Multiplexed cytokine protein expression profiles from spreading depression in hippocampal organotypic cultures
Multiplexed cytokine protein expression profiles from spreading depression in hippocampal organotypic cultures
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DOI：
10.1097/01.wcb.0000126566.34753.30
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发表时间：
2004-08-01
影响因子：
6.3
通讯作者：
Kraig, TP
中科院分区：
文献类型：
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作者：
Kunkler, PE;Hulse, RE;Kraig, TP
细胞因子参与缺血耐受，包括由扩散性抑制(SD)引发的耐受，但它们在神经保护中的作用尚不完全清楚。后者可能源于这些信号分子的多效性，其相互作用的复杂性可能通过同时测量多个靶蛋白来更好地破译。因此，作者使用基于微球的流式细胞仪免疫分析和海马体器官培养(HOTCs)来表征细胞因子(IL[IL]1α、IL-1βIL-2、IL-4、IL-6、IL-10、粒细胞-巨噬细胞集落刺激因子[GM-CSF]、干扰素-γ[干扰素-γ]和肿瘤坏死因子-α[TNF-α])对S-GM-CSF的反应的大小、时间过程和多样性。然而，SD在6小时后在HOTCs中引发了7种细胞因子的显著、普遍的增加，剩余的细胞因子IL-10在第1天和第3天变得明显不同。此外，这些变化扩展到包括周围的IL-6和肿瘤坏死因子-α的培养上清液1天和3天。通过IL-1α、IL-1β和干扰素-γ的免疫染色，这种增加局限于小胶质细胞。IL-10虽然在SD后6小时在HOTC中含量显著增加，但在那个时候和第一天在周围介质中的含量明显减少。最后，组织细胞因子的普遍早期升高在恢复3天后稳定下来，以IL-1α、IL-1β和肿瘤坏死因子-α的变化为中心，这些细胞因子能够调节缺血性损伤。
Cytokines are involved in ischemic tolerance, including that triggered by spreading depression (SD), yet their roles in neuroprotection remain incompletely defined. The latter may stem from the pleiotropic nature of these signaling molecules whose complexities for interaction might be better deciphered through simultaneous measurement of multiple targeted proteins. Accordingly, the authors used microsphere-based flow cytometric immunoassays and hippocampal organotypic Cultures (HOTCs) to characterize the magnitude, time course, and diversity of cytokine (interleukin [IL] 1alpha, IL-1beta IL-2, IL-4, IL-6, IL-10, granulocyte-macrophage colony-stimulating factor [GM-CSF], interferon-gamma [IFN-gamma], and tumor necrosis factor-alpha [TNF-alpha]) response to S. GM-CSF was not detected in HOTCs or media. However, SD triggered a significant, generalized increase in seven cytokines evident in HOTCs 6 hours later, with the remaining cytokine, IL-10, becoming significantly different at 1 and 3 days. Additionally, these changes extended to include surrounding media for IL-6 and TNF-alpha by 1 and 3 days. This increase was localized to microglia via immunostaining for IL-1alpha, IL-1beta, and interferon-gamma. IL-10 although significantly more abundant in HOTCs 6 hours after SD, was significantly less abundant in surrounding media at that time and at I day. Finally, the generalized early increase in tissue cytokines later settled to a pattern at 3 days of recovery centering on changes in IL-1alpha, IL-1beta, and TNF-alpha, cytokines capable of modulating ischemic injury.