Intestinal ischemia-reperfusion injury: reversible and irreversible damage imaged in vivo
Intestinal ischemia-reperfusion injury: reversible and irreversible damage imaged in vivo
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DOI：
10.1152/ajpgi.90595.2008
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发表时间：
2009-07-01
影响因子：
4.5
通讯作者：
Montrose, Marshall H.
中科院分区：
文献类型：
--
作者：
Guan, Yanfang;Worrell, Roger T.;Montrose, Marshall H.
Guan Y，Worrell RT，Pritts TA，蒙特罗斯MH.肠缺血再灌注损伤：可逆性和不可逆性损伤的体内成像。美国生理学杂志胃肠和肝脏生理学297：G187-G196，2009年。首次发表于2009年4月30日; doi：10.1152/ajpgi.90595.2008。肠缺血发作的早期事件难以评估。使用体内显微镜，我们已经实时分析了短期（15分钟）和长期（40-50分钟）缺血与随后的再灌注（IR）的影响，评价结构，完整性和功能的小鼠空肠粘膜，同时通过共聚焦显微镜监测血流。通过血管封堵器的充气/放气施加IR，监测血流并通过扫描共聚焦成像确认。短暂缺血后，绒毛尖端细胞内NAD（P）H浓度迅速升高（23%）（共聚焦自体荧光显微镜），pH敏感探针BCECF显示细胞内pH（pH（i））的双相响应，从静止值6.8 +/- 0.1快速碱化至7.1 +/- 0.1，但随后强烈酸化至6.3 +/- 0.1。再灌注后，数值恢复至对照组。与此相反，结果是异质性后，长IR。在长时间缺血，三分之一的上皮细胞保持活力与可逆的变化，再灌注后，但剩余的细胞失去了膜的完整性（荧光黄吸收，LY），并在缺血过程中膜泡。随着再灌注间隔的进展，当细胞表现出更严重的NAD（P）H和pHi值、更大的水泡和更多的LY摄取并最终从绒毛脱落时，这些作用变得更加明显。从立体显微镜的结果表明，这些不可逆的影响，IR可能已经发生的局部血流，特别是在肠的抗肠系膜侧的不完全澄清的结果。我们的结论是短暂缺血对空肠上皮细胞的不良影响是完全可逆的再灌注间隔期间。然而，在长时间缺血后，再灌注不能恢复大多数细胞的正常结构和功能，这些细胞进一步恶化。我们的研究结果提供了一个基础，定义细胞的事件，导致组织从可逆的过渡到不可逆的损害在IR。
Guan Y, Worrell RT, Pritts TA, Montrose MH. Intestinal ischemia-reperfusion injury: reversible and irreversible damage imaged in vivo. Am J Physiol Gastrointest Liver Physiol 297: G187-G196, 2009. First published April 30, 2009; doi:10.1152/ajpgi.90595.2008.-The early events in an intestinal ischemic episode have been difficult to evaluate. Using in vivo microscopy we have analyzed in real-time the effects of short (15 min) and long (40-50 min) ischemia with subsequent reperfusion (IR), evaluating structure, integrity, and functioning of the mouse jejunal mucosa while monitoring blood flow by confocal microscopy. IR was imposed by inflation/deflation of a vascular occluder, and blood flow was monitored and confirmed with scanning confocal imaging. After short ischemia, villus tip cells revealed a rapid increase (23%) in the intracellular NAD(P)H concentration (confocal autofluorescence microscopy), and the pH-sensitive probe BCECF showed a biphasic response of the intracellular pH (pH(i)), quickly alkalinizing from the resting value of 6.8 +/- 0.1 to 7.1 +/- 0.1 but then strongly acidifying to 6.3 +/- 0.1. Upon reperfusion, values returned toward control. In contrast, results were heterogeneous after long IR. During long ischemia, one-third of the epithelial cells remained viable with reversible changes upon reperfusion, but remaining cells lost membrane integrity (Lucifer Yellow uptake, LY) and had membrane blebs during ischemia. These effects became more pronounced as the reperfusion interval progressed when cells exhibited more severely affected NAD(P) H and pHi values, larger blebs, and more LY uptake and eventually were shed from the villus. Results from stereo microscopy suggest that these irreversible effects of IR may have occurred as a result of incomplete restorations of local blood flow, especially at the antimesenteric side of the intestine. We conclude that the adverse effects of short ischemia on the jejunum epithelium are fully reversible during the reperfusion interval. However, after long ischemia, reperfusion cannot restore normal structure and functioning of a majority of cells, which deteriorate further. Our results provide a basis for defining the cellular events that cause tissue to transit from reversible to irreversible damage during IR.