Analysis of microcirculation of experimental metastatic lung tumor using artificial oxygen carrier as a tool for altering the microvascular tone

肺内转移瘤的供氧环境和微循环环境与其他器官的转移瘤相比有很大的不同。为了研究转移性肺肿瘤微循环的建立，必须建立肺转移的同源模型。为了观察肺微循环，我们在大鼠胸壁植入了一个水晶窗腔。通过这个窗口，我们可以用激光共聚焦显微镜和荧光染料观察肺小动脉、肺泡毛细血管和肺静脉。将图像记录在VCR中，然后进行分析。可以测量血管内径，评价毛细血管区域的血流情况。测量肺小动脉的血流需要更复杂的技术，因为血流速度足够快，可以捕获单个荧光标记的红细胞。为了建立肺转移模型，我们用LY80建立了冬雨大鼠腹水炎性肝癌模型。已检查的modaliti…更多的ES是通过腹腔注射肿瘤细胞、皮下种植肿瘤细胞、开胸直接将肿瘤细胞移植到肺内、尾静脉输注肿瘤细胞、尾静脉输注皮下移植瘤小组织悬液。在这些方法中，尾静脉输注皮下种植肿瘤的小组织悬浮液给我们提供了最可靠和稳定的结果。在注射后15~17天，我们可以评估肺转移瘤。为了评估肿瘤微血管的反应性，我们使用了氧合血红蛋白溶液和乙肝病毒悬液。氧合血红蛋白溶液具有很强的收缩微血管的潜力，而最初作为人工氧载体发展起来的乙肝病毒被认为对微血管几乎没有影响。在正常动物中，肺小动脉在注射氧合血红蛋白后收缩，而在注射乙肝病毒后观察到一过性收缩。肺泡毛细血管直径、流量变化不大。然而，肺小动脉对这些挑战的反应揭示了一氧化氮是决定肺血管张力的主要调节剂之一。在肿瘤负荷动物中，特别是在当前的模型中，肺泡毛细血管扩张。但未观察到微动脉和/或小静脉面积的变化。在这个模型中，氧合血红蛋白和乙肝病毒的作用与在完整动物中观察到的相似。我们必须改进模型来分析转移性肺肿瘤微循环中的事件。较少

The environment of metastatic tumor in the lung is much different in terms of oxygen supply and microcirculation compare to those metastatic tumors in other organs. To investigate the establishment of microcirculation of metastatic lung tumor, we have to establish syngenic model of lung metastasis.To observe lung microcirculation, we implanted a crystal window chamber in the chest wall of a rat. Through this window, we could observe pulmonary arteriole, alveolar capillary, and pulmonary vein using laser confocal microscope and fluorescent dye. The images were recorded in VCR and analyzed afterwards. Inner diameter of vasculature could be measured and blood flow could be evaluated in capillary area. More sophistication was necessary to measure the blood flow in pulmonary arteriole because flow was fast enough to capture individual fluorescent labeled red blood cells. To establish lung metastasis, we evaluated 5 models using LY80 which is ascitis hepatoma in Donryu rat. Examined modaliti … More es were as follows, intraperitoneal tumor cell injection, subcutaneous implantation of tumor cells, implantation of tumor cells to the lung directly by thoracotomy, tail vein infusion of tumor cells, and tail vein infusion of small tissue suspension taken from subcutaneous implanted tumor. Among these modalities, tail vein infusion of small tissue suspension taken form subcutaneous implanted tumor gave us the most reliable and stable results. After 15 to 17 days after infusion, we could evaluate the pulmonary metastatic tumor.To evaluate the reactivity of tumor microvasculature, we used oxyhemoglobin solution and HbV suspension. Oxyhemoglobin solution is a strong potential of contraction of microvasculature, while HbV (liposome encapsulated hemoglobin), which is originally developed as an artificial oxygen carrier, is thought to have little effect on microvasculature. In normal animals, pulmonary arterioles contract after injection of Oxyhemoglobin while transient contraction was observed after HbV injection. There are little change of diameter, flow in alveolar capillary. However, pulmonary arteriole react these challenges revealing that Nitric Oxide is one of major modulator in determining the tone of pulmonary vasculature.In tumor burden animals, especially in current model, alveolar capillary was dilated. But we could not observe any changes in arteriole and/or venule area. The effect of oxyhemoglobin and HbV in this model is similar to those observed in intact animals. We have to improve the model to analyze the event in microcirculation of metastatic lung tumor. Less