己糖-6-磷酸脱氢酶在急性主动脉夹层患者主动脉壁组织中表达变化的研究

Expression and implication of H6PD in the aortic wall of patients with acute aortic dissection

目的:探究急性主动脉夹层患者主动脉壁组织中己糖-6-磷酸脱氢酶(H6PD)的表达变化及其意义。方法:纳入2015年至2019年收治于我科并行手术治疗的59例急性Stanford A型主动脉夹层患者作为夹层组,选取同时段内接受心脏移植手术的22例移植受体作为对照组,手术过程中剪取部分主动脉壁标本。采用苏木素-伊红(HE)染色及弹性纤维(EVG)染色观察两组患者主动脉壁组织结构的差异;提取主动脉组织蛋白,蛋白质印迹法(Western blot)检测主动脉壁组织中H6PD蛋白表达,应用Imagelab软件进行蛋白定量;免疫组化染色法检测两组样本中H6PD蛋白表达以及亚细胞定位,并使用Image J软件对阳性染色面积进行定量;使用GraphPad Prism 8.3.0软件进行组间差异的比较。结果:HE与EVG染色显示夹层组主动脉壁组织中平滑肌细胞丢失且排列紊乱,伴有弹力纤维的减少和断裂;Western blot结果显示H6PD蛋白在夹层组主动脉壁组织中表达上调;免疫组化染色显示H6PD分子定位于胞质,与对照组比较,夹层组主动脉壁组织中H6PD蛋白表达水平显著升高。结论:主动脉夹层患者主动脉壁组织中H6PD蛋白表达升高,提示该分子可能参与主动脉夹层的发生发展。

Objective:To investigate the expression changes and clinical significance of hexose-6-phosphate dehydrogenase(H6PD)in the aortic wall of patients with acute aortic dissection.Methods:From 2015 to 2019,59 patients who underwent concurrent surgery for Stanford type A aortic dissection in our division were included as the aortic dissection(AD)group,22 transplant recipients who underwent heart transplantation during the same period were selected as the control group.We performed Hematoxylin-eosin(HE)and Verhoeff's Van Gieson(EVG)staining to observe the difference in microstructure of the aortic wall tissue between the two groups;the expression level of H6PD protein in aorta tissue was estimated through Western blot,and protein quantification was conducted using Image lab software;immunohistochemical staining was adopted to confirm the distinction of H6PD protein expression and show the subcellular localization of H6PD,the positive staining area was quantified utilizing Image J software;GraphPad Prism 8.3.0 software was used for comparison of inter-group differences.Results:In AD group,HE staining presented that the smooth muscle cells were lost and disarranged in aortic wall,corresponding degradation and breakage of elastic fibers was observed in AD medial aorta through EVG staining;the results of Western blot demonstrated an up-regulated protein expression of H6PD in AD aorta tissue compared with the control group;the results of immunohistochemical staining showed that H6PD was mainly located in cytoplasm and further verified the elevated protein level of H6PD in AD group.Conclusion:The upregulation of H6PD in AD patients indicates that H6PD may participate in the process of AD development.