脏器微血管对荧光素钠通透性的实验方法

AN EXPERIMENTAL METHOD FOR STUDYING ORGAN MICROVASCULAR PERMEABILITY TO FLUORESCEIN SODIUM

大鼠颈动脉注射１％ＦｌＮａ，荧光显微镜下活体观察肠系膜微血管血流状态及ＦｌＮａ的渗出情况，并在不同时间点经股动脉采血，测定血浆内ＦｌＮａ浓度随时间的变化，利用组织匀浆测定不同脏器中ＦｌＮａ的分布，再辅以冰冻切片进行观察。活体观察发现，ＦｌＮａ注入体内后，经微血管迅速向周围组织渗出，最后汇集于淋巴管，血浆及组织匀浆ＦｌＮａ浓度的测定表明，ＦｌＮａ浓度随时间的变化呈指数衰减，各脏器ＦｌＮａ的分布极不相同。冰冻切片也显示了同样的分布差别。这些结果表明，我们所建立的方法可直观、定量地反映ＦｌＮａ在微血管的通透情况。

A calibration procedure was established to determine the microvascular and tissue concentration of fluorescein sodium (FINa) in the mesentery of the rat. After the tracer of 1% FINa was injected, selected venule and microvessels (diameter 10 30μm) were observed and videotaped by means of intravital fluorescence microscopy with a low illumination CCD camera. Simultaneously, arterial blood samples were taken to measure the changes of vascular FlNa concentration along with the time using a spectrofluormeter, a linear correlation between FlNa concentration and fluorescence intensity was obtained. Finally, different organs were dissected, homogenized and frozen sections were made. The interstitial FlNa concentration was measured with a spectrofluormeter. All those results demonstrated that the fluorescence intensity in each organ was different, indicating that our method is useful for studying the FlNa permeability of different organ microvessels.