VISUALIZATION AND QUANTIFICATION OF TRANSMURAL CONCENTRATION PROFILES OF MACROMOLECULES ACROSS THE ARTERIAL-WALL
VISUALIZATION AND QUANTIFICATION OF TRANSMURAL CONCENTRATION PROFILES OF MACROMOLECULES ACROSS THE ARTERIAL-WALL
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DOI：
10.1161/01.res.67.1.11
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发表时间：
1990-07-01
影响因子：
20.1
通讯作者：
CHISOLM, GM
中科院分区：
文献类型：
--
作者：
PENN, MS;KOELLE, MR;CHISOLM, GM
描述从血浆进入动脉壁的大分子的运输参数可以从进入组织的标记大分子的浓度谱中获得。一种测量这种浓度曲线的新技术已经开发出来，它提供的空间分辨率优于通过平行于内皮的平面连续切片组织来测量放射性标记大分子曲线的方法。此外，这种新方法保留了细胞组织和组织结构，并允许测量局灶性内皮损伤或血管病变的浓度曲线。该技术通过测量相关过氧化物酶活性来定量蛋白质的浓度。虽然本研究是使用辣根过氧化物酶（HRP）进行的，但同样的原理可以应用于与HRP或微过氧化物酶相关的其他大分子。利用图像处理系统对主动脉横切面HRP的彩色反应产物进行检测。在本研究中，通过与用放射标记HRP连续切片技术获得的大鼠主动脉中HRP浓度谱进行比较，验证了这种新方法获得的图谱。我们使用该技术测量正常动物内膜和介质中HRP的高分辨率浓度谱。这些浓度曲线表明，内部弹性层是大分子通过正常大鼠主动脉壁运输的主要屏障。这种新方法应该可以量化微循环血管周围组织、大血管增厚内膜内和冠状动脉壁内的大分子浓度谱。
Transport parameters that describe a macromolecule entering the arterial wall from plasma can be obtained from concentration profiles of the labeled macromolecule entering the tissue. A new technique has been developed for measuring such concentration profiles, which offers spatial resolution superior to methods that measure profiles of radiolabeled macromolecules by serially sectioning tissue in planes parallel to the endothelium. In addition, this new method preserves cellular organization and tissue structure and permits measurement of concentration profiles underlying focal endothelial injuries or vascular lesions. The technique quantifies the concentration of a protein by measuring associated peroxidase activity. Although the present study was performed using horseradish peroxidase (HRP), the same principles can be applied to other macromolecules linked to HRP or microperoxidase. The colored reaction product of HRP was detected in transverse aortic sections using an image processing system. In the present study, profiles obtained by this new method were validated by comparison with HRP concentration profiles in rat aortas obtained by a serial slicing technique using radiolabeled HRP. We used the technique to measure high-resolution HRP concentration profiles in the intima and media of normal animals. These concentration profiles suggest that the internal elastic lamina acts as a major barrier to transport of macromolecules across the wall of the normal rat aorta. The new method should allow concentration profiles for macromolecules to be quantified in tissue surrounding vessels in the microcirculation, within the thickened intima of large vessels, and across coronary artery walls.