Connexin43 antisense oligodeoxynucleotide treatment down-regulates the inflammatory response in an in vitro interphase organotypic culture model of optic nerve ischaemia
Connexin43 antisense oligodeoxynucleotide treatment down-regulates the inflammatory response in an in vitro interphase organotypic culture model of optic nerve ischaemia
复制标题
DOI：
10.1016/j.jocn.2008.08.002
复制
发表时间：
2008-11-01
影响因子：
2
通讯作者：
Green, Colin R.
中科院分区：
文献类型：
--
作者：
Danesh-Meyer, Helen V.;Huang, Rex;Green, Colin R.
本研究使用视神经缺血模型，研究了体外培养的离体大鼠视神经节段的氧糖剥夺（OGD）。此后，在同一模型中评估了间隙连接蛋白 connexin43 (Cx43) 特异的反义寡脱氧核苷酸 (ASODN) 的作用。暴露于 OGD 2 小时后，在暴露或不暴露于 Cx43 ASODN 的情况下，将视神经维持在间期器官型培养物中。在培养后2小时、6小时以及第1、2、3和6天对视神经进行切片。使用碘化丙啶 (PI) 染色和 Cx43、毛细血管（血管性血友病因子）、星形胶质细胞（胶质纤维酸性蛋白）、小胶质细胞和内皮细胞（异凝集素 B4）的特异性标记物对细胞死亡进行定量，并结合数字光学和共聚焦显微镜评估这些参数。在该模型中，视神经暴露于 OGD 后 2 小时观察到 Cx43 上调，并在第 3 天达到峰值。Cx43 ASODN 治疗抑制了这种上调。此外，在对照视神经节段的中心发现了比治疗神经更多的 PI 标记细胞 (p < 0.01)。与 Cx43 ASODN 处理的神经相比，对照还显示出毛细血管破裂的证据以及星形胶质细胞和活化小胶质细胞数量的增加 (p
Using a model of optic nerve ischaemia, this study investigated oxygen-glucose deprivation (OGD) on isolated rat optic nerve segments cultured in vitro. Thereafter, the effect of antisense oligodeoxynucleotides (ASODN) specific to the gap junction protein connexin43 (Cx43) was evaluated in this same model. Following exposure to OGD for 2 hours, optic nerves were maintained in interphase organotypic culture with and without exposure to Cx43 ASODN. Optic nerves were sectioned at 2 hours, 6 hours, and at days 1, 2, 3 and 6 following culture. Cell death was quantified using propidium iodide (PI) staining and specific markers for Cx43, capillaries (von Willebrand factor) astrocytes (glial fibrillary acidic protein), microglia and endothelial cells (isolectin B4) were used to evaluate these parameters in conjunction with digital light and confocal microscopy. In this model, up-regulation of Cx43 was seen at 2 hours following exposure of the optic nerve to OGD and peaked at day 3. Cx43 ASODN treatmetn dampened this up-regulation. Additionally, more PI labeld cells were found in the centre of control optic nerve segments than in treated nerves (p < 0.01). Controls also showed evidence of capillary breakdown and increased numbers of astrocytes and activated microglia compared to Cx43 ASODN treated nerves (p