Institute of Clinical Medicine, Aarhus University, Aarhus, Denmark ; Department of Clinical Physiology and Nuclear Medicine, Aarhus University Hospital, Skejby, Denmark.
Institute of Clinical Medicine, Aarhus University, Aarhus, Denmark ; Department of Urology, Aarhus University Hospital, Skejby, Denmark.
EJNMMI Res. 2013 Aug 7;3:62. doi: 10.1186/2191-219X-3-62. eCollection 2013.
The purpose of the present study was to quantify renal cell injury after ischemia and reperfusion in a pig model using (99m)Tc-lactadherin as a marker of apoptosis and (99m)Tc-sestamibi as a marker of mitochondrial dysfunction.
Thirty-four pigs were randomized into unilateral renal warm ischemia of 120 (WI120) or 240 min (WI240). The glomerular filtration rate (GFR) was calculated by renal clearance of (51)Cr-ethylenediaminetetraacetic acid, and apoptosis was quantified by immunohistochemical detection of caspase-3. After 240 min of reperfusion, intravenous (99m)Tc-lactadherin or (99m)Tc-sestamibi was injected simultaneously with (153)Gd microspheres into the aorta. Ex-vivo static planar images of the kidneys were acquired for determination of the differential renal function of tracer distribution using a gamma camera.
In WI120, there was no significant difference in the uptake of microspheres in the ischemic and contralateral normal kidney indicating adequate perfusion (uptake in ischemic kidney relative to the sum of uptake in both kidneys; 46% ± 12% and 51% ± 5%). In WI240, the uptake of microspheres was severely reduced in both groups (17% ± 11% and 27% ± 17%). GFR was severely reduced in the post ischemic kidney in both groups. In both groups, the uptake of lactadherin was reduced (41% ± 8%, 17% ± 13%) but not different from the uptake of (153)Gd microspheres. Caspase-3-positive cell profiles were increased in the post-ischemic kidneys (p < 0.001) and increased as the length of ischemia increased (p = 0.003). In both WI120 and WI240, the amount of (99m)Tc-sestamibi in the ischemic kidney was significantly lower than the amount of (153)Gd microspheres (40 ± 5 versus 51 ± 5 and 20 ± 11 versus 27 ± 17; p < 0.05).
In an established pig model with unilateral renal warm ischemia, we found significantly reduced (99m)Tc-sestamibi uptake relative to perfusion in the kidneys exposed to ischemia indicating a potential ability to detect renal ischemic and reperfusion injuries. However, apoptosis was not detected using (99m)Tc-lactadherin in the post-ischemic kidneys despite increased number of caspase-3-positive cell profiles.
This study is approved by the Danish Inspectorate of Animal Experiments (2010/561-1837).
本研究旨在通过使用（99m）Tc-乳白蛋白作为细胞凋亡标志物和（99m）Tc-甲氧基异丁基异腈作为线粒体功能障碍标志物，定量评估猪模型缺血再灌注后肾细胞损伤。
34 头猪随机分为单侧肾热缺血 120 分钟（WI120）或 240 分钟（WI240）组。肾小球滤过率（GFR）通过肾脏对（51）Cr-乙二胺四乙酸的清除率计算，细胞凋亡通过 caspase-3 免疫组化检测进行定量。再灌注 240 分钟后，将（99m）Tc-乳白蛋白或（99m）Tc-甲氧基异丁基异腈同时静脉注射到主动脉中，同时注射（153）Gd 微球。使用伽马相机获得肾脏的离体静态平面图像，以确定示踪剂分布的差异肾功能。
在 WI120 中，缺血侧和对侧正常侧肾脏的微球摄取没有显著差异，表明灌注充足（缺血侧肾脏的摄取相对于双侧肾脏摄取的总和；46%±12%和 51%±5%）。在 WI240 中，两组的微球摄取均严重减少（17%±11%和 27%±17%）。两组缺血侧肾脏的 GFR 均严重降低。两组的乳白蛋白摄取均减少（41%±8%，17%±13%），但与（153）Gd 微球的摄取无差异。缺血后肾脏的 caspase-3 阳性细胞谱增加（p<0.001），并且随着缺血时间的延长而增加（p=0.003）。在 WI120 和 WI240 中，缺血肾脏中（99m）Tc-甲氧基异丁基异腈的量明显低于（153）Gd 微球的量（40±5 比 51±5 和 20±11 比 27±17；p<0.05）。
在单侧肾热缺血的已建立猪模型中，我们发现与缺血暴露的肾脏灌注相比，（99m）Tc-甲氧基异丁基异腈的摄取显著减少，表明其具有检测肾缺血再灌注损伤的潜力。然而，尽管 caspase-3 阳性细胞谱增加，但在缺血后肾脏中仍未检测到（99m）Tc-乳白蛋白的细胞凋亡。
本研究已获得丹麦动物实验监察署批准（2010/561-1837）。
