目的应用Realtime聚合酶链反应(PCR)、免疫组织化学技术观察OPN在夹层血管标本和正常对照标本中的表达是否存在差异。方法选取急性期的主动脉夹层标本(AD)标本12例,正常胸主动脉12例。抽提标本总RNA,以GAPDH基因为参照,采用RealtimePCR比较AD组和对照组OPN基因在RNA水平的差异;采用免疫组织化学方法显示OPN蛋白在两组标本中的表达,两组各个标本存低倍视野下随机选择3个部位进行阳性细胞计数,比较两组OPN在蛋白质水平表达的差异。结果AD组OPN基因的表达水平是正常组的5.56倍(P〈0.01),AD组OPN蛋白的表达水平是对照组的3.03倍(P〈0.01)。结论AD组标本OPN在基因和蛋白质水平均发生上调表达;OPN的上调表达可能与AD的形成密切相关。
Objective To identify the differential expression of osteopontin (OPN) between the aorta specimens from the aortic dissection (AD) and the normal controls. Methods Twelve descending aorta specimens from patients with acute Stanford B dissection were taken during surgical operation, and 12 normal descending aorta specimens were taken from multi-organ donors. The GAPDH gene was used as control, and mRNA and protein expression of OPN was detected by using real-time polymerase chain reaction (PCR) and immunohistochemisty, respectively. Three low-powered fields of each specimen in both groups were randomly chosen to count the cells positive for OPN protein. Results The expression levels of OPN in AD group was 5.56 folds as those in the control group (P 〈0. 01 ). The protein expression levels of OPN was 3.03 folds as those in the control group ( P 〈 0. 01 ). Conclusion The gene and protein expression levels of OPN were down-regulated in AD group as compared with normal controls. OPN is one of the markers to identify smooth muscle cells as synthetic phenotype. The up-regulation of OPN may contribute to the development of AD.