Hydrostatic pressure does not cause detectable changes in survival of human retinal ganglion cells.
Hydrostatic pressure does not cause detectable changes in survival of human retinal ganglion cells.
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静液压不会引起人类视网膜神经节细胞存活的可检测到的变化。
DOI：
10.1371/journal.pone.0115591
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Sanderson J
中科院分区：
文献类型：
--
作者：
Osborne A;Aldarwesh A;Rhodes JD;Broadway DC;Everitt C;Sanderson J
眼压升高是青光眼的主要危险因素。IOP升高的一个后果是眼组织承受的静水压力（HP）增加。研究了HP升高对应激通路信号和视网膜神经节细胞（RGC）存活的影响。设计了一个腔室，使细胞暴露于增加的HP（恒定和波动）。使用质量流量控制器实现精确的压力控制（10-100mmHg）。在无血清的DMEM/HamF12中培养来自供体眼（死后<24小时）的人器官型视网膜培养物（horc）。HP升高与模拟缺血（氧葡萄糖剥夺，OGD）比较。LDH和TUNEL检测细胞死亡和凋亡，qRT-PCR检测RGC标志物表达（THY-1），免疫组化检测RGC数量（NeuN）。Western blot检测活化的p38和JNK。与对照组相比，将HORCs暴露于恒定（60mmHg）或波动（10-100mmHg； 1循环/分钟）压力下24或48小时，不会导致结构完整性丧失、LDH释放、RGC标记物表达（THY-1）减少或RGC丧失。此外，在任何时间点，tunel阳性的neun标记细胞均未增加，表明RGCs的凋亡未增加。OGD增加细胞凋亡，降低RGC标志物表达和RGC数量，并导致24h LDH释放升高。在波动压力（10-100mmHg； 1循环/分钟）持续15、30、60和90分钟的HORCs中，p38和JNK的磷酸化保持不变，而OGD （3h）增加了p38和JNK的激活，并在OGD后90分钟保持升高。直接应用HP对HORCs的RGC存活和应激信号没有可检测到的影响。然而，模拟缺血激活应激通路并导致RGC死亡。这些结果表明，直接HP不会导致离体人视网膜RGCs变性。
Elevated intraocular pressure (IOP) is a major risk factor for glaucoma. One consequence of raised IOP is that ocular tissues are subjected to increased hydrostatic pressure (HP). The effect of raised HP on stress pathway signaling and retinal ganglion cell (RGC) survival in the human retina was investigated. A chamber was designed to expose cells to increased HP (constant and fluctuating). Accurate pressure control (10-100mmHg) was achieved using mass flow controllers. Human organotypic retinal cultures (HORCs) from donor eyes (<24h post mortem) were cultured in serum-free DMEM/HamF12. Increased HP was compared to simulated ischemia (oxygen glucose deprivation, OGD). Cell death and apoptosis were measured by LDH and TUNEL assays, RGC marker expression by qRT-PCR (THY-1) and RGC number by immunohistochemistry (NeuN). Activated p38 and JNK were detected by Western blot. Exposure of HORCs to constant (60mmHg) or fluctuating (10-100mmHg; 1 cycle/min) pressure for 24 or 48h caused no loss of structural integrity, LDH release, decrease in RGC marker expression (THY-1) or loss of RGCs compared with controls. In addition, there was no increase in TUNEL-positive NeuN-labelled cells at either time-point indicating no increase in apoptosis of RGCs. OGD increased apoptosis, reduced RGC marker expression and RGC number and caused elevated LDH release at 24h. p38 and JNK phosphorylation remained unchanged in HORCs exposed to fluctuating pressure (10-100mmHg; 1 cycle/min) for 15, 30, 60 and 90min durations, whereas OGD (3h) increased activation of p38 and JNK, remaining elevated for 90min post-OGD. Directly applied HP had no detectable impact on RGC survival and stress-signalling in HORCs. Simulated ischemia, however, activated stress pathways and caused RGC death. These results show that direct HP does not cause degeneration of RGCs in the ex vivo human retina.
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