Retinal ganglion cell line apoptosis induced by hydrostatic pressure
Retinal ganglion cell line apoptosis induced by hydrostatic pressure
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DOI：
10.1016/j.brainres.2006.02.061
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发表时间：
2006-05-01
期刊：
影响因子：
2.9
通讯作者：
Hill, Mark A.
中科院分区：
文献类型：
--
作者：
Agar, Ashish;Li, Shaojuan;Hill, Mark A.
细胞对压力变化的反应与许多疾病过程有关。在青光眼中，视网膜神经节细胞（RGCs）的凋亡与眼内压升高有关，然而，确切的细胞机制仍不清楚。我们以前已经表明，压力可以诱导凋亡的B35和PC 12神经元细胞系，使用体外模型的压力升高。一种新的RGC线使我们能够研究压力对视网膜神经元的影响。在我们的模型中，“RGC-5”培养物经受升高的环境静水压力条件。实验压力条件为100 mm Hg和30 mm Hg，代表急性（高）和慢性（低压）青光眼，正常眼内压为15 mm Hg，设置为高于大气压2小时。除了施加压力之外，阴性对照被相同地处理，而阳性对照通过用已知的凋亡刺激物处理来产生。通过细胞形态学和特异性TUNEL和Annexin V荧光标记物的组合来确定细胞凋亡。同时通过激光扫描细胞仪（LSC）进行评估，这也使得定量标记物分析成为可能。与对照组相比，RGC-5神经元的凋亡细胞比例显著增加;在100 mm Hg时最大，在30 mm Hg时中等，在15 mm Hg时无统计学显著性。该分级反应与压力升高水平成比例，代表类似临床环境（急性、慢性青光眼和正常）的严重程度。这些结果补充了压力诱导的细胞凋亡在其他神经元培养的早期发现。他们提出了压力相关的机械转导和细胞死亡的新机制的可能性，与青光眼等疾病的发病机制有关。(c)2006 Elsevier B. V.保留所有权利。
Cellular responses to changes in pressure are implicated in numerous disease processes. In glaucoma apoptosis of retinal ganglion cells (RGCs) is associated with elevated intra-ocular pressure, however, the exact cellular mechanisms remain unclear. We have previously shown that pressure can induce apoptosis in B35 and PC12 neuronal cell lines, using an in vitro model for pressure elevation. A novel RGC line allows us to study the effects of pressure on retinal neurons. 'RGC-5' cultures were subjected to elevated ambient hydrostatic pressure conditions in our model. Experimental pressure conditions were 100mm Hg and 30 mm Hg, representing acute (high) and chronic (lower-pressure) glaucoma, and 15 mm Hg for normal intra-ocular pressure, set above atmospheric pressure for 2h. Negative controls were treated identically except for the application of pressure, while positive controls were generated by treatment with a known apoptotic stimulus. Apoptosis was determined by a combination of cell morphology and specific TUNEL and Annexin V fluorescent markers. These were assessed simultaneously by laser scanning cytometry (LSC), which also enabled quantitative marker analysis. RGC-5 neurons showed a significantly increased proportion of apoptotic cells compared with controls; maximal at 100mm Hg, moderate at 30mm Hg and not statistically significant at 15mm Hg. This graded response, proportionate to the level of pressure elevation, is representative of the severity of analogous clinical settings (acute, chronic glaucoma and normal). These results complement earlier findings of pressure-induced apoptosis in other neuronal cultures. They suggest the possibility of novel mechanisms of pressure-related mechanotransduction and cell death, relevant to the pathogenesis of diseases such as glaucoma. (c) 2006 Elsevier B.V. All rights reserved.