Cellular aspects of vascular remodeling in hypertension revealed by confocal microscopy
Cellular aspects of vascular remodeling in hypertension revealed by confocal microscopy
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DOI：
10.1161/01.hyp.30.6.1455
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发表时间：
1997-12-01
期刊：
影响因子：
8.3
通讯作者：
McGrath, JC
中科院分区：
文献类型：
--
作者：
Arribas, SM;Hillier, C;McGrath, JC
在完整的动脉重塑的细胞方面还没有得到充分的研究，主要是由于缺乏一个适当的方法，允许简单的测量。本研究的目的是建立一种基于激光扫描共聚焦显微镜（LSCM）的方法，并与以往的方法进行比较，将其应用于高血压重构的研究。采用标准化直径设置（0.9（d100））的钢丝肌电描记术和计算的等效压力下的灌注肌电描记术，对易卒中型自发性高血压大鼠（SHRSP）和Wistar-Kyoto大鼠（WKY）的肠系膜阻力动脉的一段进行形态学测定。第二段用核染料Hoechst 33342（活组织）或碘化丙啶（固定组织）染色，并用LSCM和MetaMorph软件测量。与导丝肌造影相比，灌注肌造影显示与以前报道的相似的差异。与LSCM相比，灌注肌造影显示了相似的管腔，但在活组织和固定组织中的壁厚显著更小，可能是由于测量低估。在LSCM的研究中，SHRSP的动脉与WKY的动脉相比显示：（1）管腔减少;（2）细胞密度改变，外膜显著增加，中膜减少，内膜不变;（3）中膜体积显著增加;（4）内皮细胞核显著减小;（5）中膜中的外膜样细胞。我们的结论是：（1）LSCM是一种可靠和直接的方法来研究完整血管的形态学，（2）它提供了新的信息，重塑中的细胞变化，（3）外膜可能在高血压的重塑过程中发挥积极作用，（4）高血压的内皮“重塑”。
Cellular aspects of remodeling in intact arteries have not been fully investigated, mainly due to the lack of an appropriate methodology that allows for simple measurements. The aim of this study was to develop a method based on laser scanning confocal microscopy (LSCM), compare it with previous methodology, and apply it to the study of remodeling in hypertension. The morphology of mesenteric resistance arteries from stroke-prone spontaneously hypertensive rats (SHRSP) and Wistar-Kyoto rats (WKY) was determined with wire myography on one segment with a standardized diameter setting (0.9(d100)) and with perfusion myography on a second segment from the same artery at the calculated equivalent pressure. The second segments were stained with the nuclear dye Hoechst 33342 (live tissue) or propidium iodide (fixed tissue) and measured with LSCM and MetaMorph software. Compared with wire myography, perfusion myography showed similar differences from those previously reported. Compared with LSCM, perfusion myography showed a similar lumen but significantly smaller wall thickness in both live and fixed tissue, probably due to measurement underestimation. In the study with LSCM, arteries from SHRSP compared with those from WKY showed (1) reduced lumen, (2) altered cell density that was significantly increased in the adventitia, decreased in the media, and unchanged in the intima, (3) significantly increased medial volume, (4) significantly smaller endothelial cell nuclei, and (5) adventitial-like cells in the media. We conclude that (1) LSCM is a reliable and straightforward method to study morphology in intact vessels, (2) it provides new information on the cellular changes in remodeling, (3) adventitia might play an active role in the process of remodeling in hypertension, and (4) endothelium "remodels" in hypertension.