Upregulation of chemokine expression in the retinal vasculature in ischemia-reperfusion injury
Upregulation of chemokine expression in the retinal vasculature in ischemia-reperfusion injury
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DOI：
10.1167/iovs.02-1308
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发表时间：
2003-09-01
影响因子：
4.4
通讯作者：
Rao, NA
中科院分区：
文献类型：
--
作者：
Jo, N;Wu, GS;Rao, NA
目的.为了评估缺血再灌注损伤后不同视网膜部位的趋化因子表达，采用激光捕获显微切割获得的选定组织进行逆转录-聚合酶链反应（RT-PCR）分析。在刘易斯大鼠中通过增加眼内压75分钟来产生视网膜缺血。在再灌注后3、6、12和24小时，采用RT-PCR法检测单核细胞趋化蛋白（MCP）-1、巨噬细胞炎性蛋白（MIP）-1 α、MIP-1 β、白细胞介素（IL）-8和干扰素-γ诱导的10-kDa蛋白（IP-10）mRNA在神经节细胞层（GCL）、内核层（INL）、外核层（ONL）、和视网膜血管，在这些视网膜层的激光捕获显微切割之后。这些趋化因子通过免疫组织化学方法，使用MCP-1和MIP-1 α特异性抗体进一步定位。白细胞共同抗原免疫组化染色检测视网膜内白细胞浸润。缺血再灌注诱导MCP-1、MIP-1 α和MIP-1 β mRNA在再灌注后3小时视网膜血管中表达。再灌注6小时后，GCL和INL中可见这些趋化因子和IL-8 mRNA的表达。再灌注12 h后，IP-10 mRNA在GCL和INL中表达。再灌注后24小时，在GCL、INL和视网膜血管中检测到免疫反应性MCP-1和MIP-1 α。在任何时间ONL中均未检测到趋化因子mRNA表达或免疫反应性。白细胞浸润在12 h时可见，再灌注后24 h明显增加。缺血-再灌注视网膜损伤导致高度趋化剂的产生，最初在视网膜脉管系统中，然后在其他视网膜内层中。这种差异性趋化因子表达可能在白细胞募集和选择性白细胞浸润内层视网膜中起作用，导致主要局限于神经节细胞和其他内层神经元结构的视网膜损伤。
Purpose. To evaluate chemokine expression at various retinal sites after ischemia-reperfusion injury, using reverse transcription-polymerase chain reaction (RT-PCR) analysis of selected tissue obtained by laser capture microdissection.Methods. Retinal ischemia was produced in Lewis rats by increasing intraocular pressure for 75 minutes. At 3, 6, 12, and 24 hours after reperfusion, RT-PCR was used to measure the levels of monocyte chemoattractant protein (MCP)-1, macrophage inflammatory protein (MIP)-1alpha, MIP-1beta, interleukin (IL)-8, and interferon-gamma-inducible 10-kDa protein (IP-10) mRNA expression in the ganglion cell layer (GCL), inner nuclear layer (INL), outer nuclear layer (ONL), and retinal vessels, after laser capture microdissection of these retinal layers. These chemokines were further localized by immunohistochemical methods, using antibodies specific to MCP-1 and MIP-1alpha. Leukocyte infiltration into the retina was detected with immunostaining for leukocyte common antigen.Results. Ischemia-reperfusion induced expression of MCP-1, MIP-1alpha, and MIP-1beta mRNA in the retinal vessels 3 hours after reperfusion. Six hours after reperfusion, expression of these chemokines and IL-8 mRNA was seen in the GCL and INL. Twelve hours after reperfusion, IP-10 mRNA expression was seen in the GCL and INL. Immunoreactive MCP-1 and MIP-1alpha were detected in the GCL, INL, and the retinal vessels 24 hours after reperfusion. No chemokine mRNA expression or immunoreactivity was detected in the ONL at any time. Leukocyte infiltration was noted at 12 hours, increasing markedly 24 hours after reperfusion.Conclusions. Ischemia-reperfusion retinal injury results in generation of highly chemotactic agents, initially in the retinal vasculature, then in the other inner retinal layers. Such differential chemokine expression may play a role in leukocyte recruitment and selective leukocyte infiltration in the inner retina, leading to retinal damage primarily localized to the ganglion cells and other inner neuronal structures.