Isoflurane affects the cytoskeleton but not survival, proliferation, or synaptogenic properties of rat astrocytes in vitro
Isoflurane affects the cytoskeleton but not survival, proliferation, or synaptogenic properties of rat astrocytes in vitro
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DOI：
10.1093/bja/aet169
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发表时间：
2013-06-01
影响因子：
9.8
通讯作者：
Crosby, G.
中科院分区：
文献类型：
--
作者：
Culley, D. J.;Cotran, E. K.;Crosby, G.
大脑中超过一半的细胞是神经胶质细胞，但全身麻醉剂对这些细胞的影响在很大程度上还没有得到研究。我们假设星形胶质细胞与神经元的健康和突触的形成和功能密切相关，容易受到异氟烷的不利影响。培养的大鼠星形胶质细胞在空气或空气中仅用1.4异氟烷处理4h。在处理结束或2天后，通过比色分析、免疫细胞化学或迁移分析来评估细胞的活力、增殖和细胞骨架。此外，原代培养的大鼠皮质神经元在对照组[星形胶质细胞条件培养液(ACM)]或异氟烷暴露的星形胶质细胞(Iso-ACM)的条件培养液中处理4d，用突触素1和PSD-95免疫组织化学染色检测突触点状突触。基于胸苷类似物的掺入，它也没有抑制增殖。异氟醚对F-肌动蛋白无影响，但降低了微管蛋白和胶质纤维酸性蛋白的表达(分别为P<0.05和P<0.01)，但不影响星形胶质细胞的运动能力。异氟醚可减少体外培养的星形胶质细胞微管和中间丝蛋白的表达，但不影响星形胶质细胞的存活率、增殖性、运动性和支持突触的能力。
More than half of the cells in the brain are glia and yet the impact of general anaesthetics on these cells is largely unexamined. We hypothesized that astroglia, which are strongly implicated in neuronal well-being and synapse formation and function, are vulnerable to adverse effects of isoflurane.Cultured rat astrocytes were treated with 1.4 isoflurane in air or air alone for 4 h. Viability, proliferation, and cytoskeleton were assessed by colorimetric assay, immunocytochemistry, or a migration assay at the end of treatment or 2 days later. Also, primary rat cortical neurones were treated for 4 days with conditioned medium from control [astrocyte-conditioned media (ACM)], or isoflurane-exposed astrocytes (Iso-ACM) and synaptic puncta were assessed by synapsin 1 and PSD-95 immunostaining.By several measures, isoflurane did not kill astrocytes. Nor, based on incorporation of a thymidine analogue, did it inhibit proliferation. Isoflurane had no effect on F-actin but reduced expression of -tubulin and glial fibrillary acidic protein both during exposure (P0.05 and P0.001, respectively) and 2 days later (P0.01), but did not impair astrocyte motility. ACM increased formation of PSD-95 but not synapsin 1 positive puncta in neuronal cultures, and Iso-ACM was equally effective.Isoflurane decreased expression of microtubule and intermediate filament proteins in astrocytes in vitro, but did not affect their viability, proliferation, motility, and ability to support synapses.