主动脉夹层患者体外循环术后红细胞超微形态学观察

The ultrastructural morphological changes of red blood cells in aortic dissection patients undergoing extracorporeal circulation

目的应用显微影像技术观察主动脉夹层患者体外循环术后红细胞超微形态学变化,为红细胞损伤和亚损伤提供超微形态学证据。方法收集本中心2016年3月到2016年8月20例主动脉夹层患者手术前后的血液标本。使用不同显微技术(扫描电镜、原子力显微镜和激光共聚焦显微镜)观察红细胞超微形态学改变(红细胞畸形率、红细胞长度、宽度、高度、洞深、表面粗糙度等)以及红细胞膜的完整性(丝状激动蛋白分布)。结果与健康成人对照组相比,主动脉夹层患者的红细胞微观形态学参数除表面粗糙度外,其它微观形态学参数(高度、宽度、洞深、粗糙度)没有明显变化。在经历体外循环术后红细胞形态发生明显变化,部分红细胞出现肿胀、固缩、变形、破裂以及红细胞碎片,出现各种异常形态的红细胞(棘型、口型、靶型、椭圆形、薄型等)。红细胞膜表面光滑度改变,细胞表面出现高低起伏大小不等的凸起颗粒使细胞膜表面粗糙度增加,同时细胞表面丝状激动蛋白分布改变,细胞结构呈现解聚状态,不能形成完整的荧光环,红细胞膜结构的完整性受到破坏。结论超微形态学改变为夹层疾病和体外循环引起红细胞损伤提供了新的形态学证据。红细胞表面粗糙度可以作为新的参数来评估红细胞损伤以及亚损伤。

Objective This study aimed to observe the morphological changes of red blood cells（ RBCs） in aortic dissection（ AD） patients undergoing extracorporeal circulation（ ECC） by different microscopic technologies. Methods 20 cases were included in this study. The ultrastructural morphology of RBCs（ malformation percentage; length; width; height; concave depth; roughness; filamentous actin） was observed by scanning electron microscopy（ SEM）,laser scanning confocal microscope（ LSCM） and atomic force microscopy（ AFM）. Results Most of morphological parameters showed no obvious changes except roughness in AD patients compared with healthy controls. Almost all morphological parameters showed changes after ECC. Abnormal cells such as acanthocytes and echinocytes appeared and different sizes of protrusions also appeared on the cell membrane surface. Surface roughness of RBCs increased and the cell membrane skeleton was damaged. No complete red fluorescent ring formed anymore. Conclusion Our research offers some valuable information regarding RBCs damage in AD disease and ECC. It also provides morphological basis for the study of RBCs functional change. Cell membrane surface roughness could be identified as a new parameter for evaluating the degree of RBCs damage in AD patients and the biocompatibility of ECC device.