Critical Neuroprotective Roles of Heme Oxygenase-1 Induction Against Axonal Injury-Induced Retinal Ganglion Cell Death
Critical Neuroprotective Roles of Heme Oxygenase-1 Induction Against Axonal Injury-Induced Retinal Ganglion Cell Death
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DOI：
10.1002/jnr.23398
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发表时间：
2014-09-01
影响因子：
4.2
通讯作者：
Nakazawa, Toru
中科院分区：
文献类型：
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作者：
Himori, Noriko;Maruyama, Kazuichi;Nakazawa, Toru
虽然轴突损伤诱导显著的视网膜神经节细胞（RGC）死亡，但少数RGC能够在视神经挤压（NC）损伤后存活长达7天。为了开发新的治疗方法，我们着手确定轴突损伤抗性RGC基因表达的变化模式。为了补偿视网膜中RGCs的低密度，我们在成年小鼠中用4Di-10ASP对这些细胞进行逆行标记，并且在NC用荧光激活细胞分选纯化RGCs后7天。用基因芯片测定细胞中的基因表达，用定量PCR（qPCR）测定Ho-1的表达。用免疫组化和免疫印迹法评估蛋白质表达的变化。此外，荧光金标记的RGC的密度在用CoPP（一种有效的HO-1诱导剂）预处理的小鼠的视网膜中计数。微阵列和qPCR分析显示NC后RGC中Ho-1的表达增加。免疫组化显示HO-1阳性细胞出现在神经节细胞层（GCL），细胞计数显示HO-1阳性细胞在GCL中的比例在NC后显著升高。NC后7天，CoPP处理小鼠中的RGC数量显著高于对照小鼠。联合HO-1抑制剂SnPP预处理可抑制CoPP的神经保护作用。这些结果反映了HO-1活性对RGC的变化，这是RGC存活的关键部分。因此，上调HO-1信号可能是青光眼的一种新的治疗策略。(C)2014 Wiley Periodicals，Inc.
Although axonal damage induces significant retinal ganglion cell (RGC) death, small numbers of RGCs are able to survive up to 7 days after optic nerve crush (NC) injury. To develop new treatments, we set out to identify patterns of change in the gene expression of axonal damage-resistant RGCs. To compensate for the low density of RGCs in the retina, we performed retrograde labeling of these cells with 4Di-10ASP in adult mice and 7 days after NC purified the RGCs with fluorescence-activated cell sorting. Gene expression in the cells was determined with a microarray, and the expression of Ho-1 was determined with quantitative PCR (qPCR). Changes in protein expression were assessed with immunohistochemistry and immunoblotting. Additionally, the density of Fluoro-gold-labeled RGCs was counted in retinas from mice pretreated with CoPP, a potent HO-1 inducer. The microarray and qPCR analyses showed increased expression of Ho-1 in the post-NC RGCs. Immunohistochemistry also showed that HO-1-positive cells were present in the ganglion cell layer (GCL), and cell counting showed that the proportion of HO-1-positive cells in the GCL rose significantly after NC. Seven days after NC, the number of RGCs in the CoPP-treated mice was significantly higher than in the control mice. Combined pretreatment with SnPP, an HO-1 inhibitor, suppressed the neuroprotective effect of CoPP. These results reflect changes in HO-1 activity to RGCs that are a key part of RGC survival. Upregulation of HO-1 signaling may therefore be a novel therapeutic strategy for glaucoma. (C) 2014 Wiley Periodicals, Inc.