Evans blue staining of cardiomyocytes induced by myocardial contrast echocardiography in rats: Evidence for necrosis instead of apoptosis
Evans blue staining of cardiomyocytes induced by myocardial contrast echocardiography in rats: Evidence for necrosis instead of apoptosis
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DOI：
10.1016/j.ultrasmedbio.2007.06.008
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发表时间：
2007-12-01
影响因子：
2.9
通讯作者：
Gordon, David
中科院分区：
文献类型：
--
作者：
Miller, Douglas L.;Li, Peng;Gordon, David
具有对比剂的高机械指数（MI）超声心动图已被证明会诱导心肌细胞的埃文斯蓝色染色，在暴露后1 d（除了收缩带坏死），在暴露后立即看到。这项研究检查了坏死与凋亡在这些生物效应中的作用。高MI处的心肌对比度超声心动图在1：4的心电图触发下以1.5 MHz进行麻醉大鼠进行。通过在扫描开始10分钟开始时，通过尾静脉注入高剂量的50 mU L/kg超声对比度，累积了50 mu/kg超声对比介质，从组织学上观察到的细胞损伤。将埃文斯蓝色染料或碘化丙啶注入为心肌细胞质膜完整性的指标。使用末端DUTP Nick-End标记（TUNEL）方法染色的组织学切片，用于将核与DNA降解（例如凋亡）染色。将埃文斯蓝色荧光细胞计算在冷冻切片或苏木精染色和tunel标记的石蜡切片上。此外，使用透射电子显微镜评估潜在的凋亡核。成像暴露后立即观察到过度收获和碘化丙啶染色。尽管在4小时后明显显示了TUNEL阳性细胞，但这些细胞也有收缩带坏死的迹象，并且没有通过电子显微镜证实凋亡的特征。 24小时后，炎症细胞浸润是明显的。埃文斯蓝色染色认识到第二次，更微妙的损伤，在24小时后，在形态上完整的染色细胞处的炎性细胞浸润最小。在24小时内用埃文斯蓝色染色的心肌细胞中的TUNEL方法未检测到凋亡。然而，埃文斯蓝染的细胞数量在48小时后下降，并持续炎症细胞浸润。埃文斯蓝染色的心肌细胞的最初侮辱显然引起了质膜的部分渗透性，这导致了24至48小时后逐渐变性（但不是凋亡）和坏死。 （电子邮件：douglm@umich.edu）（c）2007年世界医学与生物学超声联合会。
High mechanical index (MI) echocardiography with contrast agent has been shown to induce Evans blue staining of cardiomyocytes, seen 1 d after exposure, in addition to contraction band necrosis, seen immediately after exposure. This research examined the roles of necrosis vs. apoptosis in these bioeffects. Myocardial contrast echocardiography at high MI with 1:4 electrocardiogram triggering was performed in anesthetized rats at 1.5 MHz. Histologically observable cell injury was accumulated by infusing a high dose of 50 mu L/kg ultrasound contrast media via tail vein for 5 min at the start of 10 min of scanning. Evans blue dye or propidium iodide was injected as an indicator of cardiomyocyte plasma membrane integrity. Histologic sections were stained using the terminal dUTP nick-end labeling (TUNEL) method for labeling nuclei with DNA degradation (e.g., apoptosis). Evans blue fluorescent cells were counted on frozen sections or on hematoxylin-stained and TUNEL-labeled paraffin sections. In addition, transmission electron microscopy was used to assess potential apoptotic nuclei. Hypercontraction and propidium iodide staining were observed immediately after imaging exposure. Although TUNEL-positive cells were evident after 4 h, these also had indications of contraction band necrosis, and features of apoptosis were not confirmed by electron microscopy. Inflammatory cell infiltration was evident after 24 h. A second, more subtle injury was recognized by Evans blue staining, with minimal inflammatory cell infiltration at the morphologically intact stained cells after 24 h. Apoptosis was not detected by the TUNEL method in the cardiomyocytes stained with Evans blue at 24 h. However, Evans blue-stained cell numbers declined after 48 h, with continued inflammatory cell infiltration. The initial insult for Evans blue-stained cardiomyocytes apparently induced partial permeability of the plasma membrane, which led to gradual degeneration (but not apoptosis) and necrosis after 24 to 48 h. (E-mail: douglm@umich.edu) (c) 2007 World Federation for Ultrasound in Medicine & Biology.