Effects of Ischemic Preconditioning and Bevacizumab on Apoptosis and Vascular Permeability Following Retinal Ischemia-Reperfusion Injury
Effects of Ischemic Preconditioning and Bevacizumab on Apoptosis and Vascular Permeability Following Retinal Ischemia-Reperfusion Injury
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DOI：
10.1167/iovs.10-5264
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发表时间：
2010-11-01
影响因子：
4.4
通讯作者：
Antonetti, David A.
中科院分区：
文献类型：
--
作者：
Abcouwer, Steven F.;Lin, Cheng-mao;Antonetti, David A.
目的。本研究采用短暂性缺血再灌注（IR）模型，比较缺血预处理（IPC）和靶向血管内皮生长因子（VEGF）和肿瘤坏死因子(TNF)- α治疗对视网膜凋亡、血管通透性和mRNA表达的影响。大鼠进行30或45分钟的视网膜缺血，然后再灌注长达48小时。用caspase-3 （DEVDase）活性和核小体DNA含量（细胞死亡ELISA）测定神经退行性变。用Evans蓝染色法定量血管渗漏。通过全基因组芯片鉴定出一组ir应答mrna，并通过RT-PCR分析证实。Western blot法检测VEGF蛋白含量。IPC在IR前24小时缺血10分钟完成。玻璃体内注射贝伐单抗或依那西普可抑制VEGF和TNF α信号。IR作用4和48 h后，视网膜细胞凋亡和血管通透性显著升高。IR降低了VEGF mRNA，但VEGF蛋白显著升高。IPC有效抑制神经变性，贝伐单抗有效抑制血管通透性，依那西普没有影响这两种结果。IPC显著改变了33个IR反应性mrna中的15个的IR反应，而贝伐单抗对这些mrna没有显著影响。IR提供了一种急性缺血性视网膜病变模型，包括神经变性和vegf依赖性血管通透性，并且适合快速药物治疗测试。IPC和贝伐单抗的不同作用表明，IR的细胞凋亡和血管反应可能是分开的，针对每个病理终点的治疗方法可能是治疗缺血性视网膜疾病的必要条件。（Invest Ophthalmol Vis Sci. 2010;51:59 920-5933） DOI:10.1167/iovs.10-5264
PURPOSE. Using transient ischemia followed by reperfusion (IR) to model ischemic retinal disease, this study compares the effects of ischemic preconditioning (IPC) and therapies targeting vascular endothelial growth factor (VEGF) and tumor necrosis factor (TNF)-alpha on retinal apoptosis, vascular permeability, and mRNA expression.METHODS. Rats were subjected to 30 or 45 minutes of retinal ischemia followed by reperfusion for up to 48 hours. Neurodegeneration was quantified by caspase-3 (DEVDase) activity and by measuring nucleosomal DNA content (cell death ELISA). Vascular leakage was quantified by the Evans Blue dye method. A set of IR-responsive mRNAs was identified by whole-genome microarray and confirmed by RT-PCR analyses. VEGF protein was measured by Western blot analysis. IPC was accomplished with 10 minutes of ischemia 24 hours before IR. VEGF and TNF alpha signaling was inhibited by intravitreal injection of bevacizumab or etanercept, respectively.RESULTS. IR caused significant retinal cell apoptosis and vascular permeability after 4 and 48 hours. Whereas IR decreased VegfA mRNA, VEGF protein was significantly increased. IPC effectively inhibited neurodegeneration, bevacizumab effectively inhibited vascular permeability, and etanercept failed to affect either outcome. IPC significantly altered the IR responses of 15 of 33 IR-responsive mRNAs, whereas bevacizumab had no significant effect on these mRNAs.CONCLUSIONS. IR provides an acute model of ischemic retinopathy that includes neurodegeneration and VEGF-dependent vascular permeability and is amenable to rapid drug therapy testing. The distinct effects of IPC and bevacizumab demonstrate that the apoptotic and vascular responses to IR may be separated and that therapeutics targeting each pathologic endpoint may be warranted in treating ischemic retinal diseases. (Invest Ophthalmol Vis Sci. 2010;51:5920-5933) DOI:10.1167/iovs.10-5264