基于组织芯片技术探讨GLUT10在主动脉夹层发病机制中的作用

Study on the Role of GLUT10 Expression in the Pathogenesis of Aortic Dissection Based on Tissue Microarray

目的研究主动脉夹层患者的主动脉组织中葡萄糖转运体10(glucose transporter 10,GLUT10)表达水平及意义。方法选取2018年1~11月于武汉大学人民医院确诊为主动脉夹层的27例患者为夹层组,另选取18例心脏移植患者作为对照组。将以上患者的主动脉标本制作成组织芯片,检测GLUT10在人体主动脉组织中的蛋白表达水平及其相关性;采用载shRNA慢病毒敲低血管平滑肌细胞GLUT10编码基因,检测其周期蛋白、基质金属蛋白酶、细胞骨架蛋白的表达,探讨GLUT10表达下调致主动脉重构的病理机制。结果在组织芯片的免疫组化染色中发现,夹层组GLUT10的表达水平较对照组正常主动脉明显下调;提取GLUT10敲低后的细胞蛋白进行Western blot法检测发现,GLUT10敲低组的细胞周期蛋白、基质金属蛋白酶的表达水平较对照组明显增高,而细胞骨架蛋白的表达水平明显下降。结论GLUT10表达下调致血管平滑肌细胞周期紊乱、金属蛋白酶异常表达、细胞骨架蛋白表达下调、主动脉重构,与主动脉夹层的发生、发展密切相关。

Objective To investigate the expression and significance of glucose transporter 10(GLUT10)in aortic tissue of aortic dissection patients.Methods Twenty-seven patients diagnosed with aortic dissection in Renmin Hospital of Wuhan University from January 2018 to November 2018 were selected as the aortic dissection group,and another 18 patients receiving heart transplantation were selected as the control group.The aortic specimens of the above patients were made into tissue microarray,and the protein expression level of GLUT10 in human aortic tissues and its correlation were detected;shRNA was used to knock down GLUT10 coding gene in vascular smooth muscle cells,and the expression of cyclin,matrix metalloproteinase and cytoskeleton protein was detected to explore the pathological mechanism of aortic remodeling caused by GLUT10 down-regulation.Results The results of immunofluorescence staining of tissue-chip showed that GLUT10 expression in the aortic dissection group was significantly down-regulated compared with that in the normal aorta of the control group;Western blot experiment analysis of smooth muscle cells with GLUT10 lentivirus knockdown showed that the expression level of Cyclin and matrix metalloproteinases in GLUT10 knockdown group was significantly higher than that in control group,while the expression level of cytoskeleton protein was significantly decreased.Conclusion The down-regulated of GLUT10 expression leads to cell cycle disorder of vascular smooth muscle cells,abnormal expression of metalloproteinases,decreased expression of cytoskeletal proteins,and aortic remodeling,which is closely related to the occurrence and development of aortic dissection.