-2 小胶质细胞系和人类神经元样 SK-N-SH 神经母细胞瘤细胞系，该系统能够实现邻近依赖性相互作用和通信，跨孔系统仅允许通过可扩散分子进行邻近独立通信，以及条件培养基系统，通过该系统不可能实现邻近依赖性相互作用或细胞间通信。用脂多糖和干扰素-γ (LPS/IFN-γ) 激活 BV-2 细胞会降低单独生长的 BV-2 细胞以及与 SK-N-SH 细胞共培养的 BV-2 细胞的活力，但不会降低单独生长的 SK-N-SH 细胞的活力。相反，在跨孔和条件培养基系统中激活 BV-2 细胞对 SK-N-SH 细胞的活力没有任何影响，表明小胶质细胞必须靠近神经细胞才能引发细胞毒性。为了确定参与邻近依赖性细胞死亡的分子，研究了小胶质细胞激活的抑制剂。只有特异性诱导型一氧化氮合酶 (iNOS) 抑制剂 S-甲基异硫脲和已知可抑制小胶质细胞 NOS 表达的低温，才能在 LPS/IFN-γ 激活后防止细胞死亡。这些结果表明，激活的小胶质细胞释放一氧化氮，这至少部分是造成邻近依赖性小胶质细胞介导的神经毒性的原因。 (c) 2006 Elsevier B.V. 保留所有权利。
Microglial cells play a major role in the pathogenesis of many neurological diseases by exacerbating neuronal and non-neuronal cell death, but the mechanisms involved are unclear. To investigate the microglial-neuronal interactions, we used the murine BV-2 microglial cell line and the human neuronal-like SK-N-SH neuroblastoma cell line in a co-culture system that enabled proximity-dependent interaction and communication, a trans-well system that allowed proximity-independent communication through diffusible molecules only, and a conditioned media system through which no proximity-dependent interactions or cell-to-cell communication is possible. Activation of BV-2 cells with lipopolysaccharide and interferon-gamma (LPS/IFN-gamma) decreased viability of the BV-2 cells alone and in co-cultures with SK-N-SH cells, but not SK-N-SH cells grown alone. In contrast, activation of BV-2 cells in the trans-well and conditioned media system did not have any effect on the viability of SK-N-SH cells, suggesting that microglia must be in close proximity to the neural cells to elicit cytotoxicity. To determine the molecules involved in proximity-dependent cell death, inhibitors of microglial activation were investigated. Only the specific inducible nitric oxide synthase (iNOS) inhibitor S-methylisothiourea, and hypothermia, which is known to suppress microglial NOS expression, prevented cell death after LPS/IFN-gamma activation. These results suggest that activated microglia release nitric oxide that is, at least partially, responsible for proximity-dependent micro glial-mediated neural toxicity. (c) 2006 Elsevier B.V. All rights reserved.