Interleukin-10 receptor signaling through STAT-3 regulates the apoptosis of retinal ganglion cells in response to stress
Interleukin-10 receptor signaling through STAT-3 regulates the apoptosis of retinal ganglion cells in response to stress
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DOI：
10.1167/iovs.03-0534
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发表时间：
2003-12-01
影响因子：
4.4
通讯作者：
Patil, RV
中科院分区：
文献类型：
--
作者：
Boyd, ZS;Kriatchko, A;Patil, RV
目的.白细胞介素（IL）-10最近已被证明可以促进神经元和神经胶质细胞的存活。本报告的目的是研究IL-10是否有任何作用，在保护视网膜神经节细胞（RGC）死亡的条件下，生长因子被删除，或在氧化应激存在。在两种应激条件下研究了激活RGCs中IL-10信号传导的信号转导途径。测定各种白细胞介素对RGC细胞系活力的影响，并定量凋亡细胞。进行免疫印迹分析以鉴定RGC提取物中的IL-10受体（IL-10 R）和磷酸化或非磷酸化Akt和STAT-3蛋白。用免疫组织化学方法对大鼠视网膜切片进行鉴定。96小时后，在生长因子存在或不存在地塞米松（1 μ M）的情况下，分别有68.5% +/- 3.4%和53.4% +/- 2.6%的细胞发生RGCs凋亡。添加浓度为50 ng/mL的IL-10显著降低了RGCs的凋亡群体，在不存在生长因子和地塞米松的情况下降低至28.2% +/-2.3%，在不存在单独生长因子的情况下降低至31% +/-2.7%。RGC以及天然视网膜表达功能性IL-10 R，如通过免疫印迹分析和通过IL-10磷酸化Stat-3的能力所确定的。然而，IL-10不能磷酸化Akt。IL-10导致59%和42%的血清剥夺细胞的细胞凋亡群体与地塞米松治疗，分别减少。这些观察结果证实，IL-10 R的活化促进了RGC的存活，并且这种存活促进活性是由于IL-10通过Stat-3途径而不是通过Akt细胞存活途径进行信号传导，所述Stat-3途径抑制细胞死亡。
PURPOSE. Interleukin (IL)-10 has recently been shown to promote survival of neurons and glia. The purpose of this report is to investigate whether IL-10 has any role in protecting retinal ganglion cells (RGCs) from death under conditions in which growth factors are removed, or in which oxidative stress is present. Signal transduction pathways that activate IL-10 signaling in RGCs were studied in both stress conditions.METHODS. Effects of various interleukins on the viability of the RGC cell line was determined, and apoptotic cells were quantified. Immunoblot analysis was preformed to identify the IL-10 receptor (IL-10R) and phosphorylated or nonphosphorylated Akt and STAT-3 proteins in RGC extracts. Immunohistochemistry was performed on the rat retinal sections to identify native IL-10R.RESULTS. Apoptosis of RGCs in the absence of growth factors with or without dexamethasone (1 muM) occurred in 68.5% +/- 3.4% and 53.4% +/- 2.6% of cells, respectively, after 96 hours. Addition of IL-10 at a concentration of 50 ng/mL significantly reduced the apoptotic population of RGCs to 28.2% +/- 2.3% in the absence of growth factors with dexamethasone and to 31% +/- 2.7% in the absence of growth factors alone. RGCs as well as native retina expressed functional IL-10R as determined by immunoblot analysis and by the ability of IL-10 to phosphorylate Stat-3. However, IL-10 failed to phosphorylate Akt in these cells.CONCLUSIONS. IL-10 caused a 59% and 42% reduction in the apoptotic population of serum-deprived cells with and without dexamethasone treatment, respectively. These observations establish that activation of IL-10R promotes survival of RGCs and this survival-promoting activity is due to IL-10 signaling through the Stat-3 pathway, which inhibits the cell death and not through the Akt cell survival pathway.